### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

1 Data Requirement: PMRA DATA CODE: 9.5.2.1; 9.5.2.2; 9.5.2.3; 9.5.2.4 EPA DP Barcode: Not applicable OECD Data Point: IIA 8.2.1 and IIA 8.2.1.2 EPA Guideline: 850.1075

Test material: Technical and Formulation Purity (%): TGAI 98% and EP 48% Common name : Fluridone Chemical name : 1-methyl-3-phenyl-5-( α , α , α -trifluoro- m -tolyl)-4-pyridone IUPAC : 1-methyl-3-phenyl-5-( α , α , α -trifluoro-m-tolyl)-4-pyridone CAS name : 1-methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4(1 H )-pyridinone CAS No. : 59756-60-4 Synonyms :

Primary Reviewer: Katrina White Date: February 24, 2010 USEPA/OPPTS/OPP/ERBII/Biologist

Secondary Reviewer(s): Donna M. Randall Date: February 24, 2010 USEPA/OPPTS/OPP/ERBII/Senior Effects Scientist

Company Code: [For PMRA] Active Code: [For PMRA] Use Site Category: [For PMRA] EPA PC Code: 112900

CITATION : U.S. Fish and Wildlife Service. (1982). The Toxicity of Fluridone (El-171, Compound 112371) and of Sonar AS, an Aqueous Suspension Form ulation of Fluridone, to Aquatic Invertebrates and Fish in Acute and Chronic Studies. (U npublished study received Jun 10, 1982 under 1471-126; prepared in cooperation with U.S. Fish and Wild life Service, Columbia National Fisheries Research Laboratory, submitted by Elanco Products Co., Di v. of Eli Lilly and Co., Indianapolis, IN; CDL:070932-F). MRID 000103297

The acute endpoints from this study are identical to those reported in: Mayer, F., and M. Ellersieck (M&E) (1986). Manual of Acute Toxicity: Interpretation and Data Base for 410 Chemicals and 66 Species of Freshwater Animals . US Fish & Wildlife Service, Resource Publication 160. 579 p. MRID 40098001

Raw data were obtained from the Fede ral Lab Data submitted to the EFED.

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 fish. 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 scientifi cally sound and whether the study satisfies any applicable data requirements. Reviewers are e xpected to review and to determine for each study, on a case-by-case basis,

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

2 whether it is scientifically sound and provides sufficient in formation 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 revi ewer is to take into account the totality of factors related to the test methodology and results in determining the acceptability of the study.

The following template illustrates the critical elements and the level of detail required by the PMRA and the EPA in a study review of the acute toxicity to fish. It is not meant to be a "fill in the blanks" exercise although it may be appropriate in some cases. The standard format of the template should, however, be followed in preparing the study review.



### EXECUTIVE SUMMARY :

In 96-hr acute toxicity tests, rainbow trout ( Oncorhynchus mykiss formerly Salmo gairdneri ), bluegill sunfish ( Lepomis macrochirus ), channel catfish ( Ictalurus punctatus ), and fathead minnnow (Pimephales promelas ) were exposed to technical grade active ingred ient (TGAI) and an end-use product (EP), Sonar 4 AS. Rainbow trout and channel catfish were e xposed at two hardness levels (40 and 320 mg/L as calcium carbonate,CaCO 3 

### ), three temperatures (7, 12, and 17 for rainbow trout and 7, 12, and 22 for channel catfish), and three pH (6.5, 7.5, and 8.5). Nominal concentrations in the tests varied but were typically 0 (negative control and solvent control (acet one) for TGAI studies), 1.8, 3.2, 5.6, 10, 18, and 32 mg a.i./L under static conditions, additional nominal c oncentrations tested included 0.56, 1.0, 56, and 100 mg a.i./L.

Except for one test, fluridone concentrations were measured twice in the tests (test initiation and termination); however, the measured concentrations were only summarized in the USFWS (1982) study report. In some tests, the average measured concentrations were reportedly ≥ 79% of nominal. In other tests, average concentrations were <79% of nomin al or samples were lost. In M&E (1986) and the USFWS (1982) study report results are based on nominal . However, use of nominal concentrations to represent soluble fluridone exposures in the case of no anal ytical confirmatory data or with conditions of poor agreement with nominal concentrations poten tially underestimates the toxicity of fluridone. Fluridone has a short degradation rate and all test s included concentrations of fluridone above its solubility; one study actually reported insoluble fluridone at concentrations used in all tests. Therefore, if no sample analysis was available, the reviewer ex cluded the treatments above solubility from calculating the LC 50 

### if mortality levels allowed, and all these results were identified as based on total fluridone. Even where there was indication of agreement with nomin al concentrations, the reviewer did not include concentrations above solubility if mortality was high in concentrations below solubility, in calculating the LC 50 

### values. If mortality occurred primarily above so lubility, the results were annotated as total because there is a high degree of uncertainty of soluble flur idone concentrations without looking at the specific fluridone treatment concentrations.

The lowest quantitative TGAI 96-h LC 50 

### for rainbow trout was 5.1 mg a.i./L (mean measured). The lowest quantitative TGAI 96-hr LC 50 

### for channel catfish was 7.5 mg a.i./L (based on nominal concentration, however anlyatical analysis indicates that nominal concentrations were reliable). An endpoint could be calculated for the EP study for the ra inbow trout, the endpoint is slightly lower (96-hr LC 50 

### = 4.3 mg a.i./L measured; 95% C.I. 2.9 - 6.3) than the TGAI but the confidence interval overlaps with that of the TGAI. This suggests that the formul ation is not more toxic to rainbow trout. The lowest quantitative 96-hr LC 50 

### for bluegill sunfish was 6.5 mg a.i./L (mean measured, 95% Confidence interval 5.5 -7.7) and for the fathead minnow 8.3 mg a.i./L (measured, 95% C.I. 6.7 – 10.2), both of these were for

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

3 the formulation. Based on test results classifi ed as acceptable, fluridone would be classified as moderately toxic (96-h LC 50 

### >1-10 mg/L) to rainbow trout, channel catfish, bluegill sunfish, and fathead minnow in accordance with EFED’s toxicity classification system.

No replicates were available to determine NOAEC valu es statistically. Sublethal effects of immobility, loss of equilibrium, and surfacing were observed in ra inbow trout exposed to nominal concentrations of 3.2, 3.2, and 5.6 mg a.i./L, respectively. Immobility, loss of equilibrium, and surfacing were also observed in channel catfish, bluegill sunfish, and fa thead minnow but at higher nominal concentrations. Confidence intervals of 96-hr LC 50 

### s overlapped for toxicity tests for the rainbow trout performed at pH of 6.5, 7.5, and 8.5 indicating that pH did not influen ce the toxicity of fluridone. Confidence intervals of qualitative endpoints for the rainbow trout performed at different hardness (40 and 320 mg/L CaCO3) and temperatures (7, 12, and 17 o 

### C). This suggests that hardness and temperature also did not influence the toxicity of fluridone. See 

### Table 17 for results for each test.

In several tests, only binomial 96-hr LC 50 

### values were available because partial mortality was only observed at one treatment level or at no treatment le vels below solubility. The slope of the dose-response curve for fluridone appears to be very steep i ndicating that the distance between the threshold concentration that produces effects and the concentration that will result in 100% mortality is very small. Additionally, the threshold of effects appears to be near the level of solubility for fathead minnow and channel catfish. This results in some difficulty in measuring the median lethal concentration as the exposure concentration must be near solubility to observe partial mortality above and below the LC 50 

### . Many of the tests had measured concentrations at <79% nominal. This was reportly due to storing samples too long and poor handling of the samples. Th e short aqueous photolysis half-life of fluridone indicates that it will be common for fluridone to degrad e over the test duration. Concentrations must be measured in order to verify that the expos ure concentration was constant over time.



### Table 1 summarizes the results of fluridone tests classi fied as acceptable. Table 17 summarizes all the study results and classifications. Rainbow trout test numbers 13, 18, 19, 20, 21, are classified as acceptable and satisfy the data requirement for a T GAI study conducted with a cold water species. Channel catfish test number 32 is classified as accepta ble and satisfies the data requirement for a TGAI study conducted with a warm water species. Observa tion 14, 24, and 36 for the rainbow trout, channel catfish, and bluegill, respectively, are classified as acceptable EP studies for Sonar 4 AS. It is not known if Sonar 4 AS is representative of an EP currently on the market.

Results Synopsis

Test Organism Size (mean wet weight and/or length) and Age : Rainbow Trout fingerlings (0.7 – 1.2 g) Bluegill Sunfish fingerlings (0.5 g) Channel Catfish fingerlings ( 0.4 – 0.9g) Fathead Minnnow fingerlings (0.8 g)

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

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

4 Table 1. Summary of the tests classified as acceptable. Obs

# Test Organism TGAI/ EP Temp., pH, Hardness and Alkalinity as mg/L CaCO 3 96-hr LC 50 , mg a.i./L (95% CI)/ [Slope, (95% CI)] Basis/ Statistical Method Study Classification/ Comments

13 Rainbow Trout, 0.7 g TGAI 12 o C, 7.3 pH, 40, 35 5.1 (4.1-6.4)/

[N/A] Measured/ Binomial Acceptable/ Included only reported measured values to calculate endpoint 14 Rainbow Trout, 1.1 g EP 12 o C, 7.1 pH, 40, 35 4.3 (2.9-6.3)/

[N/A] Measured/ Binomial Acceptable/ Included only reported imeasured values to calculate endpoint 18 Rainbow Trout, 0.98 g TGAI 12 o C, 6.5 pH, 40, 35 5.7 (4.5-7.4)/ [6.7, 3.0-10.3] Nominal*/ Probit Acceptable/ Included only nominal concentrations below the solubility limit. 19 Rainbow Trout, 0.98 g TGAI 12 o C, 7.5 pH, 40, 35 5.6 (3.2-10.0)/

[N/A] Nominal*/ Binomial Acceptable/ Included only nominal concentrations below the solubility limit. 20 Rainbow Trout, 0.98 g TGAI 12 o C, 8.5 pH, 40, 35 7.0 (5.6-10.0)/

[N/A] Nominal*/ Binomial Acceptable/ Included only nominal concentrations below the solubility limit. 21 Rainbow Trout, 0.98 g TGAI 12 o C, 8.1 pH, 40, 235 6.7 (5.3-8.5)/ [7.7, 3.1-12.2] Nominal*/ Probit Acceptable/ Included only nominal concentrations below the solubility limit. 22 Rainbow Trout, 0.98 g TGAI 12 o C, 8.1 pH, 320, 235 8.0 (5.6-10.0)/

[N/A] Nominal*/ Binomial Acceptable/ Included only nominal concentrations below the solubility limit. 24 Fathead Minnow, 0.78 g EP 22 o C, 7.1 pH, 40, 35 8.3 (6.7 – 10.2)/ [N/A] Measured/ Binomial Acceptable/ Study authors endpoint from measured values 32 Channel Catfish, 0.4
g TGAI 22 o C, 8.5 pH, 42, 35 7.5 (6.1-9.2) [10.2, (4.3-16.1)] Nominal*/ Probit Acceptable/ Included only nominal concentrations below the solubility limit. 36 Bluegill, 0.5 g EP 22 o C, 7.1 pH, 40, 35 6.5 (5.5 – 7.7)/ [N/C] Measured/LitchfieldWilcoxon Acceptable/ Measured concentrations were not provided but the 96-hr LC50 based on measured concentrations was reported in Appendix F of the study report. Obs # = M&E (1986) study observation number for fluridone; EP=End-use product; TGAI=Technical Grade Active Ingredient; CI=confidence interval; N/A = not applicable *Nominal exposure levels used in determination of endpoint but nominal con centrations were confirmed analytically (fluridone concentrations average > 79 percent of nominal concentrations).

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

5 I. MATERIALS AND METHODS

GUIDELINE FOLLOWED: The protocol was based on methods recommended by the Committee on Methods for Toxicity Tests with Aquatic Organisms (1975). Methods for acute toxicity tests with fish, macroinvertebrates, and amphibians. U.S. EPA. Ecol. Res. Serv. EPA-660/3-75-009, 61p, and standard protocols of the National Fisheries Research Laboratory

Study Deficiencies As Compared to draft OPPTS 850.1075 Fish Acute Toxicity Guideline : 1. The physiochemical properties of the test material were not reported. However the information was obtained from other sources. 2. A GLP statement was not provided but protocols for recording and documenting test results, study investigators, and similar requirements to GLP were in place. 3. There was only one replicate with 10 individuals at each treatment level, but this did not affect the determination of the EC 50 

### , confidence in estimates may be reduced. 4. Total organic carbon and particulate matter con centrations were not reported but distilled water, which should result in particulate and organic carbon levels to be within acceptable levels, was used for reconstituted water. The well water is assumed to have been within 1975 guideline standards which was cited as being followed. 5. Dissolved oxygen (DO) concentrations were not reported but control mortality was within acceptable levels, indicating DO was not likely an issue. 6. Analytical results for some studies indicate that their nominal concentrations are not appropriate for use as exposure levels. In so me cases analytical results were considered unusable and in others the results were used to calculate study endpoints based on measured levels, these were provided in an appendix whereas the main body of the study reported results based on nominal. In either case the actua l analytical results were not included in the study report. For the specific studies affect ed, these deviations significantly affect the accuracy and reliability of the concentrati on-response relationship and the measurement endpoint value.

Except for the analytical results for some studies, th ese deviations are considered minor for the reasons provided and do not affect the scientific soundness of this study. Specific deficiencies associated with analytical results are discussed below.

