Docket Number EPA-HQ-OPP-2015-0256 

www.regulations.gov 

Busan 77 Final Work Plan 

Registration Review: Initial Docket 

Case Number 3034 

March 2016 

Approved by: ~ Steve Knizner 

Director 

Antimicrobials Division 

Date: 3 /l'i!J 6 

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Busan 77 Registration Review Team

Human Health & Environmental Effects

Nathan Mottl

Julie VanAlstine

Michelle Centra

David Bays

Jim Breithaupt

Siroos Mostaghimi

Risk Management

Stephen Savage

Rose Kyprianou

Jacqueline Hardy

Stacey Grigsby
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TERMS, ABBREVIATIONS AND SYMBOLS

A.I. or a.i. active ingredient

ASRI activated sludge respiration inhibition

atm-m3

/mole atmospheric pressure-cubic meter per mole

°C degrees Celsius

CAS Chemical Abstracts Service 

CFR Code of Federal Regulations 

CHO Chinese hamster ovary

CMA Chemical Manufacturers Association

CO2 carbon dioxide

COC concentration-of-concern

cPAD chronic population adjusted dose 

DCI data call-in

EC50 median (or 50 percent) effect concentration

ECOTOX ECOTOXicology

EDI estimated daily intake

EDSP Endocrine Disruptor Screening Program 

E-FAST Exposure and Fate Assessment Screening Tool

EPA Environmental Protection Agency

FDA Food and Drug Administration 

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FQPA Food Quality Protection Act 

FWP Final Work Plan

g/mol grams per mole

GLN guideline number

IDS Incident Data System 

Koc organic carbon normalized soil-water partition coefficient

Kow octanol-water partition coefficient

LC50 median (or 50 percent) lethal concentration

LD50 median (or 50 percent) lethal dose

LOAEC lowest-observed-adverse-effect-concentration 

LOEC lowest-observed-effect-concentration

LOAEL lowest-observed-adverse-effect-level

Log Kow logarithm of the octanol-water partition coefficient

μg microgram

ml/g milliliter per gram

mg/kg milligram per kilogram

mg/kg/day milligram per kilogram per day 

mg/L milligram per liter

mm Hg millimeter of mercury

MOE margin of exposure 

MRID Master Record Identification Number

MRL maximum residue limit 

N/A not applicable 

nm nanometers

NOAEC no-observed-adverse-effect-concentration

NOAEL no-observed-adverse-effect-level

OCSPP Office of Chemical Safety and Pollution Prevention
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OECD Organization for Economic Co-operation and Development

OPP Office of Pesticide Programs

PAD population adjusted dose

PDM Probabilistic Dilution Model

PC Code Pesticide Chemical Code

pH power of hydrogen or power of the concentration of the hydrogen ion

ppb parts per billion 

ppm parts per million

PWP Preliminary Work Plan

QSAR quantitative structure-activity relationship

RED Reregistration Eligibility Decision

SAR structure activity relationship

SF safety factor

TEP typical end-use product

TGAI technical grade active ingredient

TMDL total maximum daily loads

UF uncertainty factor

UV/VIS ultraviolet/visible light absorption

% w/w percent weight per weight.

WP wettable powder

WWTPs wastewater treatment plants
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Table of Contents

1 Introduction............................................................................................................................. 7

1.1 Statutory and Regulatory Authority ................................................................................. 7

1.2 Updates to the Work Plan................................................................................................. 7

1.3 Case Overview ................................................................................................................. 8

1.4 Chemical Identification and Properties............................................................................ 9

1.5 Use/Usage Description................................................................................................... 10

1.5.1 Registrations ........................................................................................................... 10

1.5.2 Summary of Registered Uses.................................................................................. 10

1.5.3 Usage Information .................................................................................................. 11

1.6 Regulatory History ......................................................................................................... 11

1.6.1 Tolerance Information ............................................................................................ 12

1.7 Incidents......................................................................................................................... 12

1.7.1 Human Health ......................................................................................................... 12

1.7.2 Ecological ............................................................................................................... 12

2 Anticipated Data Needs ........................................................................................................ 12

3 Human Health Risk Assessment........................................................................................... 16

3.1 Existing Toxicological Endpoints.................................................................................. 16

3.2 Dietary Exposure............................................................................................................ 18

3.2.1 Food ........................................................................................................................ 18

3.2.2 Drinking Water ....................................................................................................... 18

