1

OFFICE OF CHEMICAL SAFETY

AND POLLUTION PREVENTION

UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

WASHINGTON, D.C. 20460

 

Date: September 8, 2015

PC Code: 071003

MEMORANDUM DP Barcode: D427108

SUBJECT: Registration Review: Draft Problem Formulation for Environmental Fate, 

Ecological Risk, Endangered Species, and Human Health Drinking Water 

Exposure Assessments for Rotenone

TO: Kelly Ballard, Chemical Review Manager

Tom Moriarty, Team Leader

Neil Anderson, Branch Chief

Risk Management and Implementation Branch I

Pesticide Re-Evaluation Division (7508P)

FROM: Thomas Steeger, Ph.D., Senior Science Advisor

Cheryl Sutton, Ph.D., Environmental Scientist

Environmental Risk Branch IV

Environmental Fate and Effects Division (7507P)

REVIEWED

BY: Catherine Aubee, Senior Scientist

James Carleton, Ph.D., Senior Fate Scientist

Katrina White, Ph.D., Risk Assessment Process Leader

Environmental Risk Branch IV

Environmental Fate and Effects Division (7507P)

THROUGH: Jean Holmes, D.V.M., M.P.H., Branch Chief

Environmental Risk Branch IV

Environmental Fate and Effects Division (7507P)

1. Executive Summary

The Environmental Fate and Effects Division (EFED) has completed the preliminary problem 

formulation for the ecological risk, environmental fate, endangered species, and drinking water 

assessments to be conducted as part of the Registration Review of the piscicide rotenone (PC Code 

071003.; CAS No: 83-79-4). The problem formulation describes the methods to be used during the 

completion of drinking water exposure and ecological risk assessments in support of Registration 
2

Review and provides an overview of the environmental fate, ecological effects, and potential risks 

associated with the use of rotenone. It describes uncertainties which may be encountered in the 

risk assessment and identifies additional studies and other information that would be beneficial to 

its conduct. 

2. Introduction

Rotenone is an isoflavone pesticide used to kill fish (i.e., as a piscicide) in fisheries management. 

Although it was historically also used on cranberries; as an insecticide to control aphids, thrips, 

suckers, moths, and beetles in organic fruit and vegetable crops; as an insecticide/acaricide to 

control fire ants, for lice/tick control on pets; and for warble fly control on livestock, these uses 

have all been voluntarily cancelled by the technical registrants1

. Rotenone is a Restricted Use 

piscicide derived from the roots of tropical plants in the pea and bean family (Leguminosae) 

including jewel vine (Derris spp.) and lacepod (Lonchocarpus spp.) found in Australia, Oceania, 

southern Asia, and South America2

. The restricted use classification is based on the acute 

inhalation, acute oral and aquatic toxicity of the compound (USEPA 20073

). The mechanism of 

action has been well documented and it is understood that rotenone uncouples oxidative 

phosphorylation by blocking electron transport at complex I within the mitochondria. 

Rotenone is labeled for direct applications to mountain streams, lakes, ponds and fishery 

management operations. Rotenone has been used to restore Federally-listed threatened/endangered 

(listed) fish to their native habitats by eliminating competing fish. 

EFED examined the most recent environmental fate and ecological risk assessment,

4 written in 

support of the March 2007 Re-registration Eligibility Decision (RED) of rotenone,

5 in association 

with updated toxicity, exposure, and usage information to determine if additional data are needed 

to support the Registration Review of rotenone. In addition, EFED considered the latest Agency 

science policies and risk assessment methodologies. Since the RED, there has been an assessment 

to determine the risks from rotenone use to the federally threatened California red-legged frog 

(Rana aurora draytonii) as well as a FIFRA Section 18 emergency exemption petition6 submitted 

for the use of an unregistered formulation of rotenone as a piscicide.

 1 Cancellation orders for amendments to terminate uses: rotenone. http://www.regulations.gov/#!documentDetail;D=EPA-HQ-OPP2005-0494-0065 2 Finlayson, B. J., R. A. Schnick., R. L. Cailteux., L. DeMong., W. D. Horton , W. McClay , C. W. Thompson, and G. J. Tichacek.. 

2000. Rotenone Use in Fisheries Management: Administrative and Technical Guidelines Manual. American Fisheries Society, 

Bethesda, Maryland.

3 USEPA. 2007. Reregistration Eligibility Decision for Rotenone. Office of Chemical Safety and Pollution Prevention formerly the 

Office of Prevention, Pesticides and Toxic Substances (7509P). EPA 738-R-005. March 2007. Available on line at: 

http://www.epa.gov/oppsrrd1/reregistration/REDs/rotenone_red.pdf 4 USEPA. 2006. Environmental Fate and Ecological Risk Assessment Chapter (DP barcode D307382) n Support of Phase IV of the

Reregistration Eligibility Decision on Rotenone (PC Code 071003). Available on line at: 

http://www.regulations.gov/#!documentDetail;D=EPA-HQ-OPP-2005-0494-0031 5 Ibid USEPA. 2007. 6 USEPA. 2014. Review of Presidio Trust Section 18 Emergency Exemption Request for Use of CFT Legumine 3.3% (Rotenone) to 

Eradicate Invasive Fish in Mountain Lake, San Francisco, California (PC Code 071003; DP Barcode D421307). Dated July 15, 2013. 
3

3. Use Characterization

Rotenone has been used by fishery managers since the 1950s in the U.S. to kill fish in coves, 

shorelines, rivers and streams, and some estuarine habitats7

. Rotenone is typically applied to a 

surface water body as a piscicide in conjunction with a fisheries management or research program. 

Applicators must consult their local state Fish and Game Agency before applying rotenone 

products8

. In addition, the following use restrictions are typically recommended, although not 

required, when rotenone is applied to surface waters: (1) fish killed by using rotenone may not be 

consumed by people; (2) water treated with rotenone may not be used to irrigate crops; and (3) 

rotenone-treated water may not be released within one-half mile upstream of a drinking water 

supply intake or irrigation water intakes. Drinking water restrictions do not apply to municipalities 

that have an alternative potable raw water source that may be used until rotenone has dissipated 

from the treated water supply10.

Based on information from chemical profile on rotenone developed by the Biological and 

Economic Analysis Division (BEAD), rotenone is formulated as a dust, liquid, emulsifiable 

concentrate, and wettable powder (USEPA, 2015). Dust formulations must be mixed as a slurry to 

be applied to water surface by dragging equipment to release product behind a boat below the water 

surface. Applications can be made with aircraft, backpack sprayer, drip can, or handheld or handdirected nozzle to the water surface.

In lotic environments such as streams and rivers, rotenone is applied through drip stations or 

sprayers where the moving water carries the rotenone downstream. In lentic environments such as 

lakes, reservoirs and ponds, rotenone is typically pumped through a hose into the propeller wash of 

an outboard motor in order to distribute the chemical uniformly through the water column. 

Rotenone has been used extensively as a piscicide throughout the United States. By the mid-1980s, 

its greatest use was in the management of warm water fisheries in the Southeast and cool water 

fisheries of the Midwest and Mountain states (MRID 4558010-73). According to a survey 

conducted by the American Fisheries Society, during the period of 1988 to 2002 rotenone was used 

for fisheries management in 38 states and five Canadian provinces; eleven states and one Canadian 

province accounted for 89% of rotenone applications to surface waters in North America9

. 

Approximately 97% of rotenone applications during this period were made to standing waters 

(ponds and lakes). Reported uses were grouped into the following five categories, reflecting 

different fisheries management objectives: (1) obtain a sample of representative fish species and 

sizes to characterize fish populations (i.e., sampling; 34%); (2) manipulate fish populations to 

maintain desirable fish species for sport fisheries (e.g., reducing the density of planktivorous fish)

(27%); (3) treat rearing ponds (17%); (4) remove exotic species (10%); and (5) restore threatened 

and endangered species (7%). Formulated products used in lentic environments were roughly 

equally split between rotenone powder (53%) and liquid (47%)10.

Similar to other piscicides, rotenone is a restricted use product (RUP) and the chemical has 

relatively detailed standard operating procedures (SOPs), developed by the American Fisheries 

 7 Bettoli, P. W. and M. J. Maceina. 1996. Sampling with toxicants. Pages 303 - 333 in B.R. Murphy and D.W. Willis (editors), Fisheries 

Techniques, American Fisheries Society, Bethesda, MD.

8 U.S. EPA. 1988. Rotenone EPA Pesticide Fact Sheet 10/88. Fact Sheet Number 198. Available on-line at 

http://pmep.cce.cornell.edu/profiles/insect-mite/propetamphos-zetacyperm/rotenone/insect-prof-rotenone.html

9 McClay W. 2005. Rotenone use in North America (1988-2002). Fisheries. 30(4): 29-31.

10 Ibid McClay 2005.
4

Society, and associated with its use to renovate/restore fish populations11. Since the SOPs are 

linked to (i.e., specified on) the federal label, they are considered part of the label and must be 

followed by the end user. The current SOP (USDOI 2010) states that, “The flow of a stream or 

outflow of a treated lake beyond the treatment area now requires chemical deactivation with 

potassium permanganate unless it is demonstrated to be unnecessary.” An example of a situation 

in which deactivation would not be conducted is when it is deemed unfeasible, such as when the 

stream flow goes into a canyon (or other physical limitation) or “if there is no discharge from the 

Treatment Area or the discharge goes dry in a distance shorter than 2 miles or 2 hours travel-time 

(maximum distance/time between drip stations recommended on label) from the lowest drip 

station.” For the first scenario, “the Certified Applicator through bioassay or analytical testing 

assures that the discharge is no longer toxic at 30 minutes travel time downstream of where the 

stream emerges at an accessible location and deactivation could be accomplished.” Examples of 

the latter scenario include ponds or lakes with no discharge or a stream that goes dry within two 

miles of the treatment area. An additional scenario in which deactivation is not required is when 

dilution of untreated waters renders “undetectable concentrations” (i.e., <2 ppb) in discharge 

waters. Regardless of whether the compound is deactivated, bioassays are typically conducted and 

cages containing sentinelspecies (typically the target fish species) are placed downstream of treated 

waters to ensure that the chemical does not substantially move outside of the targeted treatment 

area. 