Other deficiencies specific to each observation/test. Observation # 15, 16, 17, 25, 26 and 7/17/80 Channel catfish study: These tests were classified as supplemental-qualitative because measured concentrations of soluble fluridone are not available. This is a significant deviation from guidance for low solubility t est materials tested near or above their solubility and for test materials that have short half-lifes, su ch as fluridone’s photodegradation half-life of 0.9-2.3 days, which potentially impact its stability during the test. If fluridone has degraded or become unavailable due to insolubility, the use of nominal con centrations in this case will make fluridone appear less toxic than it really is ( i.e ., LC 50 

### values calculated using nominal con centrations will be higher than if they were based on measured, soluble fluridone con centrations). In a risk assessment use of these values will likely underestimate risk.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

6 Observation # 23, 27, 28, 29, 30, 31, 33, 34, and 35: While the study reported that fluridone concentration average was > 79% of nominal for these studies, it is highly unlikely that the nominal concentrations near and above solubility are representa tive of soluble fluridone concentrations. For these studies knowing the actual measured concentrations for each treatment level is important because effects primarily occured at treatments above known solubility of fluridone (12.3 ppm in distilled water). Some specific treatment measurements, rather than averages, provided in the report for 10, 18, and 56 ppm nominal treatments were 63 to 64% (obs 13, 14) , 37% (obs 24), and 18% (obs 24) of nominal, respectively. Additionally one study reported insol uble fluridone present at nominal 18, 32, and 56 ppm fluridone (obs 25) treatments but identified that fluridone average concentrations > 79% of nominal indicating that specific treatments may have had issues. Therefore, the lack of the actual analytical results for these results introduces a high degree of uncertainty in the representation of nominal concentrations for concentrations near and above solubility even t hough the average concentration for a study is reported
as > 79% of nominal, as individual treatments may be well be low this. Lack of raw data which allows an independent evaluation of the soluble concentrati on at which effects occur under these circumstances is considered a significant deviation. If fluridone h as become unavailable due to insolubility, the use of nominal concentrations in this case will make fl uridone appear less toxic than it really is ( i.e ., LC 50 

### values calculated using nominal concentrations will be highe r than if they were based on measured, soluble fluridone concentrations). In a risk assessment u se of these values will likely underestimate risk. These studies are potentially repairable if the raw measure d fluridone concentrations for each treatment and information about the filter pore size were submitted.

COMPLIANCE : A statement of compliance with GLP procedures was not provided. The studies were completed prior to the GLP requirement. However, protocols for recording and documenting test results, study investigators, measurement of batch test material purity, measurement of test material during the study, and similar requirements to GLP appear to have been in place.

A. MATERIALS :

1. Test Material TGAI: Technical grade Fluridone (purity: 98% active) EP : Sonar 4 AS , 4 lb a.i./gallon field formulation (purity: 48% fluridone)

Description: TGAI: powder EP: liquid

Lot No./Batch No. : TGAI: D36-Y93-107 (Page 389) EP: X-36153 (Page 388)

Purity: TGAI: 98% EP: 48%

Stability of Compound Under Test Conditions: Except for the 7/17/80 channel catfish study, sampling was conducted on all treatment levels at 2 sampling intervals (p 342 of study report), this w ould be consistent with

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

7 sampling at test initiation and termination. Raw flur idone concentration data were not provided but the results were summarized on Page 405 of the study repor t and summarized below in Table 2. As fluridone is expected to degrade rapidly in the presence of li ght (aqueous photolysis half-life = 0.9-2.3 days; MRID 00103326) and tests included nominal concentrations near and above known fluridone solubility, measured concentrations of soluble fluridone c oncentrations are necessary for the TGAI studies to document that exposure to soluble fluridone concentr ations were stable throughout the duration of the study. For studies with the EP measurement of the soluble fluridone concentrations are necessary to document exposure to bioavailable fluridone.

Table 2. Summary of analytical measurements of th e test concentrations and stability conclusions.

Test Species Observation Number Summary of Analytical Results* Are the exposure concentrations reliable?

Observation 13, 14, 24 Analtyical results indicate that actual fluridone concentrations averaged <79% nominal. Measured values were reported for highest treatment with no mortality and lowest treatment with 100% mortality at 96 hours. Nominal values are not reliable but measured values were provided which allow calculation of endpoint using binomial method.

Observation 23, 27, 28, 29, 30, 31 Measured concentrations averaged ≥ 79% of nominal concentrations. While the averaged fluridone concentrations indicate reliability, specific exposure concentrations above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24) and insoluble fluridone reported in obs 25. For these studies effects occurred primarily above solubility. Without actual reported analytical results for these specific treatments, nominal results are considered highly uncertain as representative of soluble exposure concentrations and are qualified either on a total basis or if no mortality occurred at the highest treatment below solubility, it is assigned as a suitable limit test. Observation 15, 16, 17, 25, 26, and Channel catfish study not reported in M&E (1986) Analysis of test material was not performed. Water samples were not collected (channel catfish), lost (obs 25, 26), or samples were stored longer than 6 months and the sample shipment was received in poor condition (obs 15, 16, 17). Nominal concentrations are not considered a reliable quantitative representation of soluble exposure concentrations throughout the study, due to potential for degradation and insolubility.

Observation 18, 19, 20, 21, 22, 32, 33, 34, 35 Measured concentrations indicate that actual fluridone concentrations
average ≥ 79% of nominal. Given that mortality started occurring primarily below known fluridone solubility, and concerns with including results above known solubility without actual raw measured fluridone concentrations, these treatments were dropped by the reviewer from calculation of toxicity endpoints. The

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

8 Test Species Observation Number Summary of Analytical Results* Are the exposure concentrations reliable?

test material concentrations below solubility were considered stable and nominal concentrations representive of exposure within current guideline standards.

Observation 36 Measured concentrations were <79% of nominal. Nominal concentrations are not reliable but the study authors calculated the toxicity endpoint based on measured concentrations, which would be appropriate and within current guideline standards. *From pages 405-408 of study report. 

### ( EPA test guidance recommends that test material concentration not decline to less than 80% of the initial (day-0) measured conc entration and the ratio of the highest concentration to the lowest concentration should be at or less than 1.5:1 among replicates of the same treatment level.) 

### ]

Storage conditions of test material: Not reported.

Table 3. Physicochemical properties of fluridone. Parameter Values Comments

Water solubility at 20oC 12.3 mg/L at 25 o C Footprint @ http://sitem.herts.ac.uk/aeru/footprint/en/ , accessed on April 28, 2009
Vapor pressure 9.77 Η 10 -8
<1 Η 10 -7 USEPA 2007c Weed Science, 4 th Ed. 2002 UV absorption Not reported

pK a 1.73* R. Grover and A. J. Cessna. 1991. Environmental Chemistry of Herbicides Volume II, CRC Press

pK b 12.3* Mackay, D., Shiu, W. Y., Ma, K. C., & Lee, S. C. 2006. Handbook of Physical-Chemical Properties and Environmental Fate for Organic Chemicals (Second ed. Vol. IV). Boca Raton, FL: CRC Press.

K ow 74.13 Log K ow 1.87 from Footprint @ http://sitem.herts.ac.uk/aeru/footprint/en/ , accessed on April 28, 2009 Aquatic photolysis half-life 0.9-2.3 days MRID # 103326 (EPA/OECD guidance recommends reporting water solubility under test conditions, stability in water and light, pKa, Pow, and vap or pressure of test material..) * Therefore, the unionized form should be present within the environmentally relevant pH range.



### 2. Test organism :

Species: Rainbow Trout ( Oncorhynchus mykiss formerly Salmo gairdneri ) Bluegill Sunfish ( Lepomis macrochirus ) Channel Catfish ( Ictalurus punctatus ) Fathead Minnnow ( Pimephales promelas )

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

9 (EPA test guideline preference (850.1075 and 40CFR158) is Rainbow Trout (Oncorhynchus mykiss) for a cold water species, is Blue gill Sunfish (Lepomis macrochirus) for a warm water species, and silverside Me dnida species (Menedia menedia, M. peninsulae, or M. beryllina ) for estuarine/marine species. Other fresh water test guideline speci es include Atlantic Salmon, Salmo salar; Brook Trout, Salvelin us fontinalis; Channel Catfish, Ictalurus punctatus; Coho Sal mon, Oncorhynchus kisutch; Common Carp , Cyprinus carpio; Fathead Minnnow, Pimepha les promelas; Guppy, Poecilia reticulata; Medaka, Oryzias latipes; and Zebra Fish, Brachydanio rerio. Another estuarine/marine tes t guideline species includes the Sheepshead Minnow, Cyprinodon variegates.)


### Age at test initiation : Rainbow Trout (RBT) fingerlings Bluegill Sunfish (BG) fingerlings Channel Catfish (CCF) fingerlings Fathead Minnnow (FHM) fingerlings (EPA/OECD guideline age is juvenile fish and should be the same for all test organisms at test initiation)



### Weight at study initiation: RBT (mean 0.7 – 1.2 g) BG (mean 0.5 g) CCF (mean 0.4 – 0.9g) FHM (mean 0.8 g) (EPA guideline is that juvenile fish <3.0 g should be used)



### Length at study initiation: Not reported (EPA guidance recommends the longest should not be more than twi ce the length of the shortest; OECD guidance recommends length at study initiation of 2.0 ∀ 1.0 cm for Bluegill, Guppy, Fathe ad Minnow, Medaka, and Zebra Fish; 3.0 ∀ 1.0 cm for Common Carp; and 5.0 ∀ 1.0 cm for Rainbow Trout.)



### Source: Laboratory cultures (fathead minnow and channel catfish) or federal fish hatcheries (rainbow trout and bluegill)

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

10 B. STUDY DESIGN :

1. Experimental Conditions

a) Range-finding Study: None reported in study report.

b) Definitive Study



### Table 4. Experimental Parameters
Remarks

Parameter Details Guidance
Acclimation: Period: Conditions: (same as test or not)

Feeding: Health: (any mortality observed) 3-4 days Water was gradually changed from well water to reconstituted test water. Alkalinity was reduced from 237 mg/L CaCO 3 in well water to 35 mg/L as CaCO 3 in the standard, temperature and pH test series and 3.5 mg/L in one CCF test (was 235 mg/L for hardness test series and for RBT temperature test series). Hardness was reduced from 272 mg/L CaCO 3 in well water to 40* mg/L as CaCO 3 in the standard, pH, and temperature test series and one series of hardness tests but was increased to 320 mg/L as CaCO 3 in the other. (*42 mg/L in the CCF pH test series). For RBT, temperature was reduced from 17 o C to 12 o C for standard, pH and hardness test series, and from 17 o C to 7

o C, 17 o C to 12 o C for temperature series. For CCF holding temperature was reduced from 17 o C to 12 o C, and increased from 17 o C to 22 o C for the temperature series. For BG, temperature was increased from 17 o C to 22 o C.

Not fed during acclimation (3-4 days) Not reported EPA/OECD guidance recommends a minimum 12-day holding period with at least 7 days of this period used for acclimation to test conditions. EPA/OECD test guidance is for test fish not to be fed 24 hours prior to test initiation. If fish are larger than 0.5 g EPA guidance recommends that this be extended to 48 hours.

Duration of the test 96-hours EPA/OECD test guidance is a minimum of 96 hours.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

11 Remarks Parameter Details Guidance
Test condition:Static/Static Renewal/Flowthrough

Type of dilution system for flowthrough method

(Flow rate for flow-through system or Renewal rate for static renewal) Static

Not applicable

Not applicable EPA recommends a minimum flow rate of 5 volume additions per 24 hours. To document reproducibility and stability (+ 10%) meter systems should be calibrated before the study and checked twice daily during the test period.

Aeration, if any None

EPA/OECD guidance allows aeration in test chambers in static systems with increased analyses of test material to document itsstability, but it is not recommended in flowthrough systems. Loading in some test vessels exceeded OPPTS guideline recommendations but not OECD guideline recommendations; control mortality levels were acceptable indicating that the loading did not cause adverse effects on fish. Test vessel: Material: Size: Fill volume: wide mouth glass jars 18.9 L (5 gallon) 15 L of test solution

Loading: ranged from 0.3 g/L to 0.8 g/L

Meets recommended loading. EPA recommends loading in static/static renewal of < 0.5 g wet weight/L and in flow-through systems < 0.5 g wet weight/L/24 hours. OECD recommends a maximum of 1 g fish/L for static and semi-static with higher rates accepted for flow-through Source of dilution water Reconstituted water made from deionized water of at least 10 6 ohms resistivity and addition of reagent grade chemicals. Water was mixed thoroughly and aerated before use. EPA prefers soft reconstituted water or water from a natural source not dechlorinated tap water. However, use of dechlorinated tap water may be supportable if the biological responses for the organisms and chemical analyses of residual chlorine meet conditions in the Agency’s 850.1010 guidance for dilution water (http: //www.epa.gov/opptsfrs/ publications/OPPTS_Harmonized/ 850_Ecological_Effects_Test_Guidelines/ Drafts/850-1010.pdf)

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

12 Remarks Parameter Details Guidance Water parameters : Hardness (freshwater)

pH

Dissolved oxygen Total organic carbon

Particulate matter

Metals Pesticides Chlorine Alkalinity

Conductivity Chemical oxygen demand (when testing cationic materials)

Temperature

Intervals of water quality
measurement 40, 42, 43, and 320 mg/L as CaCO 3 in 19, 3, 1, and 3 tests, respectively (see Table 1) See Table 1 for details Standard and temperature test series: 7.1- 7.5 pH test series: 6.5, 7.5, 8.5 (one 8.5 test with CCF varied from 7.9 – 8.65, the test was repeated) Hardness test series: 8.1 Not reported Not reported, but deionized water should be within acceptable levels Not reported, but deionized water should be within acceptable levels

Within acceptable levels <0.01 mg/L Not applicable 3.5, 35, and 235 mg/L as CaCO 3 in 1, 17, and 7 tests, respectively (see Table 1)

< 10 6 ohms resistivity Not applicable

See Table 1 for specific tests RBT 12 o C for all test series, and 7 and 17

o C added for temperature series. BG and FHM 22 o C CCF 22 o C for all test series, and 7 and 17

o C added for temperature series. Monitoring not reported but based on protocols followed temperatures would have been maintained within + 1 o C during the tests.