3.3 Occupational and Residential Exposures....................................................................... 18

3.3.1 Occupational Handler Exposure ............................................................................. 19

3.3.2 Residential Handler Exposures............................................................................... 19

3.3.3 Residential Post-Application Exposures................................................................. 20

3.4 Aggregate and Cumulative Exposure............................................................................. 20

3.4.1 Aggregate Exposures.............................................................................................. 20

3.4.2 Cumulative Exposures............................................................................................ 20

4 Environmental Risk Assessment........................................................................................... 21

4.1 Environmental Fate Assessment .................................................................................... 21

4.1.1 Water Quality.......................................................................................................... 22

4.2 Conceptual Models for Environmental Exposure Pathways.......................................... 22

4.3 Ecological Effects Assessment....................................................................................... 23

4.3.1 Measures of Effect (Ecotoxicology Endpoints)...................................................... 23

4.4 Exposure Analysis Plan.................................................................................................. 24

4.4.1 Aquatic and Terrestrial Wildlife Exposure Estimates ............................................ 24

4.4.2 Screening Level Down-the-Drain Analysis............................................................ 24

4.5 Effects Analysis Plan ..................................................................................................... 24

5 Endocrine Disruptor Screening Program (EDSP)................................................................. 25

6 Optional Label Changes........................................................................................................ 26

7 Next Steps............................................................................................................................. 26

8 References............................................................................................................................. 27

Appendix A Toxicology Profile............................................................................................... 28

Appendix B Environmental Fate ............................................................................................. 47

Appendix C Ecotoxicology Profile.......................................................................................... 50
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Appendix D Screening Level Down-the-Drain Analysis ........................................................ 61

Appendix E Product Chemistry............................................................................................... 62

Appendix F Comments Received Concerning the Preliminary Work Plan............................ 64

List of Tables

Table 1 – Summary of Anticipated Risk Assessments and Data Needs for Registration Review: 

Busan 77.......................................................................................................................................... 8

Table 2 – Anticipated Registration Review Schedule .................................................................... 9

Table 3 – Chemical Identification of Busan 77 .............................................................................. 9

Table 4 – Physical-Chemical and Environmental Fate Properties for Busan 77.......................... 10

Table 5 – Summary of Busan 77 Registered Uses........................................................................ 11

Table 6 – Studies Anticipated as Needed for the Registration Review of Busan 77.................... 13

Table 7 – Data Needs Required by Data Call-In GDCI-069183-30193....................................... 14

Table 8 – Existing Toxicological Endpoints................................................................................. 16

Table 9 – Occupational Handler Exposure Scenarios for Busan 77............................................. 19

Table 10 – Residential Handler Exposure Scenarios for Busan 77 .............................................. 19

Table 11 – Residential Post-Application Exposure Scenarios for Busan 77 ................................ 20

Table 12 – Ecological Effects Endpoints Selected for Assessing Risks from Busan 77.............. 23
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1 Introduction

This document is the United States Environmental Protection Agency’s (USEPA, EPA or “the 

Agency”) Final Work Plan (FWP) for poly[oxy-1,2-ethanediyl(dimethyliminio)-1,2-

ethanediyl(dimethyliminio)-1,2-ethanediylchloride (1:2)], herein referred to as Busan 77. The 

FWP document explains what EPA's Office of Pesticide Programs (OPP) knows about Busan 77, 

highlighting anticipated data and assessment needs, identifying the types of information that 

would be especially useful to the Agency in conducting the review, and providing an anticipated 

timeline for completing Busan 77’s review.

The registration review process was designed to include a public participation component to 

solicit input from interested stakeholders. The Agency intends, by sharing this information in the 

docket, to inform the public of what it knows about Busan 77 and what types of new data or 

other information would be helpful for the Agency to receive as it moves toward a decision on 

Busan 77.

1.1 Statutory and Regulatory Authority

The Food Quality Protection Act (FQPA) of 1996 mandated a registration review program. All 

pesticides distributed or sold in the United States generally must be registered by the USEPA 

based on scientific data showing that they will not cause unreasonable risks to human health or 

the environment when used as directed on product labeling. The registration review program is 

intended to make sure that, as the ability to assess risk evolves and as policies and practices 

change, all registered pesticides continue to meet the statutory standard of no unreasonable 

adverse effects to human health or the environment. Changes in science, public policy, and 

pesticide use practices will occur over time. Through the registration review program, the 

Agency periodically reevaluates pesticides to make sure that as change occurs, products in the 

marketplace can be used safely. Information on this program is provided at 

http://www2.epa.gov/pesticide-reevaluation.