 11 Finlayson, B. J., R. A. Schnick., D. Skaar, J. Anderson, L. Demong, D. Duffield, W. Horton, and J. Steinkjer. Planning and Standard 

Operating Procedures for the Use of Rotenone in Fish Management. Rotenone SOP Manual. American Fisheries Society Fish 

Management Chemicals Subcommittee. American Fisheries Society, Bethesda, Maryland. Available on line at: 

http://www.fisheriessociety.org/rotenone/rot.pdf
5

4. Conclusions from Previous Risk Assessments

The ecological risk assessment written in support of the RED document for rotenone was issued in 

September 200612. Based on the previous analysis of the environmental fate, ecotoxicity, and 

registered uses of rotenone, exposure to the compound resulting from the maximum treatment 

concentration is expected to kill fish, aquatic-phase amphibians, and aquatic invertebrates in the 

immediate treatment area. However, in lotic environments the compound is typically deactivated

with a strong oxidizing agent (e.g., potassium permanganate; KMnO4) to ensure that it does not 

move outside of the intended treatment area. Based on the most sensitive species and at the 

maximum treatment concentration, risk quotient (RQ) values exceed acute risk levels of concern 

(LOCs) by several orders of magnitude. 

While the conclusions presented below represent the most recent assessments, there have been 

changes made in the SOP that will affect the new assessments. The major changes are:

• Maximum Rotenone Treatment Levels—The maximum treatment level in standing waters 

was reduced from 250 to 200 ppb rotenone (a.i.), and the maximum treatment level in 

flowing water was increased to 200 ppb rotenone (a.i.). For all applications, the selected 

treatment rate is based on response of target fish (or surrogate species) in a bioassay with 

site water (or in similar water) within the maximum level on the label (SOP 5).

• Chemical Deactivation of Treated Flowing Water—The flow of a stream or outflow of a 

treated lake beyond the treatment area now requires chemical deactivation with potassium 

permanganate unless it is demonstrated to be unnecessary (SOP 7). The deactivation zone 

and other areas affected by the treatment are included in the definition of a project area 

(SOP 6).

a. Ecological Risk Assessment

The major environmental fate and ecological risk conclusions presented in ecological risk 

assessments13 in support of the RED (when the maximum treatment concentration was 250 μg/L, 

rather than the current 200 μg/L) were as follows:

The use of rotenone as a piscicide is intended to kill target species. The chemical’s use at a 

treatment site is intended to kill fish and at maximum application rates (250 μg/L), exceeds the 

acute risk level of concern by factors of 206X and 108X for freshwater fish and invertebrates, 

respectively. Maximum treatment concentrations exceed the upper 95th percentile of 96-hr median 

lethal concentrations for freshwater fish species sensitivity distribution. The extent of acute 

mortality that will be inflicted on aquatic animals within a treatment area is uncertain; however, 

acute effects are expected. At the maximum application rate in cold water, the chronic risk level of 

concern is exceeded for freshwater fish and invertebrates by factors of 175X and 153X, 

respectively. To the extent that lower application rates are used and that less sensitive species are 

present than those used to evaluate risk in this assessment, acute mortality among non-target aquatic 

animals may be substantially lower. 

Benthic macroinvertebrate monitoring data indicate that repeated yearly rotenone treatments 

conducted in mountain streams in California did not substantially reduce aquatic invertebrate 

abundance and that aquatic communities tended to return to pretreatment conditions one to two 

 12 Ibid USEPA 2006. 13 Ibid USEPA 2006.
6

years after treatment; whether these aquatic communities achieved species diversity similar to 

pretreatment conditions is uncertain. The likelihood of chronic exposure following the treatment 

of streams is considered low since rotenone is typically applied only once per year and the chemical 

is continually diluted via displacement by inflowing water and, and by abiotic degradation; 

however, is lentic systems where rotenone is not intentionally deactivated and where only a portion 

of the water body has been treated, there is potential for chronic exposure. The extent to which 

rotenone’s use as a piscicide will affect aquatic animals outside of the treatment area depends on 

the extent to which the chemical moves beyond the treatment area. Although not required by the 

label, relatively rigorous application and monitoring efforts may be conducted by resource 

managers using this compound and standard operating procedures typically recommend that 

rotenone be deactivated with potassium permanganate when there is a likelihood that the piscicide 

will move outside of the treatment area (e.g., see SOP 7 on Determining Need and Methods for 

Chemically Induced Deactivation14). Additionally, rotenone is a restricted use pesticide when used 

to kill fish. These procedures and practices would reduce the risk of non-target aquatic animal 

mortality by limiting the movement of rotenone outside of treatment areas to the extent possible. 

The use of potassium permanganate to deactivate rotenone can result in acute toxicity as well for 

aquatic organisms which are in close proximity to the deactivation station. A more detailed analysis 

of the risk associated with potassium permanganate alone is contained in the 2006 risk assessment15

and the reader is referred to that assessment. 

Since rotenone is applied directly to water, the risk of terrestrial animal acute mortality is 

considered low since there are not likely to be any rotenone residues on terrestrial animal forage 

items. Although no specific application protocol is required, in cases where application protocols 

require that dead and/or dying fish be collected and buried, there is reduced opportunity for either 

birds or mammals to consume fish that may contain rotenone residues. However, even if birds or 

mammals were to consume fish killed by rotenone, there would be insufficient quantities of 

rotenone in the carcasses to represent a risk of acute mortality in terrestrial animals. Since fish are 

either collected or shortly sink to the bottom of treated water and rapidly decompose, the likelihood 

of chronic exposure through the diet of terrestrial animals is also considered to be low. 

In addition, effects on threatened and endangered species may be an important consideration for 

site-specific applications that are co-located with critical habitat. Rotenone is an important 

chemical used in recovery efforts for endangered salmonid species by eliminating competing 

species. Monitoring may be conducted by resource managers prior to and after the use of rotenone 

as a piscicide. It is not considered likely that these resource managers would disregard the presence 

of an endangered/threatened species in a projected treatment area. 

Previously, acute and chronic risk to estuarine/marine fish and invertebrates could not be 

determined because toxicity data were not available, and the possibility of risk from acute or 

chronic exposure to these receptors could not be precluded. However, the RED16 stipulated that 

rotenone labels had to be updated to prohibit use in estuarine/marine environments. Also, an 

evaluation of risk to aquatic plants could not be conducted since there were no pertinent toxicity 

data and the possibility of risk from exposure in treated waters could not be precluded.

 14 American Fisheries Society. 2010. Determining Need and Methods for Chemically Induced Deactivation SOP 7 In Finlayson et al. 

Planning and Standard Operating Procedures for the Use of Rotenone in Fish Management. Rotenone SOP Manual. American 

Fisheries Society Fish Management Chemicals Subcommittee. http://www.afsbooks.org/55061P 15 Ibid USEPA 2006. 16 Ibid USEPA. 2007
7

b. Drinking Water Exposure Assessments

EFED conducted a drinking water assessment of rotenone17 in 2005 as part of the RED process. 

The most conservative acute concentrations were considered as the maximum treatment 

concentrations, at the time of the assessment, of 250 μg/L and 50 μg/L for lakes and rivers, 

respectively. Chronic exposure values were not estimated for the piscicidal uses given that the 

compound was intentionally deactivated in some environments. However, based on the previously 

registered agricultural uses, which have subsequently been cancelled, the peak, annual mean and 

overall mean drinking water exposure concentrations were 1.83 and 0.0049 and 0.0028 μg/L, 

respectively18.

c. Clean Water Act Programs 

Rotenone is not identified as a cause of impairment for any water bodies listed as impaired under 

section 303(d) of the Clean Water Act. No Total Maximum Daily Loads (TMDLs) or water quality 

criteria have been developed for rotenone.

19

5. Environmental Fate and Transport

Rotenone or (6R, 6aS, 12aS)-1,2,6,6a,12,12a-hexahydro-2-isopropenyl-8,9-

dimethoxychromenyl[3,4-bfuro[2,3-h]chromen-6-one, is a biochemical related to and derived 

from isoflavonoid compounds by plants. Rotenone has three chiral centers, and only one of the 

eight possible stereoisomers is actually produced by plants. As with other stereoisomers, the 

environmental fate and biological effects can be influenced by the relative positions of atoms 

around chiral centers.

The physical and chemical properties that determine the fate and transport of rotenone are detailed 

in Table 1. The environmental fate data set for rotenone is incomplete, as the majority of studies 

submitted previously do not meet the current standards for environmental fate guideline data 

requirements (OCSPP Series 835). Based on available data, rotenone has a relatively low potential 

to bioconcentrate in aquatic organisms (BCF <30). Based on its estimated low vapor pressure and 

estimated low Henry’s Law constant, rotenone is not expected to volatilize. 

When applied to water, rotenone is expected to hydrolyze, but data are required to accurately

determine the rates at which this will occur at various environmental pH levels, and to characterize

the transformation products. Based on information available in the literature, aqueous photolysis 

may also occur, but acceptable guideline data have not been submitted to address this degradation 

route. The extent of degradation through biotic mechanisms is unknown since no acceptable

metabolism data are available to assess this potential route of degradation. Rotenone is expected 

to partition somewhat to the sediment phase of an aquatic environment, although the extent to 

which that will occur is unclear, as the submitted adsorption/desorption data do not meet guideline 

requirements. 

Although an acceptable guideline aquatic field dissipation study has not been submitted, 

supplemental information is available through submitted studies on the aquatic dissipation of 

 17 USEPA. 2005. Drinking Water Exposure Assessment for Rotenone. DP Barcode D320702. Dated September 15, 2005. Available 

on line at: http://www.regulations.gov/#!documentDetail;D=EPA-HQ-OPP-2005-0494-0009 18 Ibid USEPA. 2005. 19 USEPA. 2015. Watershed Assessment, Tracking and Environmental Results. Specific State Causes of Impairment that Make Up 

the National Pesticides Cause of Impairment Group.

http://iaspub.epa.gov/tmdl_waters10/attains_nation_cy.cause_detail_303d?p_cause_group_id=885 \
8

rotenone (at 250 ppb) in two experimental ponds in Wisconsin that were maintained at two different 

temperatures (5o

C and 23–27oC). Rotenone dissipated from the water column with half-lives of 23 

hrs in the cold water pond and 10.6 hours in the warm-water pond (Acc. # 00157851/TRID 

4701520-10). Rotenone degraded from the entire system (water + sediment) with half-lives of 

approximately 20 days in the cold-water pond and 1.5 days in the warm-water pond. Only the parent 

was assessed in this study. As would be expected, all the resident fish in the ponds died shortly 

after the initial application. After 7 days in the cold-water pond and 1 day in the warm-water pond, 

fat-head minnows (Pimephales promelas) were placed in the pond in cages until 9 of 10 survived 

for 24 hours. This occurred after 4 days in the warm pond and 30 days in the cold-water pond. 

Notably, the pH near 8.5 in both ponds would have been expected to result in fairly rapid hydrolysis 

of rotenone based on available information. While this occurred in the warm-water pond, it did not

in the cold water pond, suggesting that rotenone degradation may slow substantially in cooler

environments.