Standard reconstituted water recipe was used, monitoring of DO, and pH during the test was not reported, except for pH in pH test series which is reported every 24 hours. Hardness, pH and alkalinity would be at least at test initiation. EPA test guidance consists of: hardness : <250 mg/l as CaCO 3 (40 – 48 mg/l CaCO 3 for metals) (OECD guidance is 10 – 250 mg/l CaCO 3 ) pH: 6.0 – 8.5 for freshwater fish with preferably 7.2-7.6; 7.5-8.5 for saltwater fishwith preferably 8.0-8.3 for marinestenohaline fishes; 7.7-8.0 for estuarineeuryhaline fishes (OECD guidance is 6-8.5) Temperature : 22 o C is preferred for the warm water fish: Bluegill, Channel Catfish, Common Carp, Guppy, Medaka, Sheepshead Minnow and the silversides; 12 o C is preferred for the cold water fish: Atlantic Salmon, Brook Trout, Coho Salmon, Rainbow Trout, and Zebra Fish (OECD temperature guidance is 13 – 17 o C for Rainbow Trout, 20 – 24 o C for Common Carp, and 21 – 25 for Bluegill, Fathead Minnow, Guppy, Medaka, and Zebra Fish.) During a given test the selected temperature should be constant within ± 1 o C (OECD recommends constant within + 2 o C) Dissolved oxygen : 60 - 100% saturation (OECD guidance is > 60% saturation) Total organic carbon : guidance is < 2 mg/L (OECD guidance is also < 2 mg/L) Salinity : recommended is 30-34 parts per thousand; weekly range < 6 %)

Treatment concentrations : Nominal Concentrations:

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

13 Remarks Parameter Details Guidance Nominal, corrected for purity:

Measured: See Tables 6, 7, 8, and 9 for specific tests. All tests included 3.2, 5.6, 10, 18, and 32 mg a.i./L nominal (RBT tests also included 1.8 and 56; 1.0; and 100 mg a.i./L in 9, 8, and 1 test(s), respectively, out of the 10 RBT tests) (FHM tests also included 0.56 and 1.0; 56; and 100 mg a.i./L in 1, 2, and 1 test(s), respectively, out of the 2 FHM tests)— same holds for the 2 BG tests (CCF tests also included 0.56; 1.0; 1.8; 56; 100; and 4.2, 7.5, 13.5, and 24 in 6, 7, 8, 2, 1 and 1 test(s), respectively, out of the 11 CCF tests)

TGAI tests included both a solvent (acetone) and a negative control

EP tests did not use an additional solvent for stock solutions or direct additions, which is appropriate, so only negative controls were tested

See Tables 1, 2, and 6 for details EPA/OECD guidance for a definitive test is at least 5 treatment levels in a geometric series which bracket the desired LC 50 plus a dilution control and a solvent control, if a solvent was used. EPA prefers the spacing factor be at least 1.6 and OECD prefers the spacing factor not exceed 2.2.

Solvent (type and concentration, if used) TGAI tests: Acetone 0.5 mL/L for fish

Additional solvent not used for EP tests EPA/OECD recommends that solvents should not exceed 0.1 ml/l Number of replicates: Control (dilution water): Solvent control: Treatments: None EPA/OECD guidance is for no replicates but EPA prefers a minimum of 2.

Number of organisms per replicate : Control (dilution water): Solvent control: Treatments: 10 10 10 EPA/OECD guidance recommends a minimum number of 7 organisms per replicate. EPA prefers a minimum of 10 fish per replicate.

Lighting 16 hours light:8 hours dark

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

14 Remarks Parameter Details Guidance Laboratory lighting using a combination of Duro-test and wide-spectrum Growlux bulbs were reported for chronic testing at the lab and may have been used for acute testing. Lighting parameters were not reported; however, the lighting set up should be consistent with the ASTM 1980 testing parameters and the Committee on Methods for Toxicity Tests with Aquatic Organisms (1975). EPA/OECD recommended photoperiod regime is selected from 12 – 16 hours of light with the selected regime constant during test.

Feeding Not fed EPA/OECD test guidance is for test organisms not to be fed during the study. Recovery of test material: Method recoveries and frequency of determination

% of Nominal Level of Quantitation Level of Detection A coefficient of variation of 1.8 (N=8) and 3.5% (N=8) was obtained for control pond water fortified with 1.0 ppm and 0.05 ppm of fluridone, respectively

See Table 2. The method was reportedly able to detect ~0.005 – 0.001 mg fluridone/L Positive control (if used, indicate the reference test material and concentrations} Not applicable

Other parameters, if any



### 2. Observations :

Table 5. 

### Observations Remarks

Parameter Details Guidance

Parameters measured including the sublethal effects/toxicity symptoms Observed for survival, loss of equilibrium, immobility, surfacing, labored respiraton, rapid respiration, pigmentation changes
Observation intervals 24-hours

EPA/OECD recommends that observation intervals be a minimum of every 24 hours.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

15 Remarks Parameter Details Guidance

Were raw data included? Mortality and sublethal effects raw data was included in the Federal Lab data sheets but analytical test material concentration raw data were not included. Other observations, if any



### II. RESULTS and DISCUSSION:

A. MORTALITY : No mortality/immobility was observed in any of the cont rols. No replicates were available to determine NOAEC values statistically. Mortality was observed in all tests at some treatment level. Many treatment levels were greater than solubility and were not used in statistical analysis because the exposure concentration is unknown. This results in some test s not having enough treatment levels to statistically analyze the results. (EPA/OECD test guidance is < 10% control mortality.)

Table 6. Effect of fluridone on morta lity of freshwater fish Rainbow Trout. (*Treatment levels are above solubility (12.3 mg/L). **Measured fl uridone concentrations. ~Analytical fluridone average concentration > 79% nominal. Treatments in bold were used by reviewer to calculate toxicity endpoint.) Cumulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatment (mg a.i./L) Nominal pH Hardness as mg/L CaCO 3 Temp

o C No. of Fish at

Test Initiation No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dead % Mort Obs 13 7.3 40 12 Observation 13, TGAI, Reconstituted Water 32* 10 7 70 8 80 9 90 10 100 18* 10 3 30 7 70 9 90 10 100 10 ( 6.4 **) 10 2 20 5 50 10 100 10 100 5.6 ( 4.1 **) 10 0 0 0 0 0 0 0 0
3.2 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs 14 7.1 40 12 Observation 14, EP, Reconstituted Water 100* 10 10 100 10 100 10 100 10 100 56* 10 8 80 10 100 10 100 10 100 32* 10 3 30 9 90 10 100 10 100 18* 10 0 0 8 80 10 100 10 100 10 ( 6.3 **) 10 0 0 1 10 8 80 10 100

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

16 Cumulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatment (mg a.i./L) Nominal pH Hardness as mg/L CaCO 3 Temp

o C No. of Fish at

Test Initiation No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dead % Mort
5.6 10 0 0 0 0 0 0 3 30 3.2 ( 2.9 **) 10 0 0 0 0 0 0 0 0
1.8 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs 15 7.5 40 7 Observation 15, TGAI, Reconstituted Water 56* 10 10 100 10 100 10 100 10 100 32* 10 10 100 10 100 10 100 10 100 18* 10 3 30 8 80 9 90 10 100
10 10 1 10 7 70 7 70 9 90
5.6 10 0 0 0 0 0 0 0 0
3.2 10 0 0 0 0 0 0 0 0
1.8 10 0 0 0 0 0 0 0 0
1.0 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs 16 7.5 40 12 Observation 16, TGAI, Reconstituted Water 56* 10 10 100 10 100 10 100 10 100 32* 10 10 100 10 100 10 100 10 100 18* 10 10 100 10 100 10 100 10 100
10 10 5 50 8 80 9 90 10 100
5.6 10 4 40 4 40 5 50 9 90
3.2 10 0 0 0 0 0 0 0 0
1.8 10 0 0 0 0 0 0 0 0
1.0 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs 17 7.5 40 17 Observation 17, TGAI, Reconstituted Water 56* 10 10 100 10 100 10 100 10 100 32* 10 10 100 10 100 10 100 10 100 18* 10 10 100 10 100 10 100 10 100
10 10 10 100 10 100 10 100 10 100
5.6 10 2 20 6 60 8 80 10 100
3.2 10 0 0 0 0 0 0 0 0
1.8 10 0 0 0 0 0 0 0 0
1.0 10 0 0 0 0 0 0 0 0

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

17 Cumulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatment (mg a.i./L) Nominal pH Hardness as mg/L CaCO 3 Temp

o C No. of Fish at

Test Initiation No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dead % Mort Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs 18 6.5 40 12 Observation 18, TGAI, Reconstituted Water 56*~ 10 10 100 10 100 10 100 10 100 32*~ 10 10 100 10 100 10 100 10 100 18*~ 10 2 20 7 70 10 100 10 100
10~ 10 4 40 7 70 9 90 9 90
5.6~ 10 0 0 2 20 4 40 6 60
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs 19 7.5 40 12 Observation 19, TGAI, Reconstituted Water 56*~ 10 10 100 10 100 10 100 10 100 32*~ 10 7 70 8 80 8 80 9 90 18*~ 10 3 30 7 70 9 90 9 90
10~ 10 0 0 9 90 10 100 10 100
5.6~ 10 0 0 3 30 4 40 5 50
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs 20 8.5 40 12 Observation 20, TGAI, Reconstituted Water 56*~ 10 4 40 10 100 10 100 10 100 32*~ 10 8 80 10 100 10 100 10 100 18*~ 10 3 30 9 90 10 100 10 100
10~ 10 0 0 6 60 9 90 10 100
5.6~ 10 0 0 0 0 0 0 1 10
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

18 Cumulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatment (mg a.i./L) Nominal pH Hardness as mg/L CaCO 3 Temp

o C No. of Fish at

Test Initiation No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dead % Mort
Obs 21 8.1 40 12 Observation 21, TGAI, Reconstituted Water 56*~ 10 10 100 10 100 10 100 10 100 32*~ 10 10 100 10 100 10 100 10 100 18*~ 10 0 0 0 0 8 80 9 90
10~ 10 0 0 4 40 6 60 9 90
5.6~ 10 0 0 0 0 2 20 3 30
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs 22 8.1 320 12 Observation 22, TGAI, Reconstituted Water 56*~ 10 10 100 10 100 10 100 10 100 32*~ 10 8 80 9 90 10 100 10 100 18*~ 10 2 20 4 40 8 80 10 100
10~ 10 0 0 1 10 8 80 9 90
5.6~ 10 0 0 0 0 0 0 0 0
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Footnotes: Data were obtained from Fe deral Lab Data submitted to support th e Mayer and Ellersieck (1986) data.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

19 Table 7. Effect of fluridone on mort ality of freshwater fish Fathead Minnow. (*Treatment levels are above solubility (12.3 mg/L). **Measured fl uridone concentrations. ~Analytical fluridone average concentration > 79% nominal. Treatments in bold were used by reviewer to calculate toxicity endpoint.) Cumulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatmen t (mg a.i./L) Nominal pH Hardness , CaCO3 Temp

o C No. of Fish at

Test Initiatio

n No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dead % Mort
Obs. 24 7.1 40 22 Observation 24, EP, Reconstituted Water 100* 10 10 100 10 100 10 100 10 100 56* ( 10.2 **) 10 10 100 10 100 10 100 10 100

32* 10 0 0 1 10 1 10 2 20 18* ( 6.7 **) 10 0 0 0 0 0 0 0 0

10 10 0 0 0 0 0 0 0 0
5.6 10 0 0 0 0 0 0 0 0
3.2 10 0 0 0 0 0 0 0 0
1.8 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 23 7.1 40 22 Observation 23,TGAI, Reconstituted Water
32 *~ 10 10 100 10 100 10 100 10 100
18 *~ 10 0 0 0 0 0 0 1 10
10 ~ 10 0 0 0 0 0 0 0 0
5.6~ 10 0 0 0 0 0 0 0 0
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0
0.56~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs.=observation Footnotes: Data were obtained from Fe deral Lab Data submitted to support th e Mayer and Ellersieck (1986) data.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

20 Table 8. Effect of fluridone on morta lity of freshwater fish Channel Catfish. (*Treatment levels are above solubility (12.3 mg/L). ~Analytical fluridone average concentration > 79% nominal. Treatments in bold were used by reviewer to calculate toxicity endpoint. pt = precipitate observed) Cummulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatment (mg a.i./L) Nominal pH Hardness, CaCO3 Temp

o C No. of Fish at

Test Initiatio

n No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dea

d % Mort

Obs. 25 7.1 40 22 Observation 25, TGAI, Reconstituted Water 56*, pt 10 10 100 10 100 10 100 10 100 32*, pt 10 10 100 10 100 10 100 10 100 18 *, pt 10 10 100 10 100 10 100 10 100 10 10 0 0 0 0 0 0 0 0 5.6 10 0 0 0 0 0 0 0 0
3.2 10 0 0 0 0 0 0 0 0
1.8 10 0 0 0 0 0 0 0 0
1.0 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 26 7.1 40 22 Observation 26, EP, Reconstituted Water 100* 10 10 100 10 100 10 100 10 100 56* 10 10 100 10 100 10 100 10 100 32* 10 4 40 10 100 10 100 10 100 18 * 10 0 0 10 100 10 100 10 100 10 10 0 0 0 0 0 0 0 0 5.6 10 0 0 0 0 0 0 0 0 3.2 10 0 0 0 0 0 0 0 0 1.8 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 27 7.1 40 12 Observation 27, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100 18 *~ 10 9 90 10 100 10 100 10 100 10~ 10 0 0 0 0 2 20 5 50 5.6~ 10 0 0 0 0 0 0 0 0 3.2~ 10 0 0 0 0 0 0 0 0 1.8~ 10 0 0 0 0 0 0 0 0 1.0~ 10 0 0 0 0 0 0 0 0 0.56~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 28 7.1 40 17 Observation 28, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