The Agency is implementing the registration review program pursuant to Section 3(g) of the 

Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) and will review each registered 

pesticide every 15 years to determine whether it continues to meet the FIFRA standard for 

registration. The regulations governing registration review begin at 40 CFR 155.40. The Agency

will consider benefits information and data as required by FIFRA. The public phase of 

registration review begins when the initial docket is opened for each case. The docket is the 

Agency’s opportunity to state what it knows about the pesticide and what additional risk analyses 

and data or information it believes are needed to make a registration review decision.

1.2 Updates to the Work Plan

Since the publication of the Preliminary Work Plan (PWP), the Agency has made the following 

updates:

• Updated the timeline in Table 2, “Anticipated Registration Review Schedule”

• Revised timeframe for guideline study 870.3465 in Table 6
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• Updated Section 3.4.2 hyperlink for, “Cumulative Exposures”

• Updated Section 7, “Next Steps”

• Deleted the “Guidance for Commenters” 

• Added Appendix F, “Comments Received Concerning the Preliminary Work Plan”

The Agency received two submissions during the public comment period on the initial docket. 

See Appendix F for the Agency’s response to these comments.

No modifications were made to the anticipated data needs and there is no change to the 

registration review schedule of Busan 77. This document makes final the work plan for the 

Busan 77 registration review process.

1.3 Case Overview

The docket for Busan 77 (case 3034) has been established at http://www.regulations.gov in 

docket number EPA-HQ-2015-0256. Documents associated with this registration review can be 

viewed in this docket. Tables 1 and 2 below summarize the issues relevant to this registration 

review case and the anticipated registration review schedule.

Table 1 – Summary of Anticipated Risk Assessments and Data Needs for Registration 

Review: Busan 77

Risk Assessment 

Assessment 

Necessary to 

Support 

Registration 

Review

Date of Most 

Recent 

Assessment

Type of 

Assessment 

Required 

(New/Updated)

Data Anticipated as Needed

(See Table 6 for details)*

Dietary (food) No 07/27/07 None None

Dietary (drinking water) Yes 07/27/07 Updated See Table 6

Occupational Handler Yes 07/27/07 Updated Product Use (875.1700)

Occupational Postapplication No N/A None None

Residential Handler Yes 07/27/07 Updated Product Use (875.2700)

Residential Post 

Application (Incidental 

Oral and Dermal)

Yes 07/27/07 Updated See Table 6

Aggregate Yes N/A New None 

Cumulative No N/A None None

Tolerance Review No 07/27/07 None None

Ecological Yes N/A New

ASRI, sorption to soil and sludge, 

Aquatic invertebrate life-cycle study

Freshwater duckweed study

N/A = Not applicable

* Table 1 is a summary of the anticipated risk assessments and data needs for Registration Review. For data called in through the 

post-Reregistration Eligibility Decision (RED) Data Call-In (DCI), see Table 7.
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Table 2 – Anticipated Registration Review Schedule

Anticipated Activity Target Date* Completion Date

Phase 1: Opening the Docket 

Open Docket and 60-Day Comment Period for Preliminary Work Plan 2015-09 2015-10-02

Close Public Comment Period 2015-11 2015-12-01

Phase 2: Case Development 

Issue Final Work Plan 2016-03 2016-03

Issue Data Call-In (DCI) 2017-03

Receive Data to be Considered in Risk Assessment 2019-03

Open 30-Day Public Comment Period for Preliminary Risk Assessment(s) 2020-09

Close Public Comment Period 2020-10

Phase 3: Registration Review Decision and Implementation 

Open 60-Day Public Comment Period for Proposed Decision 2021-03

Close Public Comment Period 2021-05

Issue Final Decision 2021-09

Begin Post-Decision Follow-up 2021

Total (years) 6

*The anticipated schedule will be revised as necessary (e.g., need arising under the Endocrine Disruptor Screening Program with 

respect to the active ingredients in this case).

1.4 Chemical Identification and Properties

Table 3 presents the active ingredient to be assessed in case 3034: Busan 77 (PC Code 069183). 