Much of the concentration data related to the application of rotenone to California’s Lake Davis to 

control northern pike (Esox lucius) was collected in 1997 (California Department of Fish and Game, 

1999). Lake Davis, which is in the Plumas National Forest, has a surface area of 4030 acres and a 

normal capacity of 84,370 acre-feet. The dataset can be used to estimate the dissipation rate of 

rotenone from Lake Davis after the initial application, which was made on October 17, 1997 (rather 

late in the season for rotenone application) when the water was fairly cold (9o

C). Measurements 

were made at nine sites on each of six time periods extending out to 25 d post-application. The 

mean concentration of rotenone in the lake on the day of application was 45 μg∙L-1

. A control 

sample taken prior to application showed that no rotenone or rotenolone was present. Rotenone 

dissipated from the reservoir with a half-life of 10.3 d (USEPA, 2008). Approximately 55% of the 

rotenone formed rotenolone. Rotenolone dissipated with a half-life of 5.5 d. Some portion of the 

dissipation of both rotenone and rotenolone would have been due to discharge from the lake; the 

volume of discharge was not reported.

Table 1. Chemical Properties and Environmental Fate Parameters of Rotenone.

Chemical/Fate Parameter Value Description/Comments Source (MRID)

Selected Physical/Chemical Parameters

Chemical name

(6R, 6aS, 12aS)-

1,2,6,6a,12,12a-hexahydro-2-

isopropenyl-8,9-

dimethoxychromenyl[3,4-

bfuro[2,3-h]chromen-6-one

 ̄  ̄

Rotenone CAS Number 83-79-4  ̄  ̄

SMILES Notation COc5cc4OCC3Oc2c1CC(Oc1c

cc2C(=O)C3c4cc5OC)C(C)=C  ̄  ̄

Molecular formula C23H22O6  ̄  ̄

Molecular mass 394.4 g mol-1  ̄ Tomlin, 199420

Vapor pressure (25°C) 6.9 x 10-10 Torr Not likely to volatilize. EPISuiteTM v4.11 

Water solubility (20°C) 0.2 mg/L  ̄ Augustijn-Beckers, 

199421

 20 Tomlin, C. 1994. Pesticide Manual 10th edition. British Crop Protection Council. The Royal Society of Chemistry, Cambridge U.K. 21 Augustijn-Beckers P, A. G. Hornsby, and R. D Wauchope. 1994. The SCS/ARC/CES pesticide properties database for environmental 

decision-making. Additional compounds. Rev. Environ. Contam. Toxicol. 137:1-82.
9

Chemical/Fate Parameter Value Description/Comments Source (MRID)

Octanol-water partition 

coefficient (Log KOW) 4.10  ̄ Hansch et al., 199522

Henry’s Law Constant 

(25°C) 1.1x10-13 atm-m3 mol-1 Not likely to volatilize from moist soil or water. EPISuiteTM v4.11

Persistence

Hydrolysis half-life (days;

25°C) No data.  ̄  ̄

Aqueous photolysis half-life 

(25°C) No data.  ̄  ̄

Soil photolysis half-life 

(25°C) No data. Not required for aquatic 

use. ˗

Aerobic soil metabolism 

half-life (25°C) No data.  ̄  ̄

Anaerobic soil metabolism 

half-life (25°C) No data. Not required for aquatic use. ˗

Aerobic aquatic metabolism 

half-life (25°C) No data.  ̄  ̄

Anaerobic Aquatic half-life 

(25°C) No data.  ̄  ̄

Mobility

Soil-water partition 

coefficients (Kd); No data.  ̄  ̄

Organic carbon-normalized 

soil partition coefficients 

(KFOC)

No data.  ̄  ̄

Field Dissipation

Aquatic field dissipation 

half-life

1.5 days (5 °C)

20 days (23-27 °C)

10.3 days (parent)

5.5 days (rotenolone)

 ̄

Acc. # 00157851

(TRID 470152-010)

MRID 42217701

Fish Bioconcentration

Fish bioconcentration factors 

(steady-state) 

10.8X (viscera)

27.9X (head)

27.6X(carcass)

 ̄ USEPA 2008

(Acc.#’s

146183, 143252)

 22 Hansch, C., A. Leo, and D. Hoekmann. 1995. Exploring QSAR. Hydrophobic, Electronic, and Steric Constants. ACS Prof Ref Book. 

Heller SR, consult. ed., Washington, DC: Amer Chem Soc.Hazardous Substances Data Bank (HSDB). 2004. Available through 

TOXNET, NLM. http://toxnet.nlm.nih.gov/.
10

a. Transformation Products

The major degradate of rotenone, previously identified in submitted studies, is 6αß, 12 αßrotenolone, which is structurally similar to the parent compound. Identification of additional 

degradates may occur once acceptable environmental fate data are submitted.

6. Receptors

The most sensitive aquatic and terrestrial toxicity data for rotenone are summarized in Tables 2

and 3, respectively. Within each of these very broad taxonomic groups, an acute and/or chronic 

endpoint is selected from the available test data and is used as a denominator in ratios comparing 

point estimates of exposure to point estimates of effect. 

Table 1. Rotenone toxicity values (μg a.i./L) for aquatic organisms.

Species

Toxicity

Value

(μg a.i./L)

Effects 

Endpoint

Exposure 

Duration

Toxicity 

Classification

Reference 

(classification)

Acute toxicity

Freshwater Fish 

Rainbow Trout

(Oncorhynchus mykiss)

LC50 = 1.94

98% a.i.

Survival 96-hour Very highly 

toxic

MRID 439751-0223

(Acceptable)

Freshwater 

Invertebrates Daphnid

(Daphnia magna)

EC50 = 3.7

96.5% a.i.

Survival 48-hour Very highly 

toxic

MRID 400633-0324

(Supplemental)

Estuarine/Marine Fish No data

Estuarine/Marine 

Invertebrates No data

Aquatic Plants No data

Chronic Toxicity

Freshwater Fish 

Rainbow Trout

(Oncorhynchus mykiss)

NOAEC = 1.01

96.5% a.i.

Growth 32-day NR

MRID 400633-0225

(Supplemental)

Freshwater 

Invertebrates Daphnid

(Daphnia magna)

NOAEC = 1.25

96.5% a.i. Reproduction 21-day NR

MRID 400633-0326

(Supplemental)

 23 Sousa, J. 1996. Crystalline Rotenone--Acute Toxicity to Rainbow Trout (Oncorhynchus mykiss) Under Flow-Through Conditions: 

Lab Project Number: 10824.0625.6119.108: 95-10-6154: 051995/FIFRA/108. Unpublished study prepared by Springborn Laboratories, 

Inc. 70 p. MRID 439751-01.

24 Rach, J., T. Bills, and L. Marking. 1986. Rotenone--Acute LC50 Freshwater Invertebrate--Daphnia: TOX 83-626/01A. Unpublished 

study prepared by U.S. Fish and Wildlife Service. 26 p. MRID 400633-03. 25 Bills, T., J. Rach, and L. Marking. 1986. Rotenone--Fish Early Life Stage--Rainbow Trout: Laboratory Project ID; TOX 83-626.01B. 

Unpublished study prepared by U.S. Fish and Wildlife Service. 15 p. MRID 400633-02. 26Rach, J. J., T. D. Bills, and L. L. Marking. 1986. Rotenone –aquatic invertebrate life cycle—Daphnia. U.S. Fish and Wildlife Service, 
11

Species

Toxicity

Value

(μg a.i./L)

Effects 

Endpoint

Exposure 

Duration

Toxicity 

Classification

Reference 

(classification)

Estuarine/Marine Fish No data

Estuarine/Marine 

Invertebrates

No data

NR = not relevant; EFED has not established toxicity classifications for chronic endpoints.

Table 2. Rotenone toxicity values for terrestrial organisms.

Species % a.i. Toxicity Value Toxicity 

Category Reference

Study 

Classification

Mammalian Acute Oral Toxicity

Rat

(Rattus norvegicus)

99.2%

LD50:

102 mg a.i./kg 

(Males)

39.5 mg a.i./kg 

(Females)

Highly toxic Acc. 0014549627 Acceptable

Mammalian Chronic Toxicity

Rat

(Rattus norvegicus)

Two-generation 

reproductive study

97.9%

NOAEL = 7.5 

mg/kg (0.5 and 

0.6 mg/kg/day 

for males and 

females, 

respectively) 

based on 

decreased body 

weight and body 

weight gain

NA Acc. 0014140828 Acceptable

Avian Acute Oral Studies

Mallard duck

(Anas platyrhynchos)

32.4% LD50 = 2200 

mg/kg bw

Practically 

nontoxic Acc. 14325029 Supplemental

Ring-necked pheasant

(Phasianus colchicus) 32.4 LD50 = 1680 

mg/kg bw

Slightly 

toxic MRID 14325030 Supplemental

Avian Subacute Dietary Studies

 National Fisheries Research Center, LaCrosse, Wisconsin

27 Eiseman, J. and A. Thakur. 1984. General Metabolism Study for Safety Evaluation of Rotenone using Rats. Project No. 419-137. 

Unpublished Study Prepared by Hazleton Laboratories, America, Inc. 311 p. Accession number: 000145496.

28 Kehoe, D, K. MacKenzie. 1983. Reproduction Study for Safety Evaluation of Rotenone Using Rats. Final Report. Study No 

81077. Unpublished study prepared by Hazleton Raltech, Inc. 732p. Accession number: 000141408. 29 Tucker, R. 1968. Rotenone: [Acute Oral Toxicity to Mallard Ducks and Ring-necked Pheasants]. Unpublished study prepared by 

U.S. Fish and Wildlife Service, Denver Wildlife Research Center. 8 p. Accession number: 000143250.

30 Ibid Tucker 1968.
12

Species % a.i. Toxicity Value Toxicity 

Category Reference

Study 

Classification

Mallard duck

(Anas platyrhynchos) 34.5

5-day LC50:

2600 mg/kg diet

Slightly 

toxic Acc. No. 24878831 Supplemental

Ring-necked pheasant

(Phasianus colchicus)

34.5

5-day LC50:

1608 mg/kg diet

Slightly 

toxic Acc. No. 24878832 Supplemental

Japanese quail

(Coturnix japonica)

34.5

5-day LC50:

1882 mg/kg diet

Slightly 

toxic Acc. No. 24878833 Supplemental

Honey bee Acute Contact Toxicity

Honeybee

(Apis mellifera)

> 95

LD50:

> 60 μg a.i./bee

Practically 

non-toxic

Acc. No.

0500199134 Acceptable

Honeybee

(Apis mellifera)/

48 hour

Technical

2.4 μg a.i./bee 

elicited 12% 

mortality

Moderately 

toxic

Acc. No. 