21 Cummulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatment (mg a.i./L) Nominal pH Hardness, CaCO3 Temp

o C No. of Fish at

Test Initiatio

n No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dea

d % Mort

18 *~ 10 10 100 10 100 10 100 10 100 10 ~ 10 0 0 0 0 1 10 1 10 5.6 ~ 10 0 0 0 0 0 0 0 0 3.2~ 10 0 0 0 0 0 0 0 0 1.8~ 10 0 0 0 0 0 0 0 0 1.0~ 10 0 0 0 0 0 0 0 0 0.56~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 29 7.1 40 22 Observation 29, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100
18 *~ 10 2 20 9 90 10 100 10 100
10 ~ 10 0 0 0 0 0 0 0 0
5.6~ 10 0 0 0 0 0 0 0 0
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0
0.56~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 30 6.5 42 22 Observation 30, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100
18 *~ 10 0 0 4 40 10 100 10 100
10 ~ 10 0 0 0 0 0 0 1 10
5.6 ~ 10 0 0 0 0 0 0 0 0
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0
0.56~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 31 7.5 42 22 Observation 31, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100
18 *~ 10 0 0 10 100 10 100 10 100

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

22 Cummulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatment (mg a.i./L) Nominal pH Hardness, CaCO3 Temp

o C No. of Fish at

Test Initiatio

n No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dea

d % Mort

10 ~ 10 0 0 0 0 0 0 0 0
5.6~ 10 0 0 0 0 0 0 0 0
3.2~ 10 0 0 0 0 0 0 0 0
1.8~ 10 0 0 0 0 0 0 0 0
1.0~ 10 0 0 0 0 0 0 0 0
0.56~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 32 8.5 42 22 Observation 32, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100 18*~ 10 1 10 1 10 5 50 10 100
10 ~ 10 0 0 2 20 6 60 9 90
5.6 ~ 10 0 0 0 0 0 0 1 10
3.2 ~ 10 0 0 0 0 0 0 0 0
1.8 ~ 10 0 0 0 0 0 0 0 0
1.0 ~ 10 0 0 0 0 0 0 0 0
0.56 ~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 33 8.1 40 22 Observation 33, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100
18 *~ 10 7 70 8 80 10 100 10 100
10 ~ 10 0 0 0 0 0 0 6 60
5.6 ~ 10 0 0 0 0 0 0 0 0
3.2~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 34 8.1 320 22 Observation 34, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100
18 *~ 10 6 60 10 100 10 100 10 100
10 ~ 10 0 0 0 0 0 0 0 0
5.6~ 10 0 0 0 0 0 0 0 0
3.2~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

23 Cummulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatment (mg a.i./L) Nominal pH Hardness, CaCO3 Temp

o C No. of Fish at

Test Initiatio

n No. Dead % Mort No. Dead % Mort No. Dead % Mort No. Dea

d % Mort

Control (Dilution Water Only) 10 0 0 0 0 0 0 0 0

7/17/80 8.5 430 22 Observation 7/17/80, TGAI, Reconstituted Water 24* 10 10 100 10 100 10 100 10 100 18* 10 10 100 10 100 10 100 10 100 13.5* 10 3 30 10 100 10 100 10 100
10 10 0 0 2 20 7 70 10 100
7.5 10 0 0 0 0 0 0 0 0
5.6 10 0 0 0 0 0 0 0 0
4.2 10 0 0 0 0 0 0 0 0
3.2 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. = observation Footnotes: Data were obtained from Fe deral Lab Data submitted to support the Mayer and Ellersieck (1986) data.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

24 Table 9. Effect of fluridone on morta lity of freshwater fish Bluegill Sunfish. (*Treatment levels are above solubility (12.3 mg/L). **Measured fl uridone concentrations. ~Analytical fluridone average concentration > 79% nominal. Treatments in bold were used by reviewer to calculate toxicity endpoint.) Cumulative Mortality at Observation Period
24-hr 48-hr 72-hr 96-hr Treatmen t (mg a.i./L) Nominal pH Hardness as CaCO 3 Temp

o C No. of Fish at

Test Initiatio

n No. Dea

d % Mort No. Dead % Mort No. Dead % Mort No. Dead % Mort

Obs. 36 7.1 40 22 Observation 36, EP, Reconstituted Water 100* 10 10 100 10 100 10 100 10 100 56* 10 10 100 10 100 10 100 10 100 32* 10 10 100 10 100 10 100 10 100 18* 10 5 50 9 90 9 90 9 90 10 10 0 0 4 40 4 40 4 40 5.6 10 0 0 0 0 0 0 1 10
3.2 10 0 0 0 0 0 0 0 0
1.8 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs. 35 7.1 40 22 Observation 35, TGAI, Reconstituted Water 32*~ 10 10 100 10 100 10 100 10 100 18 *~ 10 10 100 10 100 10 100 10 100 10 ~ 10 2 20 2 20 2 20 2 20 5.6 ~ 10 1 10 1 10 1 10 1 10 3.2 ~ 10 0 0 0 0 0 0 0 0 1.8 ~ 10 0 0 0 0 1 10 1 10 1.0~ 10 0 0 0 0 0 0 0 0 0.56~ 10 0 0 0 0 0 0 0 0 Solvent Control (acetone) 10 0 0 0 0 0 0 0 0 Negative Control 10 0 0 0 0 0 0 0 0 Obs.=observation Footnotes: Data were obtained from Fe deral Lab Data submitted to support the Mayer and Ellersieck (1986) data.



### B. NON-LETHAL TOXICITY ENDPOINTS:

Immobility, loss of equilibrium, and surfacing were all observed for rainbow trout, channel catfish, bluegill, and fathead minnow. Table 10 summarizes the lowest concentrations where sublethal effects were observed in all tests for each species and Tabl e 11 summarizes the sublethal effects observed in all studies. Rainbow trout were the most sensitive and fa thead minnow and bluegill were the least sensitive. Immobility and loss of equilibrium were observed at 3.2 mg a.i./L and surfacing was observed at 5.6 mg a.i./L. No statistical analysis was performed.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

25 Table 10. Summary of the lowest concentrations th at sublethal effects were observed and the highest concentration in the corresponding test wh ere the sublethal effect was not observed. Statistics could not be completed because no replicates were available.

Study Parameter Immobility Loss of Equilibrium Surfacing Rainbow Trout, TGAI LOAEC 3.2 5.6 5.6 NOAEC 1.8 3.2 3.2 Observation 15 and others 13 17 Rainbow Trout, EP LOAEC 5.6 3.2 0 NOAEC 3.2 1.8 0 Observation 14 14 14 Channel Catfish, TGAI LOAEC 5.6 10 7.5 NOAEC 3.2 5.6 5.6 Observation 27 and others 33 A Bluegill, EP LOAEC 18 18 NA NOAEC 10 10 Observation 36 36 Bluegill, TGAI LOAEC 5.6 NOAEC 3.2 Observation NA NA 35 Fathead Minnow, TGAI LOAEC 18 18 NOAEC 10 10 Observation 23 23 NA Fathead Minnow, EP LOAEC 32 NOAEC 18 Observation 24 NA NA

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

26 Table 11. Sublethal effects (a) of fluridone on freshwater Rainbow trout. (All controls had no sublethal effects. *Treatment levels are above solubility (12.3 mg/L). **Measured fluridone c oncentrations. ~Ana lytical fluridone average concentration > 79% nominal.) Effects at Observation period % Immobilized % Loss of equilibrium (LOE) (# immobilized/# live) (# LOE/# live) Treatment (mg a.i./L)
Nominal 24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr Observation 13, Rainbow Trout, TGAI, pH 7.3, 40 mg/L as Ca CO 3 , 12 o C 32* 100 (3/3) 100 (2/2) 100 (1/1) All dead 0 (0/3) 0 (0/2) 0 (0/1) All dead 18* 100 (7/7) 100 (3/3) 100 (1/1) All dead 0 (0/7) 0 (0/3) 0 (0/1) All dead 10 (6.4**) 100 (8/8) 100 (5/5) All dead All dead 0 (0/8) 0 (0/5) All dead All dead 5.6 (4.1**) 70 (7/10) 80 (8/10) 80 (8/10) 80 (8/10) 10 (1/10) 20 (2/10) 20 (2/10) 0 (0/10)
3.2 0 0 0 0 (0/10) 0 0 0 0 (0/10) Observation 14, Rainbow Trout, EP, pH 7.1, 40 mg/L as CaCO 3 , 12 o C
100* All dead All dead All dead All dead All dead All dead All dead All dead
56* 100 (2/2) All dead All dead All dead 0 (0/2) All dead All dead All dead
32* 100 (7/7) 100 (1/1) All dead All dead 0 (0/7) 0 (0/1) All dead All dead

18* 100 (10/10) 100 (2/2) All dead All dead 0 (0/10) 0 (0/2) All dead All dead

10 (6.3**) 100 (10/10) 100 (9/9) 100 (2/2) All dead 0 (0/10) 0 (0/9) 0 (0/2) All dead

5.6 40 (4/10) ? (b) 70 (7/10) 71 (5/7) 0 0 0 (0/10) 0 (0/7) 3.2 (2.9**) 0 0 0 0 (0/10) 0 0 0 10 (1/10)
1.8 0 0 0 0 (0/10) 0 0 0 0 (0/10) Observation 15, Rainbow Trout, TGAI, pH 7.5, 40 mg/L as CaCO 3 , 7 o C
56* All dead All dead All dead All dead
32* All dead All dead All dead All dead
18* 100 (7/7) 100 (2/2) 100 (1/1) All dead
10 100 (9/9) 100 (3/3) 100 (3/3) 100 (1/1)
5.6 60 (6/10 80 (8/10) 60 (6/10) 60 (6/10)
3.2 0 0 0 10 (1/10)
1.8 0 0 0 0 (0/10)
1.0 0 0 0 0 (0/10) None observed

Observation 16, Rainbow Trout, TGAI, pH 7.5, 40 mg/L as CaCO 3 , 12 o C
56* All dead All dead All dead All dead

32* All dead All dead All dead All dead

18* All dead All dead All dead All dead

10 100 (5/5) 100 (2/2) 100 (1/1) All dead
5.6 100 (6/6) 83 (5/6) 80 (4/5) 100 (1/1) None observed

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

27 Effects at Observation period % Immobilized % Loss of equilibrium (LOE) (# immobilized/# live) (# LOE/# live) Treatment (mg a.i./L)
Nominal 24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr

3.2 10 (1/10) 0 0 0 (0/10)
1.8 0 0 0 0 (0/10)
1.0 0 0 0 0 (0/10) Observation 17, Rainbow Trout, TGAI, pH 7.5, 40 mg/L as CaCO 3 , 17 o C
56* All dead All dead All dead All dead All dead All dead All dead All dead
32* All dead All dead All dead All dead All dead All dead All dead All dead
18* All dead All dead All dead All dead All dead All dead All dead All dead
10 All dead All dead All dead All dead All dead All dead All dead All dead

5.6 88 (7/8) (Surf 1/8) 75 (3/4) 100 (2/2) All dead 0 (0/8) 25 (1/4) 0 (0/2) All dead

3.2 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.8 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.0 0 0 0 0 (0/10) 0 0 0 0 (0/10) Observation 18, Rainbow Trout, TGAI, pH 6.5, 40 mg/L as CaCO 3 , 12 o C
56*~ All dead All dead All dead All dead
32*~ All dead All dead All dead All dead
18*~ 100 (8/8) 100 (3/3) All dead All dead
10~ 100 (6/ 6) 100 (3/3) 100 (1/1) 100 (1/1)
5.6~ 70 (7/10) 88 (7/8) 100 (6/ 6) 100 (4 /4)
3.2~ 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10) None observed

Observation 19, Rainbow Trout, TGAI, pH 7.5, 40 mg/L as CaCO 3 , 12 o C
56*~ All dead All dead All dead All dead
32*~ 100 (3/3) 100 (2/2) 100 (2/2) 100 (1/1)
18*~ 100 (7/7) 100 (3/3) 100 (1/1) 100 (1/1)

10~ 100 (10/10) 100 (1/1) All dead All dead

5.6~ 100 (10/10) 100 (7/7) 100 (6/6) 80 (4/5)

3.2~ 0 10 (1/10) 0 (0/10) 0 (0/10)
1.8~ 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10) None observed

Observation 20, Rainbow Trout, TGAI, pH 8.5, 40 mg/L as CaCO 3 , 12 o C

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

28 Effects at Observation period % Immobilized % Loss of equilibrium (LOE) (# immobilized/# live) (# LOE/# live) Treatment (mg a.i./L)
Nominal 24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr
56*~ 100 (6/6) All dead All dead All dead
32*~ 100 (2/2) All dead All dead All dead
18*~ 100 (7/7) 100 (1/1) All dead All dead

10~ 100 (10/10) 100 (4/4) 100 (1/1) All dead

5.6~ 70 (7/10) 60 (6/10) 70 (7/10) 67 (6/9)
3.2~ 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10) None observed

Observation 21, Rainbow Trout, TGAI, pH 8.1, 40 mg/L as CaCO 3 , 12 o C
56*~ All dead All dead All dead All dead
32*~ All dead All dead All dead All dead

18*~ 100 (10/10) 100 (10/10) 100 (2/2) 100 (1/1)

10~ 100 (10/10) 100 (6/6) 100 (4/4) 100 (1/1)

5.6~ 100 (10/10) 80 (8/10) 88 (7/8) 100 (7/7)

3.2~ 10 (1/10) 0 0 0 (1/10)
1.8~ 0 0 0 0 (1/10)
1.0~ 0 0 0 0 (1/10) None observed

Observation 22, Rainbow Trout, TGAI, pH 8.1, 320 mg/L as CaCO 3 , 12 o C 56*~ All dead All dead All dead All dead
32*~ 100 (2/2) 100 (1/1) All dead All dead
18*~ 100 (8/8) 100 (6/6) 100 (2/2) All dead

10~ 100 (10/10) 100 (9/9) 100 (2/2) 0 (0/1)

5.6~ 80 (8/10) 80 (8/10) 90 (9/10) 90 (9/10)
3.2~ 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (1/10)
1.0~ 0 0 0 0 (1/10) None observed