Table 3 – Chemical Identification of Busan 77

Chemical Name Busan 77

Chemical Classification Polymeric quaternary ammonium

Chemical Name Poly[oxy-1,2-ethanediyl(dimethyliminio)-1,2-ethanediyl(dimethyliminio)-1,2-

ethanediyl dichloride (1:2)]

PC Code 069183

CAS Number 31512-74-0

Molecular Formula C10H25N2O (monomer without chlorines)

Molecular Weight 

(grams/mole)

189.32 (monomer)1

3,368 (polymer)2

 1 EPIWEB 4.11

2 Average, 7/10/91 Buckman letter 
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Molecular Structure

Smiles Code CCOCCN(C)(C)CCN(C)(C) [includes 2 chlorines as counter ions]

The Busan 77 product chemistry information relevant to risk assessment is summarized in Table 

4 and the details of the product chemistry information are discussed in Appendix E.

Table 4 – Physical-Chemical and Environmental Fate Properties for Busan 77

Guideline No./

Study Type Results Reference/Comments (MRID unless specified)

Water Solubility(mg/L) 10,000 Assumed based on miscibility

830.7050

UV visible sorption N/A 49176301

Compound has no double bonds to absorb UV light

830.7370

Dissociation constant Waived 47120304/42372401

830.7550

Partition coefficient Waived 47120304

830.7950

Vapor pressure Negligible 47120304Manufacturing Use Product is 60 % polymer and 40 % water

Henry’s Law Constant 8.5 x 10-12 Volatility from water not likely to occur given the high MW, 

solubility, and positive charges that will enhance sorption to solids. 

1.5 Use/Usage Description

1.5.1 Registrations

There are 98 EPA-registered products that contain Busan 77 as an active ingredient (a.i.). The 

products include one product that contains 5-chloro-2-methyl-3(2H)-isothiazolinone (CMIT) and 

2-methyl-3(2H)-isothiazolinone (MIT), and other products that contain other a.i.’s. The percent 

a.i. ranges from 1.7 percent to 60 percent and the formulations include: ready-to-use, flowable 

concentrates, dry flowables, soluble concentrates and granulars. There are 2 manufacturing use 

products (EPA Registration Number 1448-32 and 83451-24) that contain 60 percent Busan 77.

1.5.2 Summary of Registered Uses

Busan 77 is currently registered as an active ingredient in end use products as an algaecide, 

bacteriostat, fungicide, microbiocide/microbiostat and molluscicide. Products containing Busan 

77 are used in swimming pools, spas, whirlpools, hot tubs, metal working fluids, fire water 

protection systems, cooling water towers, petroleum secondary recovery systems, paper mill 

process water, air washer water systems, and ornamental ponds. Table 5 presents a summary of 

the registered uses of Busan 77 that will be assessed in this registration review. Busan 77 can be 

applied by the following application methods: liquid pour, liquid pump, solid pour, and solid 

place.
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Table 5 – Summary of Busan 77 Registered Uses

Use Application Method Application Rate

Industrial Processes and Water Systems

Air Washer Systems Liquid Pour 45 ppm Liquid Pump

Heat Exchanger Water Systems Liquid Pump 18 ppm

Cooling Water Liquid Pour 34 ppm Liquid Pump

Secondary Oil Recovery Liquid Pour 45 ppm Liquid Pump

Aquatic Areas

Aquatic Areas Liquid Pour 45 ppm Liquid Pump

Ornamental Ponds Liquid Pour 11 ppm Liquid Pump

Swimming Pools

Swimming Pools, Exterior Spas, 

Whirlpools and Hot Tubs

Liquid Pour

9 ppm Liquid Pump

Solid Pour

Solid Place

Material Preservatives

Metalworking Fluids Liquid Pour 600 ppm Liquid Pump

Pulp and Paper Liquid Pour 0.26 lb. a.i./ton

Liquid Pump (150 ppm)

* Only maximum application rates are captured.

1.5.3 Usage Information

EPA did not find any specific distribution of usage or market data in the Kline Report for Busan 

77 (Kline, 2004).

1.6 Regulatory History

The first product containing Busan 77 as an active ingredient was registered in the United States 

in 1971. The Agency completed a Reregistration Eligibility Decision (RED) for Busan 77 in 

2007. The post-RED Data Call-In (DCI) was issued in September 2011. Products containing 

Busan 77 were previously registered for use in textiles and the last product with this use was 

cancelled.
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1.6.1 Tolerance Information

EPA has not established tolerances for residues of Busan 77 in or on raw agricultural

commodities or processed food and feed products under the Federal Food, Drug, and Cosmetic 

Act (FFDCA) Section 408. There are also no tolerance exemptions for residues of Busan 77 

under FFDCA Section 408. Busan 77 has been cleared as an indirect food additive by the US 

Food and Drug Administration (US FDA) under FFDCA Section 409 as a component of paper 

and paperboard in contact with aqueous and fatty foods (21CFR§176.170).