0003693535 Supplemental 

a. Ecological Incidents

The Ecological Incident Information System (EIIS) is an internal OPP database that houses 

ecological incidents that have been reported to the Agency. When available, EIIS includes date 

and location of an incident, type and magnitude of effects observed in various species, use(s) of 

pesticides known or suspected of contributing to the incident, and results of any chemical residue 

analysis or other analyses conducted during incident investigation. EIIS incidents are categorized 

according to the relative certainty that the incident resulted from pesticide exposure. A search of 

the EIIS for ecological incidents run on February 19, 2015, identified six incident reports which 

were reported over the period from 1977 to 1999. Three of the incidents were classified as having 

occurred as a result of misuse of the compound while for the remaining three the legality of use 

was undetermined. All but one of the reported incidents involved the loss of fish; however, one 

incident reported the loss of swans and geese. Since the incident occurred several days after the 

rotenone was applied, the report indicated that it was “unlikely” that the loss was associated with 

the rotenone application. Incidents may have occurred due to rotenone exposures but may not have 

been reported due to various factors, such as a lack of reporting, or a lack of observation of effects.

Therefore, the lack of incident reports does not necessarily indicate the absence of incidents; 

however, the restricted use status of the compound coupled with its detailed SOPs may limit the 

extent of unintended exposure. 

 31 Hill, E. F., R. G. Heath, J. W. Spann and J. D. Williams. 1975. Lethal dietary toxicities of environmental pollutants to birds. U. S. 

Fish and Wildlife Service, Special Scientific Report Wildlife No. 191. Washington, D. C. Accession number: 000248788. 32 Ibid Hill et al. 1975. 33 Ibid Hill et al. 1975. 34 Stevenson, J. H. 1978. The Acute Toxicity of Unformulated Pesticides to Worker Honey Bees (Apis mellifera L.). Plant Pathology 

27(1): 38 - 40. Accession number: 005001991.

35 Atkins, E. L., E. A. Greywood and R. L. Macdonald. 1975. Toxicity of Pesticides and Other Agricultural Chemicals to Honey 

Bees: Laboratory Studies. University of California, Department of Entomology, U.C. Cooperative Extension (Leaflet 2287) 38p. 

Accession number: 00036935.
13

No plant or wildlife incidents have been reported through OPP’s aggregate incident report database 

and no avian incidents have been captured in the Avian Incident Monitoring System (AIMS; 

American Bird Conservancy 2013)36 as of February 10, 2015. The AIMS database contains 

publicly available reports on pesticide-related avian incidents through 2005.

7. Exposure Pathways of Concern

For a pesticide to pose an ecological risk, it must reach ecological receptors in biologically 

significant concentrations. An exposure pathway is the means by which a pesticide moves in the 

environment from a source to an ecological receptor. For an ecological pathway to be complete, it 

must include a source, a release mechanism, an environmental transport medium, a point of 

exposure for ecological receptors, and a feasible route of exposure.

When used as a piscicide, rotenone is applied directly into, or onto the surface of, a receiving water

body, which may be a stream, pond, or lake/reservoir. Once in the water column, non-target aquatic 

organisms may be exposed to rotenone via three main exposure pathways and two minor pathways. 

1. Rotenone will initially be distributed in the water column of the receiving water, and 

organisms in the water column (aquatic invertebrates, fish, amphibians and plants) may be 

directly exposed through contact. Organisms can take up soluble rotenone through the gills 

or integument. This is expected to be their main route of exposure to rotenone. If sufficient 

concentrations exist, then individual toxic effects in aquatic invertebrates, fish, aquaticphase amphibians and/or plants can result. This may result in reductions in populations of 

species within these taxa.

2. Rotenone released into water will likely sorb to sediment and suspended solids. Receptors 

such as aquatic invertebrates, fish, and aquatic-phase amphibians may be exposed to sorbed 

rotenone through incidental ingestion of or contact with sediment and/or suspended solids. 

This can result in changes in population size from reduced survival, growth and 

reproduction.

3. Rotenone may sorb to plant surfaces. Ingestion of plant materials with sorbed rotenone

can result in a similar exposure pathway as described for sediment and suspended solids. 

Nonvascular or vascular plants may also be exposed to rotenone through direct contact or 

root uptake. This could result in changes in plant populations due to direct toxicity. 

However, since no plant toxicity data are available for rotenone, potential risks to aquatic 

plants cannot be evaluated. 

4. Rotenone residues may accumulate in target and non-target organisms, although this is not 

considered likely given the low estimated bioconcentration factor. Other organisms that 

live in or around the treatment area may be exposed to rotenone by consuming treated 

organisms.

5. Direct exposure of terrestrial animals/plants to rotenone is not likely since the chemical is 

only applied to water bodies. Indirect exposure to both mammals and birds is possible 

through the consumption of dead and dying fish and larger invertebrates and through 

 36 Available at: http://www.abcbirds.org/abcprograms/policy/toxins/aims/aims/index.cfm
14

ingestion of rotenone-treated water or through ingestion of plants or aquatic animals in 

treated areas. Given that aerial application are permitted on some labels, spray drift may 

also serve as a potential route of exposure.

Environmental fate data suggest (but have not clearly demonstrated) that rotenone has low potential 

to volatilize; thus, the likelihood of inhalation exposure is considered low. The compound is applied 

directly to water; therefore, the opportunity for inhalation exposure in birds is considered low.

Potential exposure through ingestion of rotenone-treated water was evaluated using the Screening 

Imbibition Program (SIP; version 1.0). The model is intended to provide an upper bound of 

exposure to birds and mammals to pesticides through drinking water alone. Results from the 

Excel®-based tool are provided in Appendix C and indicate that drinking water exposure alone is 

not a potential concern for acute risk to birds or mammals. The available data also indicate that 

exposure through drinking water is not a potential concern for chronic risk to mammals; however, 

since no chronic toxicity data are available for birds, there are insufficient data to preclude chronic 

risk.

8. Analysis Plan

In order to address the risk hypothesis, the potential for adverse effects on the environment is 

estimated. The uses, environmental fate, and ecological effects of rotenone are characterized and 

integrated to assess the risks. This is accomplished using a risk quotient (i.e., ratio of exposure 

concentration to effects concentration) approach. Although risk is often defined as the likelihood 

and magnitude of adverse ecological effects, the risk quotient-based approach does not provide a 

quantitative estimate of likelihood and/or magnitude of an adverse effect.

This analysis plan may be revised depending upon the information submitted by the public in 

response to the opening of the Registration Review docket for rotenone.

a. Stressors of Concern

i. Ecological Risk Assessment

The stressors of concern are as follows:

• Terrestrial and Aquatic Environments: rotenone (parent only) at this time. However, 

submission of new data which identifies and quantifies environmental degradates may 

lead to determination of additional compounds which will be included as stressors of 

concern in the ecological exposure estimates. These data will be considered along with 

the requirement for deactivation of the parent compound in flowing waters or standing 

waters that have a means of discharge.

ii. Drinking Water 

A new drinking water assessment (DWA) will be conducted for the Registration Review of 

rotenone to incorporate the changes to the maximum treatment concentrations as stated in the new 

SOPs (USDOI 2010). EFED conducted a drinking water assessment for rotenone in 2005 in which 

the most conservative acute concentration was considered the maximum treatment concentration 
15

of 250 μg/L and 50 μg/L for lakes and rivers, respectively. These values, which are target treatment 

concentrations and not results from modeling, would decrease as the treatment concentrations for 

rotenone have changed. The current maximum treatment concentration for all bodies of water 

(flowing and standing) is 200 μg/L. Chronic exposure values were not estimated in the 2005 DWA

for the piscicidal uses (the only remaining uses currently on the label) given that the compound was

intentionally deactivated in some environments. Additionally, the compound is expected to 

degrade via abiotic processes. Because the new SOP requires deactivation, chronic exposure values 

will not be determined in the new DWA. 

b. Measures of Exposure 

The registered use of rotenone as a piscicide requires the direct application of the compound to 

water to maintain specific water concentrations for defined durations. Therefore, aquatic exposure 

numbers are specific to meet the management objective of the treatment. As such, exposure 

modeling per se is not needed for rotenone as actual treatment levels will be used for aquatic 

estimated environmental concentrations (EEC).

i. Available Monitoring Data

The Agency is not aware of surface water or groundwater monitoring conducted by federal or state 

agencies; monitoring within the treatment area is typically conducted via bioassay, i.e., caged live 

fish. However, if monitoring data of rotenone become available, they will be considered in the 

assessment.

c. Measures of Effect 

Toxicity data presented in Section 3 of this problem formulation will be used to calculate risk 

quotients. Any additional information submitted by the registrant or found in the open literature 

meeting the standards established by the Agency prior to conduct of the risk assessment will also 

be considered. The open literature studies are identified using EPA’s ECOTOXicology database 

(ECOTOX)37, which employs a literature search engine for locating chemical toxicity data for 

aquatic life, terrestrial plants, and wildlife. The evaluation of both sources of data can also provide 

insight into direct and indirect effects of pesticides on biotic communities from loss of species that 

are sensitive to the chemicals and from changes in structure and functional characteristics of the 

affected communities.

9. Endangered Species Assessments

Consistent with the Agency’s responsibility under the Endangered Species Act (ESA), the Agency 

will evaluate risks to listed species from registered uses of pesticides in Registration Review. The 

process for evaluating potential risks to listed species is further described at 

http://www.epa.gov/oppfead1/endanger/litstatus/riskasses.htm.

 37 http://cfpub.epa.gov/ecotox/
16

10. Endocrine Disruptor Screening Program (EDSP)

Rotenone was not included in the first group of 67 chemicals issued an order to conduct Tier 1 and 

Tier II EDSP testing. For additional information the EDSP program, visit 

http://www.epa.gov/endo/. 

11.Preliminary Identification of Data Gaps

a. Environmental Fate

Table 4 identifies environmental fate studies that were submitted for each guideline requirement, 

as well as current study classifications and whether or not further data are needed in order to support 

risk assessment. Environmental fate data that were previously submitted were reviewed using 

current standards. This review resulted in several studies being classified as “not acceptable” due 

to various deficiencies in the experimental method and/or the analytical method, as well as 

additional deficiencies related to OCSPP Series 835 guideline study requirements.

In the 1983 hydrolysis study, transformation products were inadequately characterized. 

Unidentified residues in the organic and aqueous extracts were present at up to 48% and 16% of 

the applied, respectively. Unidentified residues at TLC plate origins were present at up to 12% of 

the applied. Material balances were incomplete, with losses of 26 - 48% of the applied across all 

three pH levels studied. Additionally, the concentration of the co-solvent was not reported and it 

was not confirmed that the test vessels were sterilized. 