(a) The only sublethal effects observed consistently were immobili zation and loss of equilibrium. There was one incident of surfacing in one fish, test 17 at the 5.6 mg/L nominal treatment at 24 hours. (b) ? = value on raw data sheet was illegible. Surf = surfacing Footnotes: Data were obtained from Fe deral Lab Data submitted to support the Mayer and Ellersieck (1986) data.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

29 Table 12. Sublethal effects of flurid one on freshwater Fathead Minnow. (All controls had no sublethal effects. *Treatment levels are above solubility (12.3 mg/L). **Measured fluridone c oncentrations. ~Ana lytical fluridone average concentration > 79% nominal.) Effects at Observation period Immobility Loss of equilibrium (% Affected; ) (% Affected) Treatment (mg a.i./L)
Nominal  24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr Observation 23, Fathead Minnow, TGAI, pH 7.1, 40 mg/L as CaCO 3 , 22 o C
32*~ All dead All dead All dead All dead All dead All dead All dead All dead
18*~ 90 (9/10) 90 (9/10) 90 (9/10) 0 (0/9) 0 10 (1/10) 10 (1/10) 0 (0/9)
10*~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
5.6~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
3.2~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
0.56~ 0 0 0 0 (0/10) 0 0 0 0 (0/10) Observation 24, Fathead Minnow, EP, pH 7.1, 40 mg/L as CaCO 3 , 22 o C
100* All dead All dead All dead All dead 56* (10.2**) All dead All dead All dead All dead
32* 90 (9/10) 100 (9/9) 100 (9/9) 0 (0/8) 18* (6.7**) 0 0 0 0 (0/10)
10 0 0 0 0 (0/10)
5.6 0 0 0 0 (0/10)
3.2 0 0 0 0 (0/10)
1.8 0 0 0 0 (0/10) None observed

(a) The only sublethal effects observed were immobilization and loss of equilibrium. Footnotes: Data were obtained from Fe deral Lab Data submitted to support the Mayer and Ellersieck (1986) data.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

30 Table 13. Sublethal effects (a) of fluridone on freshwater Channel Catfish. (*Treatment levels are above solubility (12.3 mg/L). **Measured fl uridone concentrations. ~Analytical fluridone average concentration > 79% nominal.) Effects at Observation period Immobility Surfacing (% Affected; ) (% Affected) Treatment (mg a.i./L)
Nominal  24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr Observation 25, Channel Catfish, TGAI, pH 7.1, 40 mg/L as CaCO 3 , 22 o C 56*, pt All dead All dead All dead All dead 32*, pt All dead All dead All dead All dead 18*, pt All dead All dead All dead All dead
10 80 (8/10) 60 (6/10) 40 (4/10) 0 (0/10)
5.6 0 0 0 0 (0/10)
3.2 0 0 0 0 (0/10)
1.8 0 0 0 0 (0/10)
1.0 0 0 0 0 (0/10) None observed (Loss of equilibrium observed at 10 ppm nominal in 2, 4, and 5 fish out of 10 at 48, 72, and 96 hours, respectively)

Observation 26, Channel Catfish, EP, pH 7.1, 40 mg/L as CaCO 3 , 22 o C
100* All dead All dead All dead All dead
56* All dead All dead All dead All dead

32* 100 6 of 6 All dead All dead All dead

18* 100 10 of 10 All dead All dead All dead

10 0 0 0 0
5.6 0 0 0 0
3.2 0 0 0 0
1.8 0 0 0 0 None observed

Observation 27, Channel Catfish, TGAI, pH 7.1, 40 mg/L as CaCO 3 , 12 o C
32*~ All dead All dead All dead All dead All dead All dead All dead All dead
18*~ 100 (1/1) All dead All dead All dead 0 (0/1) All dead All dead All dead
10~ 90 (9/10) 90 (9/10) 88 (7/8) 80 (4/5) 10 (1/10) 10 (1/10) 13 (1/8) 20 (1/5)

5.6~ 20 (2/10) 20 (2/10) 60 (6/10) 100 (10/10) 0 0 0 0 (0/10)

3.2~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)

0.56~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

31 Effects at Observation period Immobility Surfacing (% Affected; ) (% Affected) Treatment (mg a.i./L)
Nominal  24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr

Observation 28, Channel Catfish, TGAI, pH 7.1, 40 mg/L as CaCO 3 , 17 o C

32*~ All dead All dead All dead All dead All dead All dead All dead All dead
18*~ All dead All dead All dead All dead All dead All dead All dead All dead

10~ 90 (9/10) 100 (10/10) 100 (9/9) 89 (8/9) 10 (1/10) 0 (0/10) 0 (0/9) 11 (1/9)

5.6~ 0 0 100 (10/10) 100 (10/10) 0 0 0 0 (0/10)

3.2~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
0.56~ 0 0 0 0 (0/10) 0 0 0 0 (0/10) Observation 29, Channel Catfish, TGAI, pH 7.1, 40 mg/L as CaCO 3 , 22 o C
32~ All dead All dead All dead All dead All dead All dead All dead All dead
18*~ 100 (8/8) 100 (1/1) All dead All dead 0 (0/8) 0 (0/1) All dead All dead

10~ 70 (7/10) 100 (10/10) 100 (10/10) 100 (10/10) 0 0 0 0 (0/10)

5.6~ 40 (4/10) 80 (8/10) 90 (9/10) 0 (0/10) 0 0 0 80 (8/10)
3.2~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10) 0 0 0 0 (0/10)
0.56~ 0 0 0 0 (0/10) 0 0 0 0 (0/10) Observation 30, Channel Catfish, TGAI, pH 6.5, 42 mg/L as CaCO 3 , 22 o C
32*~ All dead All dead All dead All dead

18*~ 100 (10/10) 100 (6/6) All dead All dead

10~ 0 90 (9/10) 90 (9/10) 0 (0/9)
5.6~ 0 0 0 0 (0/10)
3.2~ 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10)
0.56~ 0 0 0 0 (0/10) None observed

Observation 31, Channel Catfish, TGAI, pH 7.5, 42 mg/L as CaCO 3 , 22 o C
32*~ All dead All dead All dead All dead

18*~ 100 10 of 10 All dead All dead All dead None observed

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

32 Effects at Observation period Immobility Surfacing (% Affected; ) (% Affected) Treatment (mg a.i./L)
Nominal  24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr

10~ 60 6 of 10 60 6 of 10 70 7 of 10 0 (0/10)

5.6~ 0 0 0 0 (0/10)
3.2~ 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10)
0.56~ 0 0 0 0 (0/10) Observation 32, Channel Catfish, TGAI, pH 8.5, 42 mg/L as CaCO 3 , 22 o C
32*~ All dead All dead All dead All dead
18*~ 100 (9/9) 100 (9/9) 100 (5/5) All dead

10~ 100 (10/10) 100 (8/8) 25 (1/4) 0 (0/1)

5.6~ 0 20 (2/10) 30 (3/10) 0 (0/9)
3.2~ 0 0 0 0 (0/10)
1.8~ 0 0 0 0 (0/10)
1.0~ 0 0 0 0 (0/10)
0.56~ 0 0 0 0 (0/10) None observed

Observation 33, Channel Catfish, TGAI, pH 8.1, 40 mg/L as CaCO 3 , 22 o C
32*~ All dead All dead All dead All dead
18*~ 100 (3/3) 100 (2/2) All dead All dead
10~ 90 (9/10) 50 (5/10) 50 (5/10) 0 (0/4)
5.6~ 70 (7/10) 70 (7/10) 70 (7/10) 0 (0/10)
3.2~ 0 0 0 0 (0/10) None observed (LOE observed at 10 ppm nominal in 5 out of 10 fish at 48 and 72 hours)

Observation 34, Channel Catfish, TGAI, pH 8.1, 320 mg/L as CaCO 3 , 22 o C
32*~ All dead All dead All dead All dead
18*~ 100 (4/4) All dead All dead All dead
10~ 80 (8/10) 10 (1/10) 10 (1/10) 0 (0/10)
5.6~ 10 (1/10) 10 (1/10) 10 (1/10) 0 (0/10)
3.2~ 0 0 0 0 (0/10) None observed

7/17/80, Channel Catfish, TGAI, pH 8.5, 43 mg/L as CaCO 3 , 22 o C
24* All dead All dead All dead All dead All dead All dead All dead All dead
18* All dead All dead All dead All dead All dead All dead All dead All dead
13.5* 100 (7/7) All dead All dead All dead 0 (0/7) All dead All dead All dead

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

33 Effects at Observation period Immobility Surfacing (% Affected; ) (% Affected) Treatment (mg a.i./L)
Nominal  24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr

10 30 (3/10) 12 (1/8) 0 (0/3) (LOE 1/3) All dead 0 (0/10) 50 (4/8) 100 (3/3) All dead

7.5 0 0 0 0 (0/10) 0 0 10 (1/10) 0 (0/10)
5.6 0 0 0 0 (0/10) 0 0 0 0 (0/10)
4.2 0 0 0 0 (0/10) 0 0 0 0 (0/10)
3.2 0 0 0 0 (0/10) 0 0 0 0 (0/10) (a) 

### S ublethal effects observed consistented primarily of immobilizati on and surfacing. Loss of equilibrium was observed in three incidents: in observation # 25 at 10 ppm nominal in 2, 4, and 5 fish out of 10 at 48, 72, and 96 hours, respectively; in observ ation # 33 at 10 ppm nominal in 5 out of 10 fish at 48 a nd 72 hours; and in the 7/17/80 test at 10 ppm nominal. LOE = loss of equilibrium Footnotes: Data were obtained from Fe deral Lab Data submitted to support the Mayer and Ellersieck (1986) data.

Table 14. Sublethal effects (a) of fluridone on freshwater Bluegill Sunfish. (*Treatment levels are above solubility (12.3 mg/L). **Measured fl uridone concentrations. ~Analytical fluridone average concentration > 79% nominal.) Effects at Observation period Immobility Loss of equilibrium (% Affected; ) (% Affected) Treatment (mg a.i./L)
Nominal  24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr Observation 36, Bluegill, EP, pH 8.5, 43 mg/L as CaCO 3 , 22 o C
100 All dead All dead All dead All dead All dead All dead All dead All dead
56 All dead All dead All dead All dead All dead All dead All dead All dead
32 All dead All dead All dead All dead All dead All dead All dead All dead
18 0 (0/5) 0 (0/1) 0 (0/1) 0 (0/1) 25 (1/5) 0 (0/1) 0 (0/1) 0 (0/1)
10 10 (1/10) 0 (0/6) 0 (0/6) 0 (0/6) 0 (0/5) 0 (0/6) 0 (0/6) 0 (0/6)
5.6 0 0 0 (0/10) 0 (0/9) 0 0 0 (0/10) 0 (0/9)
3.2 0 0 0 0 (0/10) 0 0 0 0 (0/10)
1.0 0 0 0 0 (0/10) 0 0 0 0 (0/10) Observation 35, Bluegill, TGAI, pH 7.1, 40 mg/L as CaCO 3 , 22 o C
32 All dead All dead All dead All dead
18 All dead All dead All dead All dead
10 0 (0/8) 0 (0/8) 100 (8/8) 0 (0/8)
5.6 0 (0/9) 0 (0/9) 100 (9/9) 0 (0/9)
3.2 0 0 0 0 (0/10)
1.8 None observed

0 0 (0/10) 0 (0/9) 0 (0/9)

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

34 Effects at Observation period Immobility Loss of equilibrium (% Affected; ) (% Affected) Treatment (mg a.i./L)
Nominal  24-hr 48-hr 72-hr 96-hr 24-hr 48-hr 72-hr 96-hr

1.0 0 0 0 0 (0/10)
0.56 0 0 0 0 (0/10) (a) The only sublethal effects observed were immobilization and loss of equilibrium. Footnotes: Data were obtained from Fe deral Lab Data submitted to support the Mayer and Ellersieck (1986) data.



### C. REPORTED STATISTICS : The method used to determine the LC 50 

### values, confidence limits, and slopes were the Litchfield and Wilcoxon (1949) method.

Table 15. Statistics Reported in Study Report. Obs Organism Compound Temp

o C pH Hardness, mg/L as CaCO 3 96-hr LC50 (mg a.i./L) with 95% Confidence

interval Slope LC 50 Method

Standard Test 13 Rainbow Trout TGAI 12 7.1-7.3 40 7.6 (6.0-9.7) 1.31 Litchfield* 14 Rainbow Trout EP 12 7.1-7.3 40 7.1 (5.6-9.1) 1.48 Litchfield* Varied Temperature 15 Rainbow Trout TGAI 7 7.1-7.5 40 7.7 (5.8-10.2) 1.56 Litchfield* 16 Rainbow Trout TGAI 12 7.1-7.5 40 4.5 (3.4-5.9) 1.55 Litchfield* 17 Rainbow Trout TGAI 17 7.1-7.5 40 4.2 (3.4-5.4) 1.31 Litchfield* Varied pH 18 Rainbow Trout TGAI 12 6.5 40-42 5.7 (4.1-7.9) 1.45 Litchfield* 19 Rainbow Trout TGAI 12 7.5 40-42 5.6 (4.8-6.6) 1.20 Litchfield* 20 Rainbow Trout TGAI 12 8.5 40-42 8.4 (6.4-11.1) 1.37 Litchfield* Varied Hardness 21 Rainbow Trout TGAI 12 8.1 40 7.6 (5.7-10.1) 1.58 Litchfield* 22 Rainbow Trout TGAI 12 8.1 320 7.6 (5.9-9.7) 1.32 Litchfield* Standard Tests 23 Fathead Minnow TGAI 22 7.1-7.3 40 22 (17-28) 1.34 Litchfield* 24 Fathead Minnow EP 22 7.1-7.3 40 41(32-52) 1.32 Litchfield* Standard Test 25 Channel Catfish TGAI 22 7.1-7.3 40 13.2 (10.3-17.0) 1.33 Litchfield* 26 Channel Catfish EP 22 7.1-7.3 40 13.2 (10.3-17.0) 1.33 Litchfield* Varied Temperature 27 Channel Catfish TGAI 7 7.1 40 10.0 (7.3-13.6) 1.44 Litchfield* 28 Channel Catfish TGAI 17 7.1 40 10.0 (7.3-13.8) 1.44 Litchfield* 29 Channel Catfish TGAI 22 7.1 40 13.0 (10.2-16.6) 1.32 Litchfield* Varied pH 30 Channel Catfish TGAI 22 6.5 40-42 15.0 (11.3-19.9) 1.38 Litchfield* 31 Channel Catfish TGAI 22 7.5 40-42 13.5 (10.4-17.5) 1.34 Litchfield*

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

35 32 Channel Catfish TGAI 22 8.5 40-42 6.2 (6.2-10.8) 1.56 Litchfield* Varied Hardness 33 Channel Catfish TGAI 22 8.1 40 10.0 (7.3-13.8) 1.44 Litchfield* 34 Channel Catfish TGAI 22 8.1 320 14.0 (11.7-16.8) 1.34 Litchfield* Standard Tests 35 Bluegill TGAI 22 7.1-7.3 40 13.0 (9.9-17.1) 1.56 Litchfield*

36 Bluegill EP 22 7.1-7.3 40 12.0 (8.2-17.5) 1.83 Litchfield*

Obs # = observation number in fluridone of M&E (1986). *Statistics calculated according to Litchfield and Wilcoxon (1949).