1.7 Incidents

1.7.1 Human Health 

Thirty four incidents were reported in Incident Data System (IDS) for the RED from the use of 

Busan 77; although limited, incidents reported in OPP IDS support that exposure to Busan 77 may 

cause some irritation through dermal, inhalation, and/or ocular exposures. Most of the symptoms are 

considered minor. The most common symptoms reported for cases of dermal exposure were skin 

irritation/burning, rash, itching and skin discoloration/redness. The most common symptoms reported 

for cases of inhalation exposure were minor respiratory irritation/burning. In some cases, a headache 

is reported after inhalation exposure. The symptoms associated with ocular exposure include blurred 

vision, edema and irritation around eyes. Since the date of the RED (12/2007), there were two

additional human health incidents listed in the OPP IDS for the period 12/2007 to present (search 

conducted on 5/28/2015). These new reported incidents were minor.

1.7.2 Ecological

No ecological incidents have been reported in the Agency’s Ecological Incident Information 

System (EIIS) from 2005 until the present (search conducted on 7/16/2015).

2 Anticipated Data Needs

Several studies were required from the assessment of Busan 77 in the 2007 Reregistration 

Eligibility Decision (RED). In addition to the already-required RED DCI data, the following 

studies as listed in Table 6 are expected to be needed for the registration review of Busan 77. 

Table 7 outlines the data requirements required by the post-RED DCI GDCI-069183-30193

issued on September 29, 2011. The Agency anticipates reviewing any data received in response 

to the post-RED DCI as well as data anticipated to be requested for this registration review prior 

to conducting the registration review risk assessments for Busan 77. 
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Table 6 – Studies Anticipated as Needed for the Registration Review of Busan 77

GLN Study Name Test Substance

Time Frame 

(Measured in months 

from DCI Receipt)

Risk Assessment(s) Data 

Will Support

Use Site(s) Triggering 

Anticipated Data 

Requirement

Applicable 

Exposure 

Scenario

835.1110 Activated sludge sorption isotherm1, 2 TGAI 12 Ecological

Recirculating cooling 

towers, air washer systems, 

petroleum secondary 

recovery systems, and 

metalworking fluids

Ecological 

exposure

835.1230 Leaching-adsorption-desorption TGAI 12 Ecological Ecological 

exposure

850.3300 Activated sludge respiration inhibition2, 3, 4 TGAI 12 Ecological Ecological 

exposure

850.1300 Aquatic invertebrate life-cycle study TGAI 12 Ecological Ecological 

exposure

850.4400 Freshwater floating macrophyte duckweed 

study TGAI 12 Ecological Ecological 

exposure

870.3465 90-Day Inhalation Toxicity-rat TGAI 24 Human Health

Recirculating cooling 

towers, air washer systems, 

petroleum secondary 

recovery systems,

metalworking fluids, 

swimming pools

Occupational 

exposure

and

Residential 

exposure

870.5300

or

870.5375

In Vitro Mammalian Gene Mutation TGAI 12

Human Health 

(Weight of Evidence 

for Mutagenicity)

Occupational 

exposure

and

Residential 

exposure

870.6200

Acute Neurotoxicity Screening Battery-rat; 

90-Day Neurotoxicity Screening Batteryrat5, 6TGAI 24 Human Health

Occupational 

exposure

and

Residential 

exposure

870.7800 Immunotoxicity TGAI 12 Human Health

Occupational 

exposure

and

Residential 

exposure

875.1700 Product Use EP 12 Human Health Recirculating cooling 

towers, air washer systems, 

Occupational 

exposure
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GLN Study Name Test Substance

Time Frame 

(Measured in months 

from DCI Receipt)

Risk Assessment(s) Data 

Will Support

Use Site(s) Triggering 

Anticipated Data 

Requirement

Applicable 

Exposure 

Scenario

petroleum secondary 

recovery systems, and 

metalworking fluids

Swimming pools and spas Residential 

exposure

875.2700 Product Use EP 12 Human Health Swimming pools and spas Residential 

exposure

1 EPA has a published final guideline for this study: http://www.regulations.gov/#!documentDetail;D=EPA-HQ-OPPT-2009-0152-0003.