The 1982 aqueous photolysis study was not conducted at the environmental pH level at which 

hydrolysis was expected to be slowest (i.e., pH 5), but was studied at pH 7.6. The compound was 

studied at an initial concentration of 1 ppm, although the aqueous solubility is reported to be 0.2 

ppm at 20 C. Up to 60% of the applied was not characterized (i.e., isolated and identified) and 

degradates were not definitively identified. Additionally, the artificial light source used in the study 

was not fully described.

The 1986 adsorption/desorption batch equilibrium study was not conducted using 0.01 N CaCl2

solution, but was done with other salts which could alter the adsorption of the pesticide on the 

sediments studied. Freundlich adsorption coefficients were not calculated. Instead, adsorption was 

determined in five sediments each reportedly studied at four different pH levels.

A 1984 aerobic aquatic metabolism study (Acc#141274) is not valid, as material balances were 

variable; recoveries ranged from 73.2 to 173.1% of the applied in the nonsterile system and from 

120.4 to 188.3% of the applied in the sterile system. Additionally, not all major degradates were 

identified, and unextracted residues were high, at up to 86% of the applied in some samples, 

suggesting that the single extraction solution (acetone:water, 3:1 v:v) used to extract the sediment 

samples was not sufficient. Similar deficiencies were found in the other 1984 aerobic and anaerobic

aquatic metabolism studies (Acc# 253754).
17

Table 4. Submitted Environmental Fate Data for Rotenone.

OCSPP

Guideline Data Requirement

Submitted 

Studies

(Accession # 

or MRID)

Study 

Classification

Are data needed to 

conduct risk 

assessment?

Justification and 

Assumptions EPA will 

Make in Absence of 

Data

835.2120 Hydrolysis 141409 Not Acceptable Yes

In the absence of data, 

rotenone will be 

assumed to be stable.

835.2370 Aqueous Photolysis

254730

141410 Not Acceptable Yes

In the absence of data, 

rotenone will be 

assumed to be stable.

835.2410 Soil Photolysis

No data. Not 

required for 

aquatic use.  ̄ No  ̄

835.4100 Aerobic Soil 

Metabolism No data.  ̄ No

In the absence of data, 

rotenone will be 

assumed to be stable. 

835.4200 Anaerobic Soil 

Metabolism

No data. Not 

required for 

aquatic use.

 ̄ No  ̄

835.4300 Aerobic Aquatic 

Metabolism

141274

253454 Not Acceptable Yes

In the absence of data, 

rotenone will be 

assumed to be stable.

835.4400 Anaerobic Aquatic 

Metabolism

141273

(253754) Not Acceptable Yes

In the absence of data, 

rotenone will be 

assumed to be stable.

835.1230

Adsorption/ 

Desorption (batch 

equilibrium)

00157850 Not Acceptable Yes  ̄

835.6100 Aquatic Field 

Dissipation

Acc. # 

00157851 

(TRID 

470152-010)

MRID 

42217701

Supplemental No  ̄

850.1730 Bioaccumulation in 

Fish

Acc. #146183

Acc. #143252 Supplemental No  ̄

b. Ecological Effects
18

Table 5 identifies ecological effects studies with rotenone, by MRID (or Accession number), for 

each guideline requirement, as well as study classifications and whether or not additional data are 

needed in order to support risk assessment. Rationale for any additional data requested is presented 

below the tables. No data are available for the chronic effects of rotenone on birds; given that the 

compound is applied aerially and that there may be circumstances where the compound is not 

deactivated in lentic environments, the likelihood of chronic exposure to birds is uncertain; 

therefore, avian reproduction data are recommended. Data are also not available for terrestrial 

plants; however, given that rotenone is a flavonoid derived from plants and given that the 

compound is no longer applied to terrestrial environments, the likelihood of adverse effects on 

terrestrial plants from the use of rotenone as a piscicide is considered low.

No data are available for estuarine/marine organism; however, the compound is prohibited from 

use in estuarine/marine environment. The SOP also stipulates that when applied to freshwater 

environments processes (e.g., deactivation) should be in place to ensure that the compound does 

not move outside of the targeted treatment area; , therefore, the likelihood of exposure is considered 

low. While toxicity data are available for freshwater fish and invertebrates for both the technical 

grade active ingredient and multiple formulations (USEPA 2007), no data are available for the most 

recent formulation of rotenone (i.e., CFT LegumineTM 3.3%) which was developed to address 

uncertainties regarding the solvents (e.g., toluene) associated with older formulations. Therefore, 

EFED recommends acute toxicity data for the formulated product using rainbow trout (O. mykiss) 

(OSCPP Guideline 850.1075) and using the waterflea (D. magna) (OCSPP 850.1010). Since no 

data are available on the extent to which rotenone may partition to benthic sediments and given the 

estimated octanol-water partition coefficient (Kow≈4.10) and the potentially high organic carbon 

partition coefficient (Koc), EFED recommends the 10-day sediment toxicity test with both the 

midge (Chironomus riparius) and the amphipod (Hyalella azteca) (OCSPP Guideline 850.1735). 

Pending the outcome of the 10-day sediment toxicity test, a whole-sediment chronic toxicity test 

with a freshwater benthic invertebrate may be recommended as well; therefore, the whole sediment 

chronic toxicity test should be held in reserve.

Although rotenone is not applied to estuarine/marine environments, EFED is recommending acute 

toxicity data on molluscs (OCSPP Guideline 850.1025); however, rather than using the Eastern 

oyster (Crassostrea virginica), the registrant should consider substituting a freshwater mussel (e.g., 

Corbicula spp.).

In 2014, EPA in collaboration with Health Canada’s Pest Management Regulatory Agency and the 

California Department of Pesticide Regulation issued a harmonized guidance document on 

assessing the risk of pesticides to bees (USEPA 201438). In the guidance, multiple laboratory-based 

acute and chronic toxicity tests with individual honeybee adults and larvae were identified in 

addition to semi- and full-field tests with honey bee colonies to inform the risk assessment process. 

The tiered data requirements for honeybees identified in Table 5 reflect the full suite of studies 

identified in the guidance. Whether such tests are required though depends on the likelihood that 

bees will be exposed. Since rotenone is added directly to water, the likelihood of the material being 

applied to plants where bees may be foraging is considered low; therefore, these studies are not 

being required at this time.

 38 USEPA 2014. Guidance for Assessing Pesticide Risks to Bees. Office of Pesticide Programs, USEPA; Health Canada Pest 

Management Regulatory Agency, Ottawa, ON, Canada; California Department of Pesticide Regulation, Sacramento, CA. 

http://www2.epa.gov/sites/production/files/2014-06/documents/pollinator_risk_assessment_guidance_06_19_14.pdf
19

Table 5. Submitted Ecological Effects Data for Rotenone.

Guideline Description—Test 

Substance

MRID or 

Acc. No.

Study 

Classification

Are data 

Needed for 

Risk 

Assessment?

Comments

(Justification and Assumptions 

EPA will Make in Absence of 

Data)

Avian and Mammalian Testing

71-1

(850.2100)

Avian acute oral 

toxicity, waterfowl

Mallard duck

(Anas 

platyrhynchos)

TEP

Acc. No.

143250 Supplemental No Guideline specifies that acute 

oral toxicity test should be 

conducted using TGAI; however, 

given that the compound is 

applied directly to water and the 

likelihood of exposure is 

considered low; the currently 

available data on TEP are 

sufficient.

71-1

(850.2100)

Avian acute oral 

toxicity, upland 

game bird species

Ring-necked 

pheasant

(Phasianus 

colchicus)

TEP

Acc. No. 

143250

Supplemental No

850.2100

Avian acute oral 

toxicity, passerine 

species

TEP

No data No

Based on available data, rotenone 

is slightly toxic to practically 

non-toxic to birds on both an 

acute oral and subacute dietary 

exposure basis. Compound 

applied directly to water; 

therefore, the likelihood of 

exposure to passerine species 

considered low. Additional 

testing of birds is not needed.

850.2200

Avian dietary 

toxicity, waterfowl 

species

Mallard duck

(Anas platyrhynchos

TEP

Acc. No. 

248788 

(Hill et al., 

1975)

Supplemental No

Guideline specifies that acute 

oral toxicity test should be 

conducted using TGAI; however, 

given that the compound is 

applied directly to water and the 

likelihood of exposure is 

considered low; the currently 

available data on TEP are 

sufficient.

Avian dietary 

toxicity, upland 

game

Japanese quail

(Coturnix japonica)

bird

Ring-necked 

pheasant

(Phasianus 

colchicus)

TEP

Acc. No. 

248788 

(Hill et al., 

1975)

Supplemental No
20

Guideline Description—Test 

Substance

MRID or 

Acc. No.

Study 

Classification

Are data 

Needed for 

Risk 

Assessment?

Comments

(Justification and Assumptions 

EPA will Make in Absence of 

Data)

Avian dietary 

toxicity, upland 

game bird

TEP

Acc. No. 

248788

(Hill et al., 

1975)

Supplemental No

71-4

(850.2300)

Avian reproduction, 

waterfowl species—

TGAI

No data Yes

Based on available data, rotenone 

is slightly toxic to practically 

non-toxic to birds on both an 

acute oral and subacute dietary 

exposure basis. Rotenone is

added directly to water and may 

be deactivated; however, spray 

drift may be associated with the 

aerial application. Likelihood of 

chronic exposure is uncertain. In 

the absence of data, chronic risk 

to birds will be presumed.

71-4

(850.2300)

Avian reproduction, 

upland game bird 

species—TGAI

No data Yes

Based on available data, rotenone 

is slightly toxic to practically 

non-toxic to birds on both an 

acute oral and subacute dietary 

exposure basis. Rotenone is 

added directly to water and may 

be deactivated; however, spray 

drift maybe associated with the 

aerial application. Likelihood of 

chronic exposure is uncertain. In 

the absence of data, chronic risk 

to birds will be presumed.

850.2400 Wild mammal 

toxicity—TGAI No data No

Chronic toxicity data are already 

available and are considered 

sufficient for risk assessment.

850.2500 Simulated or actual 

field testing—TEP No Terrestrial uses eliminated.

Aquatic Animal Testing

72-2

(850.1010)

Freshwater 

invertebrate, acute 

toxicity

Waterflea

(Daphnia magna)

TGAI

400633-03 Supplemental No
21

Guideline Description—Test 

Substance

MRID or 

Acc. No.

Study 

Classification

Are data 

Needed for 

Risk 

Assessment?