### D. VERIFICATION OF STATISTICAL RESULTS BY THE REVIEWER : TOXANAL (2009 version) was used to calculate LC 50 

### values; uses binomial, moving average, and probit methods. The probit method results were used when the probit model fit. In most cases the binomial method which does not require partial kills was used when there we re less than two concentrations with percent dead between 0 and 100%. The output files from the analysis are attached at the end of this evaluation report.

Table 16. Reviewer Calculated Stat istical Endpoints. NOAEC values co uld not be determined because there were no replicates in the study. Observation Period Study Result 24 hrs 48 hrs 72 hrs 96 hrs Observation 13, Rainbow Trout, TGAI, pH 7.3, Hardness 40 mg/L as CaCO 3 , 12 o C

LC 50 (95% CI) in mg a.i./L > 6.4 6.4 5.1 (4.1 – 6.4) 5.1 (4.1 – 6.4) Probit Slope (95% CI) NA NA NA NA

LC 50 Method 2 dead/10 (1) Point Estimate 5 dead/10 (1) Binomial (1) Binomial (1)

Observation 14, Rainbow Trout, EP, pH 7.1, Hardness 40 mg/L as CaCO 3 , 12 o C

LC 50 (95% CI) in mg a.i./L >6.3 > 6.3 4.9 (2.9 - +infinity) 4.3 (2.9 – 6.3) Probit Slope (95% CI) NA NA NA NA LC 50 Method no mortality (1) 1 dead/10 (1) Binomial (1) Binomial (1)
Observation 15, Rainbow Trout, TGAI, pH 7.5, Hardness 40 mg/L as CaCO 3 , 7 o C

LC 50 (95% CI) in mg a.i./L > 10 (total) 8.8 (total) (5.6-infinity (total)) 8.8 (total) (5.6-infinity (total)) 8.0 (total) (5.6-10.0 (total))
Probit Slope (95% CI) NA NA N/A N/A LC 50 Method 1 dead/10 (2) Binomial (2) Binomial (2) Binomial (2) Observation 16, Rainbow Trout, TGAI, pH 7.5, Hardness 40 mg/L as CaCO 3 , 12 o C

LC 50 (95% CI) in mg a.i./L 10 (total) 6.8 (total) (5.2-9.2 (total)) 6.0 (total) (4.7 - 7.8 (total)) 4.5 (total) (3.2 - 5.6 (total))
Probit Slope (95% CI) NA 5.8 (2.5 - 9.2) 6.8 (3.0 - 10.6) NA

LC 50 Method Point Estimate 5 dead/10 (2) Probit (2) Probit (2) Binomial (2)

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

36 Observation Period Study Result 24 hrs 48 hrs 72 hrs 96 hrs Observation 17, Rainbow Trout, TGAI, pH 7.5, Hardness 40 mg/L as CaCO 3 , 17 o C

LC 50 (95% CI) in mg a.i./L 6.7 (total) (3.2 – 10 (total)) 5.2 (total) (3.2-10 (total)) 4.7 (total) (3.2-10 (total)) 4.2 (total) (3.2 – 5.6 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method Binomial (2) Binomial (2) Binomial (2) Binomial (2) Observation 18, Rainbow Trout, TGAI, pH 6.5, Hardness 40 mg/L as CaCO 3 , 12 o C

LC 50 (95% CI) in mg a.i./L > 10 8.0 (6.2-12) 6.3 (5.1-8.1) 5.7 (4.5-7.4)
Probit Slope (95% CI) NA 6.0 (2.0 - 9.9) 7.1 (3.1 - 11.2) 6.7 (3.0 - 10.3) LC 50 Method 1 dead/10 (3) Probit (3) Probit (3) Probit (3) Observation 19, Rainbow Trouth, TGAI,

LC 50 (95% CI) in mg a.i./L >10 6.7 (5.3 - 8.5) 6.0 (3.2 - 10) 5.6 (3.2 - 10)
Probit Slope (95% CI) NA 7.7 (3.1 - 12.2) NA NA

LC 50 Method no mortality (3) Probit (3) Binomial (3) Binomial (3) Observation 20, Rainbow Trout, TGAI, pH 8.5, Hardness 40 mg/L as CaCO 3 , 12 o C

LC 50 (95% CI) in mg a.i./L >10 9.3 (5.6 - +infinity) 8.0 (5.6 - 10) 7.0 (5.6 - 10) Probit Slope (95% CI) NA NA NA NA LC 50 Method no mortality (3) Binomial (3) Binomial (3) Binomial (3) Observation 21, Rainbow Trout, TGAI, pH 8.1, Hardness 40 mg/L as CaCO 3 , 12 o C

LC 50 (95% CI) in mg a.i./L >10 > 10 8.7 (6.7 - 16) 6.7 (5.3 - 8.5)
Probit Slope (95% CI) NA NA 5.1 (1.5 - 8.7) 7.7 (3.1 - 12.2) LC 50 Method no mortality (3) 1 dead/10 (3) Probit (3) Probit (3) Observation 22, Rainbow Trout, TGAI, pH 8.1, Hardness 320 mg/L as CaCO 3 , 12 o C

LC 50 (95% CI) in mg a.i./L >10 > 10 8.3 (5.6 - +infinity) 8.0 (5.6-10) Probit Slope (95% CI) NA NA NA NA LC 50 Method no mortality (3) 1 dead/10 (3) Binomial (3) Binomial (3) Observation 23, Fathead Minnow, TGAI, pH 7.0, Hardness 40 mg/L as CaCO 3 , 22 o C LC 50 (95% CI) in mg a.i./L >18 (total) >18 (total) >18 (total) >10 (total) Probit Slope (95% CI) NA NA NA NA LC 50 Method no mortality (4) no mortality (4) no mortality (4) no mortality (4) Observation 24, Fathead Minnow, TEP, pH 7.1, Hardness 40 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L 8.3 (6.7 – 10.2) 8.3 (6.7 – 10.2) 8.3 (6.7 – 10.2) 8.3 (6.7 – 10.2) Probit Slope (95% CI) NA NA NA NA LC 50 Method Binomial (1) Binomial (1) Binomial (1) Binomial (1)

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

37 Observation Period Study Result 24 hrs 48 hrs 72 hrs 96 hrs Observation 25, Channel Catfish, TGAI, pH 7.1, Hardness 40 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method Binomial (4) Binomial (4) Binomial (4) Binomial (4) Observation 26, Channel Catfish, EP, pH 7.1, Hardness 40 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L 14.3 (total) (10 – 18 (total) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method Binomial (4) Binomial (4) Binomial (4) Binomial (4) Observation 27, Channel Catfish, TGAI, pH 7.1, Hardness 40 mg/L as CaCO 3 , 12 o C

LC 50 (95% CI) in mg a.i./L 14.3 (total) (10 -18 (total)) 13 (total) (10 – 18 (total)) 12 (total) (5.6 – 18 (total)) 10
Probit Slope (95% CI) NA NA NA NA

LC 50 Method Binomial (5) Binomial (5) Binomial (5) Point Estimate, 5 dead/10 (3) Observation 28, Channel Catfish, TGAI, pH 7.1, Hardness 40 mg/L as CaCO 3 , 17 o C

LC 50 (95% CI) in mg a.i./L 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method Binomial (5) Binomial (5) Binomial (5) Binomial (5) Observation 29, Channel Catfish, TGAI, pH 7.1, Hardness 40 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L >10 14 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method no mortality (2) Binomial (5) Binomial (5) Binomial (5) Observation 30, Channel Catfish, TGAI, pH 6.5, Hardness 42 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L >10 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method no mortallity (2) Binomial (5) Binomial (5) Binomial (5) Observation 31, Channel Catfish, TGAI, pH 7.5, Hardness 42 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L >10 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method no mortallity (2) Binomial (5) Binomial (5) Binomial (5) Observation 32, Channel Catfish, TGAI, pH 8.5, Hardness 42 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L >10 >10 9.3 (5.6 - +infinity) 7.5 (6.1-9.2)
Probit Slope (95% CI) NA NA NA 10.2 (4.3-16.1) LC 50 Method no mortallity (3) no mortallity (3) Binomial (3) Probit (3)

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

38 Observation Period Study Result 24 hrs 48 hrs 72 hrs 96 hrs Observation 33, Channel Catfish, TGAI, pH 8.1, Hardness 40 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L 16 (total) (10 – 32 (total)) 15 (total) (10 – 32 (total)) 13 (total) (10 – 18 (total)) 9.3 (total) (5.6 – 18 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method Binomial (3) Binomial (3) Binomial (3) Binomial (3) Observation 34, Channel Catfish, TGAI, pH 8.1, Hardness 320 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L 17 (total) (10 - +infinity) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) 13 (total) (10 – 18 (total)) Probit Slope (95% CI) NA NA NA NA LC 50 Method Binomial (5) Binomial (5) Binomial (5) Binomial (5) Observation 35, Bluegill Sunfish, TGAI. pH 7.1, Hardness 40 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L 11 (total) (8.5 – 14 (total)) 11 (total) (8.5 – 14 (total)) 11 (total) (9.0 – 14 (total)) 11 (total) (9.0 – 14 (total)
Probit Slope (95% CI) 6.5 (2.9 – 10.1) 6.5 (2.9 – 10.1) NA NA

LC 50 Method Probit (5) Probit (5) Moving Average (5) Moving Average (5) Observation 36, Bluegill Sunfish, EP, pH 7.1, Hardness 40 mg/L as CaCO 3 , o C

LC 50 (95% CI) in mg a.i./L 6.5 (5.5 – 7.7) 6.5 (5.5 – 7.7) Probit Slope (95% CI) NA NA

LC 50 Method Litchfield- Wilcoxon (6) Litchfield-Wilcoxon (6) 7/17/80, Channel Catfish, TGAI, pH 7.5, Hardness 43 mg/L as CaCO 3 , 22 o C

LC 50 (95% CI) in mg a.i./L 11 (total) (7.5 – 13 (total)) 9.4 (total) (7.5 – 13.5 (total)) 8.7 (total) (7.5 – 10 (total)) LC 50 Method Binomial (4) Binomial (4) Binomial (2) Abbreviations: NA=not applicable (1) Two measured fluridone concentrations were provide d in Appendix F of the USFWS (1981) study report, and only these treatment levels were used to estimate the LC 50 values. Fluridone concentrations averaged <79%, indicating nominal concentrations are not appropriate for use in characterizing the toxicity of fluridone to fish.
(2) Measured concentrations are not availa ble, or the available analytical results indicated that concentrations were not constant over time. Nominal values below known so lubility (12.3 mg/L in distilled water) were used to calculate LC 50 values, but must assume that these represent total concentrations as the solubility under study conditions is unknown, nor if degradation occured.
(3) Fluridone concentrations averaged > 79% of nominal. Nominal values below known solubility (12.3 mg/L in distilled water) were used to calculate LC 50 values, with the analytical agr eement these values are considered representative of soluble fluridone exposure concentrations.
(4) Measured concentrations are not availa ble, or the available analytical results indicated that concentrations were not constant over time. In addition to using nominal values below known sol ubility (12.3 mg/L in distilled water), the treatment just above solubility was also included because the mortality cha nge at this dividing line was great. Results represent total concentrations rather than soluble as the solubility under study c onditions is unknown, nor if degradation occurred.
(5) Mortality occurred at the concentr ation just above known solubility (12. 3 mg/L in distilled water). While fluridone concentrations averaged > 79% of nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nom inal (as evidenced by values that were reported for obs 13, 14, 24) and

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

39 precipitate observed in observation 25. Value above solubility was included in estimate and because of the lack of raw analytical data is therefore reported as total.
(6) Fluridone concentrations averaged <79%. Authors recalculated the 96-h LC50 using measured concentrations rather than nominal and provided the result in Appendix F of the USFWS (1981) report.



### E. STUDY DEFICIENCIES : Study deficiencies identified by this reviewer as compared to current public draft OPPTS 850.1075 fish acute guidelines are discussed in Section I Materials and Methods under Guideline study deficiencies.