2 Anticipated to be required because registered uses lead to Down-the-Drain (DtD) exposure.

3 EPA published draft guidance under guideline 850.6800 and has since published final guidance for this study under guideline 850.3300: 

http://www.regulations.gov/#!documentDetail;D=EPA-HQ-OPPT-2009-0154-0021.

4 OECD Test Guideline 209 can also be used as guidance for this study, available online at http://www.oecd-ilibrary.org/content/book/9789264070080-en.

5 Acute (study due 12 months after receipt of DCI) and subchronic (study due 24 months after receipt of DCI) neurotoxicity studies are anticipated to be required.

6 A protocol should be submitted for approval prior to testing.

Table 7 – Data Needs Required by Data Call-In GDCI-069183-30193 

GLN Study Name Test Substance Risk Assessment(s) Data Will 

Support

Use Site(s) Triggering Anticipated 

Data Requirement Status

850.1300 Daphnid Chronic Toxicity Test TGAI Ecological

Recirculating cooling towers, air 

washer systems, petroleum 

secondary recovery systems,

metalworking fluids, swimming 

pools and textiles1

Unsatisfied

850.1740 Whole sediment: Acute Marine 

Invertebrates TGAI Ecological MRID 49300901-Underreview

850.1400 Fish early-life stage study TGAI Ecological MRID 49362401-

Satisfied

850.1735 Whole sediment, acute invertebrates 

(freshwater) TGAI Ecological MRID 48715801-Satisfied

850.4225 Seedling emergence, Tier II Rice TGAI Ecological Unsatisfied
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GLN Study Name Test Substance Risk Assessment(s) Data Will 

Support

Use Site(s) Triggering Anticipated 

Data Requirement Status

850.4400 Aquatic Plant Toxicity Test using Lemna 

spp., Tiers I & II TGAI Ecological MRID 49327001 –Under review

850.5400 Algal Toxicity, Tiers I & II TGAI Ecological MRID 49453501-

Partially satisfied1

875.1100 Dermal Exposure, Outdoor EP NA NA Waived

875.1200 Dermal Exposure, Indoor EP Occupational Handler

Residential Handler All Unsatisfied

875.1300 Inhalation Exposure, Outdoor EP NA NA Waived

875.1400 Inhalation Exposure, Indoor EP Occupational Handler

Residential Handler All Unsatisfied

875.1600 Application Exposure Monitoring Data 

Reporting NA NA NA Waived

875.2300 Indoor Surface Residue Dissipation2 EP Residential Post-application Textiles Unsatisfied

SS-AFM-1 Aquatic Field Monitoring, Once-Through 

Cooling Water Systems

Once-Through Cooling Water 

Systems Unsatisfied

Footnotes

NA = not applicable

1. Guideline recommends green algae and diatom study. 

2. Products containing Busan 77 were previously registered for use in textiles and the last product with this use was voluntarily cancelled (18-Jul-2006). The GDCI is still listed as unsatisfied 

but data will no longer be required for 875.2300. 
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3 Human Health Risk Assessment

The Agency anticipates the need to conduct a human health risk assessment for Busan 77. The 

Agency expects to require additional data for use in conducting the registration review.

3.1 Existing Toxicological Endpoints

EPA anticipates the need to revise the existing toxicological endpoints as part of this registration 

review. Table 8 presents the endpoints used in EPA’s most recent human health risk assessment 

in support of the 2007 RED. Although mammalian toxicological studies were not identified as 

data gaps in the RED, four toxicity studies are identified in the registration review as anticipated 

data needs based on currently registered uses. All available information including existing 

toxicology studies, valid scientific literature, and the studies that are expected to be required for 

registration review will be considered in establishing the endpoints to be used in the registration 

review risk assessment. Appendix A provides a detailed description of the toxicity studies.

Table 8 – Existing Toxicological Endpoints

Exposure 

Scenario

Dose Used in Risk 

Assessment 

(mg/kg/day)

Target Margin of Exposure 

(MOE), Uncertainty Factor (UF), 

FQPA Safety Factor (SF)

Study and Toxicological Effects

Acute Dietary

(females 13+)

A single-dose effect in developing animals was not identified in the toxicity data base for Busan 

77 and therefore, this risk assessment is not required.

Acute Dietary

(gen pop)

A single-dose effect in adult animals was not identified in the toxicity data base for Busan 77 

and therefore, this risk assessment is not required.