Comments

(Justification and Assumptions 

EPA will Make in Absence of 

Data)

72-2

(850.1010)

Freshwater 

invertebrate, acute 

toxicity

Waterflea

(Daphnia magna)

TEP

See 

comment 

box to right

Yes

Although multiple test are 

available on formulated products, 

rotenone has recently been 

reformulated to reduce the 

reliance on organic solvents. 

Data are needed to evaluate the 

relative toxicity of the new 

formulation to freshwater 

invertebrates.

72-3

(850.1025)

Estuarine/Marine 

Mollusc acute 

toxicity—TGAI

No data Yes

Compound cannot be applied in 

estuarine/marine environments; 

however, freshwater mussels may 

be exposed to rotenone. Consider 

testing Corbicula sp. in lieu of 

Eastern oyster.

72-3

(850.1035)

Estuarine/Marine 

crustacean acute 

toxicity

TGAI

No data No Compound cannot be applied in 

estuarine/marine environments.

850.1075

Freshwater fish, 

acute toxicity, warm 

water species

Bluegill sunfish 

(Lepomis 

macrochirus) 

TGAI

439751-01

443829-02 Acceptable No

850.1075

Freshwater fish, 

acute toxicity, cold 

water species

Rainbow trout 

(Oncorhynchus 

mykiss)

TGAI

439751-02

443829-01

400633-01

Acceptable No

850.1075

Freshwater fish, 

acute toxicity, cold 

water species

Rainbow trout 

(Oncorhynchus 

mykiss)

TEP

See comment box to right. Yes

Although multiple test are 

available on formulated products, 

rotenone has recently been 

reformulated to reduce the 

reliance on organic solvents. 

Data are needed to evaluate the

relative toxicity of the new 

formulation to freshwater fish.

850.1075

Estuarine/Marine 

fish acute toxicity—

TGAI

No data No Compound cannot be applied in 

estuarine/marine environments.
22

Guideline Description—Test 

Substance

MRID or 

Acc. No.

Study 

Classification

Are data 

Needed for 

Risk 

Assessment?

Comments

(Justification and Assumptions 

EPA will Make in Absence of 

Data)

72-4

(850.1300)

Freshwater 

invertebrate,

reproduction test

Waterflea 

(Daphnia magna)

TGAI

400633-03 Supplemental No

850.1350

Estuarine/marine 

invertebrate,

reproduction test—

TGAI

No data No Compound cannot be applied in 

estuarine/marine environments.

850.1400

Freshwater fish, 

early life stage test

Rainbow trout

(Oncorhynchus 

mykiss)

TGAI

400633-02 Supplemental No

850.1400

Saltwater fish, early 

life stage test—

TGAI

No data No Compound cannot be applied in 

estuarine/marine environments.

850.1500

Freshwater fish life 

cycle test No data No

Estuarine/marine 

fish life cycle test No data No Compound cannot be applied in estuarine/marine environments.

850.1950

Simulated or actual 

field testing for 

aquatic organisms

Open 

literature Supplemental No

Lentsch et al. 200139 describes 

actual application of rotenone; 

Finlayson et al. 200140

Sediment Testing

850.1735

Whole sediment 10-

d freshwater 

invertebrate—TGAI

Midge (Chironomus 

riparius) and 

amphipod (Hyalella 

azteca)

No data Yes

Given the high Kow (estimated 

4.10) and uncertainty regarding 

the extent to which it partitions to 

sediment (i.e., unknown Kd and 

Koc) 

850.1740

Whole sediment 10-

d estuarine/marine 

invertebrate—TGAI

No data No Compound cannot be applied in 

estuarine/marine environments.

 39 Lentsch, L. D., C. W. Thompson, and R. L. Spateholts. 2001. Overview of Large-scale Chemical Treatment Success Story: Strawberry 

Valley, Utah. Pates 63 - to in R. L. Cailteux, L. DeMong, B. J. Finlayson, W. Horton, W. McClay, R. A. Schnick, and C. Thompson, 

editors. Rotenone in fisheries: are the rewards worth the risks? American Fisheries Society, Trends in Fisheries Science and 

Management 1, Bethesda, Maryland.

40 Finalyson, B. J., S. Siepmann, and J. Trumbo. 2001. Chemical Residues in Surface and Ground Waters Following Rotenone 

Application to California Lakes and Streams. Pages 37 to 53 in: R. L. Cailteux, L. DeMong, B. J. Finlayson, W. Horton, W. McClay, 

R. A. Schnick, and C. Thompson (editors), Rotenone in Fisheries: Are the Rewards Worth the Risks? American Fisheries Society, 

Trends in Fisheries Science and Management 1, Bethesda, MD.
23

Guideline Description—Test 

Substance

MRID or 

Acc. No.

Study 

Classification

Are data 

Needed for 

Risk 

Assessment?

Comments

(Justification and Assumptions 

EPA will Make in Absence of 

Data)

Nonguideline

Whole sediment 

chronic freshwater 

and/or marine 

invertebrate—TGAI

No data Yes

Whole sediment chronic study 

with freshwater invertebrates 

held in reserve pending outcome 

of 10-day sediment toxicity test 

(850.1735)

Insect Pollinator Testing

850.3020

Honeybee acute 

contact toxicity

(Apis mellifera)

TGAI

Acc. No.

05001991 

(Stevenson 

JH, 1978)

Acc. No. 

00036935 

(Atkins EL

et al., 1975)

Acceptable No

Nonguideline

(OECD 213)

Honeybee acute oral 

toxicity study—

TGAI 

No data No

Rotenone is added directly to 

water; likelihood of exposure is 

considered low.

Nonguideline

(OECD 237)

Honeybee Larval 

Acute Toxicity 

Test—TGAI

No data No

Rotenone is added directly to 

water; likelihood of exposure is 

considered low.

Nonguideline

Honeybee 10-day 

adult chronic 

toxicity

No data No

Rotenone is added directly to 

water; likelihood of chronic 

Non exposure is considered low. -

guideline

Honeybee 21-day 

larval chronic 

toxicity

No data No

850.3030

Honeybee toxicity of 

residues on 

foliage—TEP

No data No Based on acute contact toxicity 

data, study is not triggered.

850.3040 Field testing for 

pollinators No data No

Based on acute toxicity data and 

the absence of adverse effect 

data, study not required at this 

time.

Terrestrial Plant Testing

850.4100 Seedling emergence 

(10 species)—TEP 

No data

No Compound is applied directly to 

water and is a natural constituent 

of plants. 850.4150 Vegetative vigor (10 

species)—TEP No

Aquatic Plant Testing

850.4400

Aquatic plant 

growth, vascular 

plant 

—TGAI

No data No Compound is a natural 

constituent of plants.
24

Guideline Description—Test 

Substance

MRID or 

Acc. No.

Study 

Classification

Are data 

Needed for 

Risk 

Assessment?

Comments

(Justification and Assumptions 

EPA will Make in Absence of 

Data)

850.4500*

Aquatic Plant, 

freshwater green 

alga species—TGAI 

No data No

Compound is a natural 

constituent of plants.

Aquatic Plant, 

freshwater diatom

—TGAI 

No data No

Aquatic Plant, 

marine diatom

—TGAI

No data No

850.4550†

Aquatic Plant, 

cyanobacterium—

TGAI 

No data No

TGAI = Technical Grade Active Ingredient

TEP = technical end product

* Algal toxicity tests are now under guideline OCSPP 850.4500 (formerly 850.5400)

† Cyanobacterium tests are now under guideline 850.4550 (formerly 850.5400)
25

APPENDIX A. Submitted Guideline Studies

Environmental Fate

161-1 Hydrolysis

MRID Citation Reference

45950 Derse, P.H.; Strong, F.M. (1963) Toxicity of Antimycin to fish. Nature 200(4906): 

600-601. (Also~In~unpublished submission received Jun 30, 1965 under 8991-5; 

submitted by Ayerst Laboratories, Div. of American Home Products Corp., New 

York, N.Y.; CDL:100528-D) 

141409 2038363 Thomas, R. (1983) Hydrolysis of [6-[Carbon 14]]-Rotenone: Final Report: 

Borriston Project No. 0301A. Unpublished study prepared by Borriston 

Laboratories, Inc. 11 p. 

141731 2022496 Spare, W. (1984) Final Report: Hydrolysis of [Radiolabeled] Rotenone: Project 

No. 82-E-076-AM. Unpublished study prepared by Biospherics, Inc. 19 p

46688301 Alva, S. (2005) Giochem Cube Powder: Product Identity, Composition and 

Analysis. Unpublished study prepared by Universidad Nacional Agraria La 

Molina. 33 p

161-2 Photodegradation-water

MRID Citation Reference

141410 2022497 

2038364

Spare, W. (1982) Aqueous Photodegradation of [Carbon 14]-Rotenone: Final 

Report: Project No. 82-E-076-P. Unpublished study prepared by Biospherics 

Incorporated. 18 p. 

162-1 Aerobic soil metabolism

MRID Citation Reference

141411 Felkner, I. (1983) Effects of Microbes on Rotenone: Final Report: Borriston Project 

No. 0301B. Unpublished study prepared by Bor- riston Laboratories, Inc. 22 p. 

46688301 Alva, S. (2005) Giochem Cube Powder: Product Identity, Composition and Analysis. 

Unpublished study prepared by Universidad Nacional Agraria La Molina. 33 p.

162-2 Anaerobic soil metabolism
26

MRID Citation Reference

141411 Felkner, I. (1983) Effects of Microbes on Rotenone: Final Report: Borriston Project 

No. 0301B. Unpublished study prepared by Borriston Laboratories, Inc. 22 p. 

162-3 Anaerobic aquatic metabolism

MRID Citation Reference

254731 2022602 Felkner, I. (1983) Effects of Microbes on Rotenone: Final Report: Borriston 

Project No. 0301B. Unpublished study prepared by Borriston Laboratories, Inc. 22 

p.

141273 2022496 

2038365

Spare, W. (1984) Final Report: Anaerobic Aquatic Metabolism of [Radiolabeled] 

Rotenone: Project No. 82-E-076-AM. Unpublished study prepared by 

Biospherics, Inc. 19 p. 

162-4 Aerobic aquatic metabolism

MRID Citation Reference

254731 2022602 Felkner, I. (1983) Effects of Microbes on Rotenone: Final Report: Borriston 

Project No. 0301B. Unpublished study prepared by Borriston Laboratories, Inc. 22 

p.

141273 2022496 Spare, W. (1984) Final Report: Aerobic Aquatic Metabolism of [Radiolabeled] 

Rotenone: Project No. 82-E-076-AM. Unpublished study prepared by 

Biospherics, Inc. 19 p.