F. REVIEWER’S COMMENTS :

G. CONCLUSIONS : Some endpoints were scientifically sound and classified as acceptable, others had deficiencies and were supplemental. Confidence intervals of 96-hr LC 50 

### values overlapped for toxicity tests for the rainbow trout performed at pH of 6.5, 7.5, and 8.5 indicating that pH did not influence the toxicity of fluridone. Confidence intervals of qua litative endpoints for the rainbow trout performed at different hardness (40 and 320 mg/L Ca CO3) and temperatures (7, 12, and 17 o 

### C) overlapped. This suggests that hardness and temperature also did not influence the toxicity of fluridone. Confidence intervals of the TGAI test and the EP test on the rainbow trout overlapped; however, the measured concentrations in the EP study were <79% nominal and it cannot be determined whether the EP was more toxic than the TGAI. See 

### Table 17 for a complete summary of the reviewer determined study results. In several tests, only binomial 96-hr LC 50 

### values were available because partial mortality was only observed at one treatment level or at no treatment levels. The slope of the dose-response curve for fluridone appears to be very steep indicating that the dist ance between the threshold concentration that produces effects and the concentration that will result in 100% mo rtality is very small. Additionally, the threshold of effects appears to be near the level of solubility for fathead minnow and channel catfish. This results in some difficulty in measuring the median lethal concentration as the exposure concentration must be near solubility to observe partial mortality above and below the LC 50 

### . Many of the tests had measured concentrations at <79% nominal. This was reportly due to storing samples too long and poor handling of the samples. The short aqueous photolysis half-lif e of fluridone indicates that it will be common for fluridone to degrade over the test duration. Concentr ations must be measured (in TGAI studies) in order to verify that the exposure concentration was constant over time.

Table 17. Summary of reviewer calculat ed results and study classifications Obs

# Test Organism TGAI/ EP Temp., pH, Hardness and Alkalinity as mg/L CaCO 3 96-hr LC 50 , mg a.i./L (95% CI)/ [Slope, (95% CI)] Basis, Statistical Method Study Classification/ Comments

13 Rainbow Trout, 0.7 g TGAI 12 o C, 7.3 pH, 40, 35 5.1 (4.1-6.4)/

[N/A]/ Measured, Binomial Acceptable/

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

40 Obs

# Test Organism TGAI/ EP Temp., pH, Hardness and Alkalinity as mg/L CaCO 3 96-hr LC 50 , mg a.i./L (95% CI)/ [Slope, (95% CI)] Basis, Statistical Method Study Classification/ Comments

14 Rainbow Trout, 1.1 g EP 12 o C, 7.1 pH, 40, 35 4.3 (2.9-6.3)/

[N/A] Measured, Binomial Acceptable/

Varied Temperature 15 Rainbow Trout, 1.2 g TGAI 7 o C, 7.5 pH, 40, 235 8.0 (total) (5.6-10.0 (total))/ [N/A] Nominal, Binomial Supplemental-qualitative/ Analysis of test material not performed, sample shipment in poor condition and sample storage too long. 16 Rainbow Trout, 1.2 g TGAI 12 o C, 7.5 pH, 40, 235 4.5 (total) (3.2-5.6 (total))/ [N/A] Nominal, Binomial Supplemental-qualitative/ Analysis of test material not performed, sample shipment in poor condition and sample storage too long. 17 Rainbow Trout, 1.2 g TGAI 17 o C, 7.5 pH, 40, 235 7.5 (total) (5.6-10.0 (total))/ [N/A] Nominal, Binomial Supplemental-qualitative/ Analysis of test material not performed, sample shipment in poor condition and sample storage too long. Varied pH 18 Rainbow Trout, 0.98 g TGAI 12 o C, 6.5 pH, 40, 35 5.7 (4.5-7.4)/ [6.7, 3.0-10.3] Nominal*, Probit Acceptable/

19 Rainbow Trout, 0.98 g TGAI 12 o C, 7.5 pH, 40, 35 5.6 (3.2-10.0)/ [N/A] Nominal*,Binomial Acceptable/

20 Rainbow Trout, 0.98 g TGAI 12 o C, 8.5 pH, 40, 35 7.0 (5.6-10.0)/ [N/A] Nominal*, Binomial Acceptable/

Varied Hardness 21 Rainbow Trout, 0.98 g TGAI 12 o C, 8.1 pH, 40, 235 6.7 (5.3-8.5)/ [7.7, 3.1-12.2] Nominal*, Probit Acceptable/

22 Rainbow Trout, 0.98 g TGAI 12 o C, 8.1 pH, 320, 235 8.0 (5.6-10.0)/ [N/A] Nominal*, Binomial Acceptable/

Standard Tests

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

41 Obs

# Test Organism TGAI/ EP Temp., pH, Hardness and Alkalinity as mg/L CaCO 3 96-hr LC 50 , mg a.i./L (95% CI)/ [Slope, (95% CI)] Basis, Statistical Method Study Classification/ Comments

23 Fathead Minnow, 0.78 g TGAI 22 o C, 7.1 pH, 40, 35 >10 (total)

Nominal*, no mortality Supplemental-qualitative/ No mortality at any treatment level below solubility, all mortality occurred above solubility. While fluridone concentration average was reported as > 79% nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Without actual reported analytical results, the results are qualified here as a total basis 24 Fathead Minnow, 0.78 g EP 22 o C, 7.1 pH, 40, 35 8.3 (6.7 – 10.2)/ [N/A] Measured, Binomial Acceptable/

Standard Test 25 Channel Catfish, 0.7
g TGAI 22 o C, 7.1 pH, 40, 35 13 (total) (10 -18 (total)) [NA] Nominal, Binomal Supplemental-qualitative/ Water samples were lost thus no analytical confirmation of soluble exposure concentrations can be made. No mortality occurred at any nominal treatment level below solubility. Exposure concentrations at near solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). 26 Channel Catfish, 0.7
g EP 22 o C, 7.1 pH, 40, 35 13 (total) (10 -18 (total)) [NA] Nominal, Binomal Supplemental-qualitative/ Water samples were lost thus no analytical confirmation of soluble exposure concentrations can be made. No mortality occurred at any nominal treatment level below solubility. Exposure concentrations at near solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Varied Temperature 27 Channel Catfish, 0.9
g TGAI 7 o C, 7.1 pH, 40, 35 10

[NA] Nominal*, Point Estimate 5 dead/10 Supplemental/ Temperature lower than guideline recommendation for this species

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

42 Obs

# Test Organism TGAI/ EP Temp., pH, Hardness and Alkalinity as mg/L CaCO 3 96-hr LC 50 , mg a.i./L (95% CI)/ [Slope, (95% CI)] Basis, Statistical Method Study Classification/ Comments

28 Channel Catfish, 0.9
g TGAI 17 o C, 7.1 pH, 40, 35 13 (total) (10 -18 (total)) [NA] Nominal, Binomal Supplemental-qualitative/ There was one mortality at a treatment level below solubility, all other mortalities occurred in exposure concentrations above solubility. While fluridone concentration average was reported as > 79% nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Without actual reported analytical results, the results are qualified here as a total basis. 29 Channel Catfish, 0.9
g TGAI 22 o C, 7.1 pH, 40, 35 13 (total) (10 -18 (total)) [NA] Nominal, Binomal Supplemental-qualitative/ There was one mortality at a treatment level below solubility, all other mortalities occurred in exposure concentrations above solubility. While fluridone concentration average was reported as > 79% nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Without actual reported analytical results, the results are qualified here as a total basis. Varied pH 30 Channel Catfish, 0.4
g TGAI 22 o C, 6.5 pH, 42, 35 13 (total) (10 – 18 (total))/ [N/A] Nominal, Binomial Supplemental-qualitative/ There was one mortality at a treatment level below solubility, all other mortalities occurred in exposure concentrations above solubility. While fluridone concentration average was reported as > 79% nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Without actual reported analytical results, the results are qualified here as a total basis.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

43 Obs

# Test Organism TGAI/ EP Temp., pH, Hardness and Alkalinity as mg/L CaCO 3 96-hr LC 50 , mg a.i./L (95% CI)/ [Slope, (95% CI)] Basis, Statistical Method Study Classification/ Comments

31 Channel Catfish, 0.4
g TGAI 22 o C, 7.5 pH, 42, 35 13 (total) (10 – 18 (total))/ [N/A] Nominal, Binomial Supplemental-qualitative/ No mortality at any treatment level below solubility, all mortality occurred above solubility. While fluridone concentration average was reported as > 79% nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Without actual reported analytical results, the results are qualified here as a total basis 32 Channel Catfish, 0.4
g TGAI 22 o C, 8.5 pH, 42, 35 7.5 (6.1-9.2) [10.2, (4.3-16.1)] Nominal*, Probit Acceptable/ Included only nominal concentrations below the solubility limit.

7/17/ 80 Channel Catfish, 1.0
g TGAI 22 o C, 7.9 – 8.65 pH, 43, 3.5 8.7 (total) (7.5-10.0 (total))/ [N/A] Nominal, Binomial Supplemental-qualitative/ N o reported analytical sampling indicated for this study. Potential for soluble exposure levels to be lower than nominal. Varied Hardness 33 Channel Catfish, 0.8
g TGAI 22 o C, 8.1 pH, 40, 235 9.3 (total) (5.6-18 (total))/ [N/A] Nominal*, Binomial Supplemental-qualitative/ Mortality occurred near and above solubility. While fluridone concentration average was reported as > 79% nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Without actual reported analytical results, the results are qualified here as on a total basis. 34 Channel Catfish, 0.8
g TGAI 22 o C, 8.1 pH, 320, 235 13 (total) (10 – 18 (total))/ [N/A] Nominal, Binomial Supplemental-qualitative/ No mortality at any treatment level below solubility, all mortality occurred above solubility. While fluridone concentration average was reported as > 79% nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Without actual reported analytical results, the results are qualified here as a total basis

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

44 Obs

# Test Organism TGAI/ EP Temp., pH, Hardness and Alkalinity as mg/L CaCO 3 96-hr LC 50 , mg a.i./L (95% CI)/ [Slope, (95% CI)] Basis, Statistical Method Study Classification/ Comments

Standard Tests 35 Bluegill, 0.5 g TGAI 22 o C, 7.1 pH, 40, 35 11 (total) (9.0 – 14 (total)) [N/A] Nominal, Moving Average Supplemental-qualitative/ Mortality occurred at and above solubility. While fluridone concentration average was reported as > 79% nominal, exposure concentrations at and above solubility are unlikely to have soluble concentrations near nominal (as evidenced by values that were reported for obs 13, 14, 24). Without actual reported analytical results, the results are qualified here as a total basis. 36 Bluegill, 0.5 g EP 22 o C, 7.1 pH, 40, 35 6.5 (5.5 – 7.7)/ [N/C]Measured, LitchfieldWilcoxon Acceptable/ Measured concentrations were not provided but the 96-hr LC50 based on measured concentrations was reported in Appendix F of the study report. Obs # = M&E, 1986 study observation number for fluridone; EP =End-use product; TGAI=Technical Grade Active Ingredient; CI=confidence interval; NR=study not reported in M&E, 1986 *Nominal exposure levels used in determination of endpoint but nominal con centrations were confirmed analytically (fluridone concentrations average > 79 percent of nominal concentrations).



### III. REFERENCES :

Committee on Methods for Toxicity Tests with Aquatic Organisms (1975). Methods for acute toxicity tests with fish, macroinvertebrates, and amphibians. U.S. EPA. Ecol. Res. Serv. EPA-660/3-75- 009. 61p. Litchfield Jr., J. T., & Wilcoxon, F. 1949. A simplif ied method of evaluating dose-effect experiments. J. Pharmacol. Therap., 96 , 99-113.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

45 M and E (1986) Fluridone Obs 13 TGAI 98 percent purity RBT 72 and 96-hr LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 6.4 10 10 100 9.765625E-02 4.1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 4.1 AND 6.4 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 5.122499

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 14 TGAI 98 percent purity RBT 72-hr LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 6.3 10 8 80 5.46875 2.9 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 2.9 AND +INFINITY CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 4.935012

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. ************************************************************

M and E (1986) Fluridone Obs 14 TGAI 98 percent purity RBT 96-hr LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 6.3 10 10 100 9.765625E-02 2.9 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 2.9 AND 6.3 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 4.274342

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

46 M and E (1986) Fluridone Obs 15 TGAI 98 percent purity RBT 48-h and 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 7 70 17.1875 5.6 10 0 0 9.765625E-02 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND +INFINITY CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 8.760268

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICA STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 15 TGAI 98 percent purity RBT 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 10 10 9 90 1.074219 5.6 10 0 0 9.765625E-02 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 7.955181

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

Fluridone Obs 16 TGAI 98 percent purity RBT 48-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 8 80 5.46875 5.6 10 4 40 37.69531 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

47 THE BINOMIAL TEST SHOWS THAT 3.2 AND +INFINITY CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 6.436898

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 2 .2148829 6.619704 5.25475 9.113658

RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 6 .3300716 1 .8641161

SLOPE = 5.82189 95 PERCENT CONFIDENCE LIMITS = 2.477106 AND 9.166672

INTERCEPT=-4.83532

LC50 = 6.769262 95 PERCENT CONFIDENCE LIMITS = 5.258215 AND 9.172377

LC25 = 5.184244 95 PERCENT CONFIDENCE LIMITS = 3.350522 AND 6.491309

LC10 = 4.077618 95 PERCENT CONFIDENCE LIMITS = 2.022414 AND 5.251234

LC05 = 3.531865 95 PERCENT CONFIDENCE LIMITS = 1.469189 AND 4.707148 *************************************************************************

M and E (1986) Fluridone Obs 16 TGAI 98 percent purity RBT 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 9 90 1.074219 5.6 10 5 50 62.30469 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 5.600001

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 2 .1677541 5.976884 4.838147 7.564668

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

48 RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 7 .3126901 1 .8491775

SLOPE = 6.831396 95 PERCENT CONFIDENCE LIMITS = 3.011368 AND 10.65142

INTERCEPT=-5.33119

LC50 = 6.031086 95 PERCENT CONFIDENCE LIMITS = 4.747503 AND 7.756306

LC25 = 4.804631 95 PERCENT CONFIDENCE LIMITS = 3.227234 AND 5.888167

LC10 = 3.915534 95 PERCENT CONFIDENCE LIMITS = 2.127572 AND 4.924097

LC05 = 3.464257 95 PERCENT CONFIDENCE LIMITS = 1.636563 AND 4.48257 *************************************************************************

M and E (1986) Fluridone Obs 16 TGAI 98 percent purity RBT 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 10 100 9.765625E-02 5.6 10 9 90 1.074219 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 5.6 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 4.490551