Chronic Dietary

(gen pop)

NOAEL = 100 

mg/kg/day

UFA = 10

UFH = 10

Total UF = 100

FQPA SF = 1

Chronic RfD = NOAEL ÷ Total UF 

= 100 ÷ 100 = 1.0 mg/kg/day

Chronic cPAD = Chronic RfD ÷ 

FQPA = 1 ÷ 1 = 1 mg/kg/day

Chronic Toxicity/Carcinogenicity -

Rats

(MRID 41809101/41561301)

LOAEL = 300 mg/kg/day based upon 

body weight gain (M), reduced total 

protein (M), reduced inorganic 

phosphorus (M&F), increased urine

pH (F) and a dose related-increase in 

bloody crusts.

Incidental Oral 

Short-Term

(1 – 30 Days)

NOAEL = 500 

mg/kg/day

UFA = 10

UFH = 10

Target MOE = 100

Prenatal Developmental Toxicity -

Rats (MRID 41423001/93062018)

LOAEL = 700 mg/kg/day based upon 

increased mortality and morbidity 

(convulsions, lethargy and 

pathological changes in lung, brain, 

stomach and intestines).

Incidental Oral 

Intermediate-Term

(1 – 6 Months)

NOAEL = 221 

mg/kg/day

UFA = 10

UFH = 10

Target MOE = 100

Subchronic Oral Toxicity – Rats

(MRID 40025001/93662014)

LOAEL = 752 mg/kg/day based upon 

mineralization of renal tubules with 

decreases in body weights, absolute 

organ weights (heart, liver, kidney and 

gonads), equivocal decrease in RBC 
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Exposure 

Scenario

Dose Used in Risk 

Assessment 

(mg/kg/day)

Target Margin of Exposure 

(MOE), Uncertainty Factor (UF), 

FQPA Safety Factor (SF)

Study and Toxicological Effects

counts, elevated leukocyte counts, 

non-suppurative inflammation in the 

chorid plexus of the brain and 

increased animal death observed at the 

higher doses of 2554 and 3685 

mg/kg/day.

Dermal

Short-Term

(1 – 30 Days)

NOAEL = 10 

mg/kg/day

(125 μg/cm2; a

TGAI based dermal 

endpoint = 10 

mg/kg x 0.2 kg rat x 

1000 μg/mg) ÷ 16 

cm2)

UFA = 10

UFH = 10

Target MOE = 100

Subchronic Dermal Toxicity – Rats

(MRID 40170601)

LOAEL = 100 mg/kg/day based upon 

treatment-related dermatological 

changes consistent with chronic 

irritation/inflammation. The skin 

lesions consisted of one or more 

combinations of ulceration of the 

epidermis, chronic inflammation of 

the dermis, acanthosis, hyperkeratosis, 

parakeratosis, folliculitis and 

epidermatitis. 

Dermal

Intermediate-Term

(1 – 6 months)

and Long-Term

(> 6 months)

No endpoint identified in the toxicity data base for Busan 77 and therefore, this risk assessment 

is not required.

Inhalation 

Short-Term

(1 – 30 Days)

NOAEL = 500 

mg/kg/day

UFA = 10

UFH = 10

UFDB = 10

Target MOE = 1000

Prenatal Developmental Toxicity -

Rats (MRID 41423001/93062018)

LOAEL = 700 mg/kg/day based upon

increased mortality and morbidity 

(convulsions, lethargy and 

pathological changes in lung, brain, 

stomach and intestines). 

Inhalation

Intermediate-Term

(1 – 6 months)

NOAEL = 221 

mg/kg/day

UFA = 10

UFH = 10

UFDB = 10

Target MOE = 1000

Subchronic Oral Toxicity – Rats

(MRID40025001/93662014)

LOAEL = 752 mg/kg/day based upon 

mineralization of renal tubules with 

decreases in body weights, absolute 

organ weights (heart, liver, kidney and 

gonads), equivocal decrease in RBC 

counts, elevated leukocyte counts, 

non-suppurative inflammation in the 

chorid plexus of the brain and 

increased animal death observed at the 

higher doses of 2554 and 3685 

mg/kg/day.