141274 2038366 Spare, W. (1984) Final Report: Aerobic Aquatic Metabolism of [Radiolabeled] 

Rotenone: Project No. 82-E-076-M. Unpublished study prepared by Biospherics, 

Inc. 21 p. 

163-1 Leach/adsorption/desorption

MRID Citation Reference

113292 2022598 

Ref

International Minerals & Chemical Corp. (1950) ?Dilan: Residues in Soil, Apples 

and Other Crops|. (Compilation; unpublished study received on unknown date 

under unknown admin. no.; CDL:120375-A) 

157850 2038366 

2038368

Dawson, V. (1986) Adsorption-desorption of [6a-[Carbon-14]]-Rotenone by 

Bottom Sediments: Final Report: ROT-84-988.02. Unpublished study prepared by 

National Fishery Research Laboratory, U.S. Fish and Wildlife Service. 144 p. 

46688301 Alva, S. (2005) Giochem Cube Powder: Product Identity, Composition and 

Analysis. Unpublished study prepared by Universidad Nacional Agraria La 

Molina. 33 p.
27

163-2 Volatility - lab

MRID Citation Reference

46688301 Alva, S. (2005) Giochem Cube Powder: Product Identity, Composition and 

Analysis. Unpublished study prepared by Universidad Nacional Agraria La 

Molina. 33 p.

163-3 Volatility - field

MRID Citation Reference

46688301 Alva, S. (2005) Giochem Cube Powder: Product Identity, Composition and 

Analysis. Unpublished study prepared by Universidad Nacional Agraria La 

Molina. 33 p.

164-2 Aquatic field dissipation

MRID Citation Reference

157850 2038367 

2038368

Dawson, V. (1986) Adsorption-desorption of [6a-[Carbon-14]]-Roten- one by 

Bottom Sediments: Final Report: ROT-84-988.02. Unpublished study prepared by 

National Fishery Research Laboratory, U.S. Fish and Wildlife Service. 144 p. 

157851 2038367 Gilderhus, P.; Dawson, V.; Allen, J. (1986) Persistence of Rotenone in Aquatic 

Environments at Different Temperatures: Final Report: Report No. ROT-83-

988.02. Unpublished study prepared by National by National Fishery Research 

Laboratory, U.S. Fish and Wild- life Service. 99 p. 

40307401 Fugami, J.; Yamamoto, I.; Casida, J. (1966) Metabolism of rotenone in vitro by 

tissue homogenates from mammals and insects. Scien- ce 155:713-716. 

40307402 Fugami, J.; Shishido, T.; Fukunaga, K.; et al. (1969) Oxidative metabolism of 

rotenone in mammals, fish, and insects and its relation to selective toxicity. 

Journal of Agricultural and Food Chemistry 17:1217-1226. 

40307406 2022596 

2022602 

2033061 

Ref

Dawson, V. (1986) Analyses of Pretreatment Samples: Addendum to Study, 

Persistence of Rotenone in Aquatic Environments at Different Temperatures: 

Project/Study No. ROT 83-988.02. Unpublished study prepared by U.S. Fish and 

Wildlife Service. 20 p. 

40307407 2022578 Dawson, V.; Allen, J. (1987) Liquid Chromatographic Determination of Rotenone 

in Fish, Crayfish, Clams, and Sediments: Laboratory Project ID: Project/Study 

No. ROT 83-988.02. Unpublished study prepared by U.S. Fish and Wildlife 

Service. 16 p. 

42217701 2050430 Finlayson, B.; Harrington, J. (1991) Chemical Residues in Surface and Ground 

Waters Following Rotenone Application to California Lakes and Streams. 

Unpublished study prepared by California Dept. of Fish and Game. 39 p. 
28

165-4 Bioaccumulation in fish

MRID Citation Reference

143252 2038367 Gingerich, W. (1984) Bioaccumulation Potential of Rotenone in Fish. 

Unpublished study prepared by US Fish and Wildlife Service. 5 p. 

146183 2038367 

2038370

Gingerich, W.; Rach, J. (1985) Accumulation and Elimination of [6A- Carbon14]-Rotenone by Bluegills: Final Report. Unpublished study prepared by U.S. 

Fish and Wildlife Service. 115 p.

40307401 Fugami, J.; Yamamoto, I.; Casida, J. (1966) Metabolism of rotenone in vitro by 

tissue homogenates from mammals and insects. Science 155:713-716. 

40307402 2022498 

2022602

Fugami, J.; Shishido, T.; Fukunaga, K.; et al. (1969) Oxidative metabolism of 

rotenone in mammals, fish, and insects and its relation to selective toxicity. 

Journal of Agricultural and Food Chemistry 17:1217-1226. 

40307403 Gingerich, W. (1987) Isolation, Purification, and Characterization of a Major 

Polar [Carbon 14]-Rotenoid Residue from Rainbow Trout Bile: Addendum: 

Project/Study No: ROT 82-977.03. Unpublished study prepared by U.S. Fish and 

Wildlife Service. 46 p. 

43494302 Gingrich, W. (1986) Distribution and accumulation of rotenone in tissues of 

warmwater fish. Transactions of the American Fisheries Society 115(1986):214-

219. 

43796401 Gilderhus, P.; Dawson, B.; Allen, J. (1984) Deposition and Persistence of 

Rotenone in Shallow Ponds During Cold and Warm Seasons: Supplement: (Raw 

Data): Lab Project Number: ROT-83-988.02. Unpublished study prepared by U.S. 

Fish and Wildlife Service. 1076 p. 

165-5 Bioaccum-aquatic non-target

MRID Citation Reference

146183 2038370 Gingerich, W.; Rach, J. (1985) Accumulation and Elimination of [6A- Carbon14]-Rotenone by Bluegills: Final Report. Unpublished study prepared by U.S. 

Fish and Wildlife Service. 115 p. 

157851 2038367 Gilderhus, P.; Dawson, V.; Allen, J. (1986) Persistence of Rotenone in Aquatic 

Environments at Different Temperatures: Final Report: Report No. ROT-83-

988.02. Unpublished study prepared by Nation- al by National Fishery Research 

Laboratory, U.S. Fish and Wild- life Service. 99 p. 

40307401 Fugami, J.; Yamamoto, I.; Casida, J. (1966) Metabolism of rotenone in vitro by 

tissue homogenates from mammals and insects. Science 155:713-716. 

40307402 2022498 Fugami, J.; Shishido, T.; Fukunaga, K.; et al. (1969) Oxidative metabolism of 

rotenone in mammals, fish, and insects and its relation to selective toxicity. 

Journal of Agricultural and Food Chemistry 17:1217-1226.
29

40307404 Yamamoto, I.; Unai, T.; Ohkawa, H.; et al. (1970) Stereochemical considerations 

in the formation and biological activity of the rotenone metabolites. Pesticides 

Biochemistry and Physiology 1:143-150. 

40307405 Gingerich, W.; Rach, J. (1984) Uptake, biotransformation, and elimination of 

rotenone by bluegills (lepomis macrochirus). Aquatic Toxicology 6:179-196. 

40307406 2022596 Dawson, V. (1986) Analyses of Pretreatment Samples: Addendum to Study, 

Persistence of Rotenone in Aquatic Environments at Different Temperatures: 

Project/Study No. ROT 83-988.02. Unpublished study prepared by U.S. Fish and 

Wildlife Service. 20 p. 

40307407 2022578 

Ref

Dawson, V.; Allen, J. (1987) Liquid Chromatographic Determination of Rotenone 

in Fish, Crayfish, Clams, and Sediments: Laboratory Project ID: Project/Study 

No. ROT 83-988.02. Unpublished study prepared by U.S. Fish and Wildlife 

Service. 16 p. 

40348709 2202602 Fugami, J.; Shishido, T.; Fukunaga, K.; et al. (1969) Oxidative metabolism of 

rotenone in mammals, fish, and insects and its relation to selective toxicity. 

Journal of Agricultural and Food Chemistry 17:1217-1226.

43494301 Gilderhus, P.; Dawson, B.; Allen, J. (1984) Deposition and persistence of 

rotenone in shallow ponds during cold and warm seasons. Journal of 

Investigations in Fish Control 95:1-7. 

43796401 Gilderhus, P.; Dawson, B.; Allen, J. (1984) Deposition and Persistence of 

Rotenone in Shallow Ponds During Cold and Warm Seasons: Supplement: (Raw 

Data): Lab Project Number: ROT-83-988.02. Unpublished study prepared by U.S

Fish and Wildlife Service. 1076 p

Non-guideline

MRID Citation Reference

46688301 Alva, S. (2005) Giochem Cube Powder: Product Identity, Composition and 

Analysis. Unpublished study prepared by Universidad Nacional Agraria La 

Molina. 33 p.

Ecological Effects

36935 Atkins, E. L., E. A. Greywood and R. L. Macdonald. 1975. Toxicity of Pesticides 

and Other Agricultural Chemicals to Honey Bees: Laboratory Studies. University 

of California, Department of Entomology, U.C. Cooperative Extension (Leaflet 

2287) 38p.

71003 Hudson, R. H., r. K. Tucker and M. A. Haegele. 1984. Handbook of Toxicity of 

Pesticides to Wildlife. USDI, Resource Publication 153. Washington DC, 90 p

141408 Kehoe, D, K. MacKenzie. 1983. Reproduction Study for Safety Evaluation of 

Rotenone Using Rats. Final Report. Study No 81077. Unpublished study 

prepared by Hazleton Raltech, Inc. 732p.
30

143250 Tucker, R. 1968. Rotenone: [Acute Oral Toxicity to Mallard Ducks and Ringnecked Pheasants]. Unpublished study prepared by U.S. Fish and Wildlife Service, 

Denver Wildlife Research Center. 8 p.

145496 Eiseman, J. and A. Thakur. 1984. General Metabolism Study for Safety 

Evaluation of Rotenone using Rats. Project No. 419-137. Unpublished Study 

Prepared by Hazleton Laboratories, America, Inc. 311 p.

248788 Hill, E. F., R. G. Heath, J. W. Spann and J. D. Williams. 1975. Lethal dietary 

toxicities of environmental pollutants to birds. U. S. Fish and Wildlife Service, 

Special Scientific Report Wildlife No. 191. Washington, D. C. 

400633-01 USFWS., 1986. Rotenone Freshwater Fish LC50 Rainbow Trout and Bluegill 

Sunfish. Lab ID Number 83-626.01B.

400633-02 Bills, T., J. Rach, and L. Marking. 1986. Rotenone--Fish Early Life Stage--

Rainbow Trout: Laboratory Project ID; TOX 83-626.01B. Unpublished study 

prepared by U.S. Fish and Wildlife Service. 15 p.