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 17 TGAI 98 percent purity RBT 24-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 10 100 9.765625E-02 5.6 10 2 20 5.46875 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

49 AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 6.721182

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS *************************************************************************

M and E (1986) Fluridone Obs 17 TGAI 98 percent purity RBT 48-hr LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 10 100 9.765625E-02 5.6 10 6 60.00001 37.69531 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 5.216388

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1906) Fluridone Obs 17 TGAI 98 percent purity RBT 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 10 100 9.765625E-02 5.6 10 8 80 5.46875 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 4.69561

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

50 M and E (1986) Floridone Obs 17 TGAI 98 percent purity RBT 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 10 100 9.765625E-02 5.6 10 10 100 9.765625E-02 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 5.6 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 4.233203

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 18 TGAI 98 percent purity RBT 48-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 7 70 17.1875 5.6 10 2 20 5.46875 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND +INFINITY CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 7.968173

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 1 .8025485 7.968173 5.367896 19.70378

RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 6 .4298382 1 .9863866

SLOPE = 5.966371 95 PERCENT CONFIDENCE LIMITS = 2.054696 AND 9.878046

INTERCEPT=-5.402087

LC50 = 8.043081 95 PERCENT CONFIDENCE LIMITS = 6.256024 AND 11.9153

LC25 = 6.199723 95 PERCENT CONFIDENCE LIMITS = 3.820752 AND 7.828876

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

51 LC10 = 4.904769 95 PERCENT CONFIDENCE LIMITS = 2.0888 AND 6.295384

LC05 = 4.263118 95 PERCENT CONFIDENCE LIMITS = 1.419552 AND 5.664675 *************************************************************************

M and E (1986) Fluridone Obs 18 TGAI 98 percent purity RBT 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 9 90 1.074219 5.6 10 4 40 37.69531 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 6.224119

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 2 .1677541 6.2839 5.114837 8.07079

RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 7 .3279173 1 .9512029

SLOPE = 7.139193 95 PERCENT CONFIDENCE LIMITS = 3.051001 AND 11.22739

INTERCEPT=-5.731041

LC50 = 6.349763 95 PERCENT CONFIDENCE LIMITS = 5.010802 AND 8.14885

LC25 = 5.108331 95 PERCENT CONFIDENCE LIMITS = 3.428347 AND 6.234012

LC10 = 4.199924 95 PERCENT CONFIDENCE LIMITS = 2.272586 AND 5.251398

LC05 = 3.73554 95 PERCENT CONFIDENCE LIMITS = 1.754007 AND 4.800982 *************************************************************************

M and E (1986) Fluridone Obs 18 TGAI 98 percent purity RBT 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 9 90 1.074219 5.6 10 6 60.00001 37.69531 3.2 10 0 0 9.765625E-02

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

52 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 5.216388

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 2 .1677541 5.684868 4.564247 7.109202

RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 7 .3025485 1 .6696461

SLOPE = 6.667483 95 PERCENT CONFIDENCE LIMITS = 3.000073 AND 10.33489

INTERCEPT=-5.055319

LC50 = 5.730664 95 PERCENT CONFIDENCE LIMITS = 4.504939 AND 7.370292

LC25 = 4.539859 95 PERCENT CONFIDENCE LIMITS = 3.053294 AND 5.575872

LC10 = 3.681192 95 PERCENT CONFIDENCE LIMITS = 2.009419 AND 4.644154

LC05 = 3.247134 95 PERCENT CONFIDENCE LIMITS = 1.544174 AND 4.217266 *************************************************************************

M and E (1986) Fluridone Obs 19 TGAI 98 percent purity RBT 48-h LC50 and Obs 21 RBT 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 9 90 1.074219 5.6 10 3 30 17.1875 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 6.711294

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 2 .1677541 6.618923 5.404147 8.654277

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

53 RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 7 .3472456 1 .990739

SLOPE = 7.6675 95 PERCENT CONFIDENCE LIMITS = 3.149231 AND 12.18577

INTERCEPT=-6.329082

LC50 = 6.690256 95 PERCENT CONFIDENCE LIMITS = 5.303757 AND 8.539605

LC25 = 5.463539 95 PERCENT CONFIDENCE LIMITS = 3.677296 AND 6.621589

LC10 = 4.55298 95 PERCENT CONFIDENCE LIMITS = 2.469681 AND 5.639651

LC05 = 4.082384 95 PERCENT CONFIDENCE LIMITS = 1.921691 AND 5.188412 *************************************************************************

M and E (1986) Fluridone Obs 19 TGAI 98 percent purity RBT 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 10 100 9.765625E-02 5.6 10 4 40 37.69531 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 6.027243

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 19 98 percent purity RBT 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 10 100 9.765625E-02 5.6 10 5 50 62.30469 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

54 CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 5.600001

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 20 TGAI 98 percent purity RBT 48-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 6 60.00001 37.69531 5.6 10 0 0 9.765625E-02 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND +INFINITY CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 9.291148

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 20 TGAI 98 percent purity RBT 72-h LC50 and Obs 22 RBT 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 9 90 1.074219 5.6 10 0 0 9.765625E-02 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 7.955181

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

55 M and E (1986) Fluridone Obs 20 TGAI 98 percent purity RBT 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 10 100 9.765625E-02 5.6 10 1 10 1.074219 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 7.039438

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 21 TGAI 98 percent purity RBT 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 6 60.00001 37.69531 5.6 10 2 20 5.46875 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 3.2 AND +INFINITY CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 8.699842

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 1 1.25215 8.699842 4.888956 +INFINITY

RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 6 .4923888 1 .9683006

SLOPE = 5.134785 95 PERCENT CONFIDENCE LIMITS = 1.531685 AND 8.737886

INTERCEPT=-4.826815

LC50 = 8.710094 95 PERCENT CONFIDENCE LIMITS = 6.668144 AND 16.01429

LC25 = 6.436709

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

56 95 PERCENT CONFIDENCE LIMITS = 3.802978 AND 8.527691

LC10 = 4.902653 95 PERCENT CONFIDENCE LIMITS = 1.721019 AND 6.446584

LC05 = 4.165608 95 PERCENT CONFIDENCE LIMITS = 1.025372 AND 5.694646 *************************************************************************

M and E (1986) Fluridone Obs 22 TGAI 98 percent purity RBT 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 8 80 5.46875 5.6 10 0 0 9.765625E-02 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND +INFINITY CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 8.331868

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 23 TGAI 98 percent purity FHM 24- ,48- and 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 32 10 10 100 9.765625E-02 18 10 0 0 9.765625E-02 10 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 18 AND 32 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 23.99999

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 23 TGAI 98 percent purity FHM 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT)

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

57 32 10 10 100 9.765625E-02 18 10 1 10 1.074219 10 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 18 AND 32 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 22.58704

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. ***********************************************************************

M and E (1986) Floridone Obs 24 EP 48 percent purity FHM 24-, 48-, 72-, and 96- h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10.2 10 10 100 9.765625E-02 6.7 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 6.7 AND 10.2 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 8.2668

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. ********************************************************************

M and E (1986) Fluridone Obs 25 TGAI 98 percent purity CCF 24-, 48-, 72- and 96-h LC50; (Obs 26 CCF 48-, 72-, and 96-h LC50; Obs 27 48-h LC50; Obs 28 24- and 48-h LC50; Obs 29 72- and 96-h LC50; Obs 30 72-h LC50; Obs 31 48-, 72-, and 96-h LC50; Obs 34 48-, 72-h, and 96-h LC50) (Obs 34) ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 10 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 10 AND 18 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 13.41641

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

58 NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. ************************************************************

Fluridone Obs 27 TGAI 98 percent purity CCF 24-h LC50 (Obs 29 48-h LC50) ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 9 90 1.074219 10 10 0 0 9.765625E-02 5.6 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 10 AND 18 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 14.27438

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 27 TGAI 98 percent purity CCF 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 10 10 2 20 5.46875 5.6 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 18 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 12.03225

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 27 TGAI 98 percent purity CCF 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 10 10 5 50 62.30469 5.6 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 18 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

59 AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 10

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. ************************************************************

Fluridone Obs 28 TGAI 98 percent purity CCF 72- and 96-h LC50 (Obs 30 96-h LC50) ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 10 10 1 10 1.074219 5.6 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 10 AND 18 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 12.61001

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. ************************************************************

M and E (1986) Fluridone Obs 32 TGAI 98 percent purity CCF 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 6 60.00001 37.69531 5.6 10 0 0 9.765625E-02 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02 .56 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND +INFINITY CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 9.291148

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

60 M and E (1986) Fluridone Obs 32 TGAI 98 percent purity CCF 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 10 10 9 90 1.074219 5.6 10 1 10 1.074219 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02 1 10 0 0 9.765625E-02 .56 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 7.483314

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 2 .1677541 7.483314 6.097914 10.30101

RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 7 .335756 1 .9999999

SLOPE = 10.19018 95 PERCENT CONFIDENCE LIMITS = 4.285535 AND 16.09482

INTERCEPT=-8.907669

LC50 = 7.484155 95 PERCENT CONFIDENCE LIMITS = 6.085822 AND 9.219152

LC25 = 6.426161 95 PERCENT CONFIDENCE LIMITS = 4.657015 AND 7.613862

LC10 = 5.602405 95 PERCENT CONFIDENCE LIMITS = 3.496446 AND 6.709696

LC05 = 5.160867 95 PERCENT CONFIDENCE LIMITS = 2.916097 AND 6.283215 *************************************************************************

M and E (1986) Fluridone Obs 33 TGAI 98 percent purity CCF 24-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 32 10 10 100 9.765625E-02 18 10 7 70 17.1875 10 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 10 AND 32 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 15.73982

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

61 WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 33 TGAI 98 percent purity CCF 48-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 32 10 10 100 9.765625E-02 18 10 8 80 5.46875 10 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 10 AND 32 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 14.95979

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 33 TGAI 98 percent purity CCF 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 10 10 6 60.00001 37.69531 5.6 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 18 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 9.291148

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. ************************************************************

M and E (1986) Fluridone Obs 34 TGAI 98 percent purity CCF 24-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 6 60.00001 37.69531 10 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 10 AND +INFINITY CAN BE

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

62 USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 16.70717

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 35 TGAI 98 percent purity BG 24 and 48-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 10 10 2 20 5.46875 5.6 10 1 10 1.074219 3.2 10 0 0 9.765625E-02 1.8 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 18 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 12.03225

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 2 .1677541 11.08247 8.965049 14.2681

RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 8 .3107925 1 .2334879

SLOPE = 6.497675 95 PERCENT CONFIDENCE LIMITS = 2.875301 AND 10.12005

INTERCEPT=-6.747158

LC50 = 10.92435 95 PERCENT CONFIDENCE LIMITS = 8.525309 AND 14.18352

LC25 = 8.6018 95 PERCENT CONFIDENCE LIMITS = 5.687832 AND 10.62462

LC10 = 6.936749 95 PERCENT CONFIDENCE LIMITS = 3.674683 AND 8.808607

LC05 = 6.09879 95 PERCENT CONFIDENCE LIMITS = 2.791151 AND 7.981655 *************************************************************************

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

63 M and E (1986) Fluridone Obs 35 TGAI 98 percent purity BG 72- and 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 10 10 2 20 5.46875 5.6 10 1 10 1.074219 3.2 10 0 0 9.765625E-02 1.8 10 1 10 1.074219 1 10 0 0 9.765625E-02 .56 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 5.6 AND 18 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 12.03225

RESULTS CALCULATED USING THE MOVING AVERAGE METHOD SPAN G LC50 95 PERCENT CONFIDENCE LIMITS 2 .1677541 11.08247 8.965049 14.2681

RESULTS CALCULATED USING THE PROBIT METHOD ITERATIONS G H GOODNESS OF FIT PROBABILITY 7 1.774067 3.911163 1.513898E-03

SINCE THE PROBABILITY IS LESS THAN 0.05, RESULTS CALCULATED USING THE PROBIT METHOD PROBABLY SHOULD NOT BE USED.

SLOPE = 3.173194 95 PERCENT CONFIDENCE LIMITS =-1.053313 AND 7.3997

INTERCEPT=-3.274289

LC50 = 10.76116 95 PERCENT CONFIDENCE LIMITS = 0 AND +INFINITY

LC25 = 6.596251 95 PERCENT CONFIDENCE LIMITS = 0 AND +INFINITY

LC10 = 4.245958 95 PERCENT CONFIDENCE LIMITS = 0 AND 10.23822

LC05 = 3.262011 95 PERCENT CONFIDENCE LIMITS = 0 AND 7.660581 ***************************************************************************

M and E (1986) Fluridone Obs 7-17-80 TGAI 98 percent purity CCF 72-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 13.5 10 10 100 9.765625E-02 10 10 7 70 17.1875 7.5 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 7.5 AND 13.5 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT

### Data Evaluation Record on the acute toxicity of Fluridone TGAI and EP to fish (rainbow trout, bluegill sunfish, channel catfish, and fathead minnow)

EPA MRID Number 00103297 and 40098001

64 CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 9.364391

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

M and E (1986) Fluridone Obs 7-17-80 TGAI 98 percent purity CCF 48-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 13.5 10 10 100 9.765625E-02 10 10 2 20 5.46875 7.5 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 7.5 AND 13.5 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 10.99063

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. *************************************************************************

Fluridone TGAI 98 percent purity 7-17-80 CCF 96-h LC50 ************************************************************************ CONC. NUMBER NUMBER PERCENT BINOMIAL EXPOSED DEAD DEAD PROB.(PERCENT) 18 10 10 100 9.765625E-02 13.5 10 10 100 9.765625E-02 10 10 10 100 9.765625E-02 7.5 10 0 0 9.765625E-02

THE BINOMIAL TEST SHOWS THAT 7.5 AND 10 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS, BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT.

AN APPROXIMATE LC50 FOR THIS SET OF DATA IS 8.660254

WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. **************************************************