400633-03 Rach, J., T. Bills, and L. Marking. 1986. Rotenone--Acute LC50 Freshwater 

Invertebrate--Daphnia: Rotenone Aquatic Invertebrate Life Cycle-Daphnia: 

Laboratory Project ID; TOX 83-626/01A. Unpublished study prepared by U.S. 

Fish and Wildlife Service. 26 p.

439751-01 Sousa, J. 1996. Crystalline Rotenone--Acute Toxicity to Bluegill Sunfish (Lepomis

macrochirus) Under Flow-Through Conditions: Lab Project Number: 

10824.0695.6118.105: 95-10-6148: 051995/FIFRA/105. Unpublished study 

prepared by Springborn Laboratories, Inc. 70 p.

439751-02 Sousa, J. 1996. Crystalline Rotenone--Acute Toxicity to Rainbow Trout 

(Oncorhynchus mykiss) Under Flow-Through Conditions: Lab Project Number: 

10824.0625.6119.108: 95-10-6154: 051995/FIFRA/108. Unpublished study 

prepared by Springborn Laboratories, Inc. 70 p.

443829-01 Sousa, J. V., 1997. Rotenone Acute Toxicity to Rainbow Trout (Oncorhynchus 

mykiss) under Flow-through Conditions. Lab Report No. 97-9-7064. 

Unpublished study prepared by Springborn Laboratories, Wareham, MA.

443829-02 Sousa, J. V., 1997. Rotenone Acute Toxicity to Bluegill Sunfish (Lepomis 

macrochirus) under Flow-through Conditions. Lab Report No. 97-8-7063. 

Unpublished study prepared by Springborn Laboratories, Wareham, MA.
31

5001991 Stevenson, J. H. 1978. The Acute Toxicity of Unformulated Pesticides to Worker 

Honey Bees (Apis mellifera L.). Plant Pathology 27(1): 38 - 40.
32

APPENDIX B. EPI Suite Output File for Rotenone

SMILES : COc5cc4OCC3Oc2c1CC(Oc1ccc2C(=O)C3c4cc5OC)C(C)=C

CHEM : rotenone

MOL FOR: C23 H22 O6

MOL WT : 394.43

------------------------------ EPI SUMMARY (v4.11) --------------------

------

Physical Property Inputs:

Log Kow (octanol-water): ------

Boiling Point (deg C) : ------

Melting Point (deg C) : ------

Vapor Pressure (mm Hg) : ------

Water Solubility (mg/L): ------

Henry LC (atm-m3/mole) : ------

Log Octanol-Water Partition Coef (SRC):

Log Kow (KOWWIN v1.68 estimate) = 4.31

Log Kow (Exper. database match) = 4.10

Exper. Ref: HANSCH,C ET AL. (1995)

Boiling Pt, Melting Pt, Vapor Pressure Estimations (MPBPVP v1.43):

Boiling Pt (deg C): 501.20 (Adapted Stein & Brown method)

Melting Pt (deg C): 213.16 (Mean or Weighted MP)

VP(mm Hg,25 deg C): 6.94E-010 (Modified Grain method)

VP (Pa, 25 deg C) : 9.25E-008 (Modified Grain method)

MP (exp database): 176 deg C

BP (exp database): 210-220 @ 0.5 mm Hg deg C

Subcooled liquid VP: 2.56E-008 mm Hg (25 deg C, Mod-Grain method)

: 3.41E-006 Pa (25 deg C, Mod-Grain method)

Water Solubility Estimate from Log Kow (WSKOW v1.42):

Water Solubility at 25 deg C (mg/L): 1.045

log Kow used: 4.10 (expkow database)

no-melting pt equation used

Water Sol (Exper. database match) = 0.2 mg/L (20 deg C)

Exper. Ref: AUGUSTIJN-BECKERS,PWM ET AL. (1994)

Water Sol Estimate from Fragments:

CH3

O

O

O

O

O O

CH3

H3C

H2C
33

Wat Sol (v1.01 est) = 0.013116 mg/L

ECOSAR Class Program (ECOSAR v1.11):

Class(es) found:

Vinyl/Allyl Ethers

Henrys Law Constant (25 deg C) [HENRYWIN v3.20]:

Bond Method : 1.12E-013 atm-m3/mole (1.13E-008 Pa-m3/mole)

Group Method: Incomplete

For Henry LC Comparison Purposes:

User-Entered Henry LC: not entered

Henrys LC [via VP/WSol estimate using User-Entered or Estimated 

values]:

HLC: 3.447E-010 atm-m3/mole (3.492E-005 Pa-m3/mole)

VP: 6.94E-010 mm Hg (source: MPBPVP)

WS: 1.04 mg/L (source: WSKOWWIN)

Log Octanol-Air Partition Coefficient (25 deg C) [KOAWIN v1.10]:

Log Kow used: 4.10 (exp database)

Log Kaw used: -11.339 (HenryWin est)

Log Koa (KOAWIN v1.10 estimate): 15.439

Log Koa (experimental database): None

Probability of Rapid Biodegradation (BIOWIN v4.10):

Biowin1 (Linear Model) : 1.3355

Biowin2 (Non-Linear Model) : 0.9999

Expert Survey Biodegradation Results:

Biowin3 (Ultimate Survey Model): 1.8648 (months )

Biowin4 (Primary Survey Model) : 3.5050 (days-weeks )

MITI Biodegradation Probability:

Biowin5 (MITI Linear Model) : 0.6583

Biowin6 (MITI Non-Linear Model): 0.3312

Anaerobic Biodegradation Probability:

Biowin7 (Anaerobic Linear Model): 0.3225

Ready Biodegradability Prediction: NO

Hydrocarbon Biodegradation (BioHCwin v1.01):

Structure incompatible with current estimation method!

Sorption to aerosols (25 Dec C)[AEROWIN v1.00]:

Vapor pressure (liquid/subcooled): 3.41E-006 Pa (2.56E-008 mm Hg)

Log Koa (Koawin est ): 15.439

Kp (particle/gas partition coef. (m3/ug)):

Mackay model : 0.879

Octanol/air (Koa) model: 675

Fraction sorbed to airborne particulates (phi):

Junge-Pankow model : 0.969

Mackay model : 0.986

Octanol/air (Koa) model: 1

Atmospheric Oxidation (25 deg C) [AopWin v1.92]:

Hydroxyl Radicals Reaction:

OVERALL OH Rate Constant = 328.8113 E-12 cm3/molecule-sec

Half-Life = 0.033 Days (12-hr day; 1.5E6 OH/cm3)

Half-Life = 23.421 Min

Ozone Reaction:

OVERALL Ozone Rate Constant = 1.200000 E-17 cm3/molecule-sec
34

Half-Life = 0.955 Days (at 7E11 mol/cm3)

Half-Life = 22.920 Hrs

Fraction sorbed to airborne particulates (phi):

0.978 (Junge-Pankow, Mackay avg)

1 (Koa method)

Note: the sorbed fraction may be resistant to atmospheric oxidation

Soil Adsorption Coefficient (KOCWIN v2.00):

Koc : 2.611E+005 L/kg (MCI method)

Log Koc: 5.417 (MCI method)

Koc : 3165 L/kg (Kow method)

Log Koc: 3.500 (Kow method)

Aqueous Base/Acid-Catalyzed Hydrolysis (25 deg C) [HYDROWIN v2.00]:

Rate constants can NOT be estimated for this structure!

Bioaccumulation Estimates (BCFBAF v3.01):

Log BCF from regression-based method = 1.787 (BCF = 61.25 L/kg wet-wt)

Log Biotransformation Half-life (HL) = -1.0793 days (HL = 0.0833 days)

Log BCF Arnot-Gobas method (upper trophic) = 2.006 (BCF = 101.5)

Log BAF Arnot-Gobas method (upper trophic) = 2.006 (BAF = 101.5)

log Kow used: 4.10 (expkow database)

Volatilization from Water:

Henry LC: 1.12E-013 atm-m3/mole (estimated by Bond SAR Method)

Half-Life from Model River: 1.038E+010 hours (4.326E+008 days)

Half-Life from Model Lake : 1.133E+011 hours (4.719E+009 days)

Removal In Wastewater Treatment:

Total removal: 34.83 percent

Total biodegradation: 0.36 percent

Total sludge adsorption: 34.47 percent

Total to Air: 0.00 percent

(using 10000 hr Bio P,A,S)

Level III Fugacity Model:

Mass Amount Half-Life Emissions

(percent) (hr) (kg/hr)

Air 5.87e-006 0.755 1000

Water 2.89 1.44e+003 1000

Soil 53.6 2.88e+003 1000

Sediment 43.5 1.3e+004 0

Persistence Time: 4.9e+003 hr
35

Appendix C. Screening Imbibition Program Output.

Table 1. Inputs

Parameter Value

Chemical name Rotenone

Solubility (in water at 25oC; mg/L) 0.2

Mammalian LD50 (mg/kg-bw) 286

Mammalian test species laboratory rat

Body weight (g) of "other" mammalian species

Mammalian NOAEL (mg/kg-bw) 0.5

Mammalian test species laboratory rat

Body weight (g) of "other" mammalian species

Avian LD50 (mg/kg-bw) 2200

Avian test species mallard duck

Body weight (g) of "other" avian species

Mineau scaling factor 1.15

Mallard NOAEC (mg/kg-diet) 0

Bobwhite quail NOAEC (mg/kg-diet) 0

NOAEC (mg/kg-diet) for other bird species 0

Body weight (g) of other avian species

NOAEC (mg/kg-diet) for 2nd other bird species

Body weight (g) of 2nd other avian species

Table 2. Mammalian Results

Parameter Acute Chronic

Upper bound exposure (mg/kg-bw) 0.0344 0.0344

Adjusted toxicity value (mg/kg-bw) 219.9799 0.3846

Ratio of exposure to toxicity 0.0002 0.0894

Conclusion*

Drinking water exposure 

alone is NOT a potential 

concern for mammals

Drinking water exposure 

alone is NOT a potential 

concern for mammals

Table 3. Avian Results

Parameter Acute Chronic

Upper bound exposure (mg/kg-bw) 0.1620 0.1620

Adjusted toxicity value (mg/kg-bw) 1142.2959 0.0000

Ratio of exposure to acute toxicity 0.0001 0.0000

Conclusion*

Drinking water exposure 

alone is NOT a potential 

concern for birds

Due to insufficient data, 

risk cannot be precluded

*Conclusion is for drinking water exposure alone. This does not combine all routes of exposure. Therefore, 

when aggregated with other routes (i.e., diet, inhalation, dermal), pesticide exposure through drinking water 

may contribute to a total exposure that has potential for effects to non-target animals.