Inhalation 

Long-Term

(> 6 Months)

NOAEL = 100 

mg/kg/day

UFA = 10

UFH = 10

UFDB = 10

Target MOE = 1000

Chronic Toxicity/Carcinogenicity -

Rats

(MRID 41809101/41561301)
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Exposure 

Scenario

Dose Used in Risk 

Assessment 

(mg/kg/day)

Target Margin of Exposure 

(MOE), Uncertainty Factor (UF), 

FQPA Safety Factor (SF)

Study and Toxicological Effects

LOAEL = 300 mg/kg/day based upon 

body weight gain (M), reduced total 

protein (M), reduced inorganic 

phosphorus (M&F), increased urine

pH (F) and a dose related-increase in 

bloody crusts.

Cancer

Busan 77 was classified a Group “D” carcinogen based upon increased thyroid C-cell adenomas 

in rats by the HED RfD Committee (1994). Under registration review, the Agency anticipates 

conducting a carcinogenicity assessment, pending the outcome of review of Busan 77 

carcinogenicity data by the HED CARC.

3.2 Dietary Exposure

3.2.1 Food

The most recent Busan 77 dietary exposure and risk assessment was conducted in the RED in 

2007 for pulp and paper manufacturing uses. The 2007 dietary assessment indicated that there 

were no dietary risks of concern. Based on the current Busan 77 labels, the only use pattern that 

could potentially result in dietary (food) exposures is the use in the manufacture of pulp, paper, 

and paperboard products. There are four labels with pulp, paper, and paperboard uses (57787-23; 

75538-1; 83451-20; 1448-42); however, all of these labels clearly indicate that Busan 77 is not to 

be used in the manufacture of paper products that contact food. As a result, even though Busan 

77 has been cleared as an indirect food additive by the US FDA, a dietary assessment by EPA is 

not required as part of Registration Review since there are no FIFRA-registered food contact 

uses. A dietary assessment will be required if any food-contact pulp, paper, and/or paperboard 

uses are proposed in the future. Since there are no dietary (food) exposures based on the current 

use patterns, food contributions will not be included in the aggregate assessment.

3.2.2 Drinking Water

The Agency plans to conduct a drinking water assessment during this registration review unless 

the expected environmental fate data show low potential for surface water exposure. While 

Busan 77 is a polycationic compound that is expected to sorb tightly to soil, sediment, and 

sludge, the Agency does not have adequate fate data on sorption to determine the potential for 

contamination of drinking water. Busan 77 is resistant to both chemical and microbial 

degradation and to volatility, and therefore sorption is expected to be the primary route of 

dissipation. These fate data are anticipated to be required during registration review.

3.3 Occupational and Residential Exposures

The Agency anticipates the need to revise the occupational and residential assessments 

conducted in support of the 2007 RED. The occupational exposure assessment utilized surrogate 

databases, for which uncertainties have been identified and needs to be updated. Occupational

exposures may occur from liquid pour applications, machinist exposure from metalworking

fluids, metalworking fluids, pulp and paper, and secondary oil recovery in injection water.
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Residential exposures include handler exposures from liquid pour and solid pour from swimming 

pool/spa/whirlpool applications. Residential post-application exposures occur from 

swimming/bathing.

3.3.1 Occupational Handler Exposure

EPA anticipates the need to revise the occupational handler assessment conducted in support of 

the 2007 RED. The Occupational and Residential Exposure (ORE) assessment was conducted on 

July 27, 2007 and is available in the docket to the RED.

3 Occupational handler dermal and 

inhalation exposures to Busan 77 were assessed in the RED for the following applications: liquid 

pour, liquid pump, powders, and machinists. The Agency expects to conduct new assessments 

for liquid pour, solid pour and will also assess machinist exposure for metalworking fluids upon 

receipt of the anticipated exposure data expected from the RED DCI. It should be noted that the 

Agency does not anticipate the need to reassess liquid pump exposures from metered pumps, 

which are now considered minimal exposures. The Agency anticipates that it will need to 

reassess handler exposure for the following use categories: industrial water systems, 

metalworking fluids, pulp and paper, secondary oil recovery in injection water, and swimming 

pools. The Agency expects to conduct new risk assessments upon the receipt of acceptable 

exposure data called in during the RED and also expects to refine exposure assumptions using 

the product use information for registration review. Table 9 lists the scenarios that the Agency

anticipates will need to be reassessed in registration review.

Table 9 – Occupational Handler Exposure Scenarios for Busan 77

Scenario Exposure 

Route(s) Duration

Occupational Exposures

Liquid pour applications for industrial water systems, metalworking 

fluids, pulp and paper, secondary oil recovery in injection water, and 

swimming pools

Dermal 

Inhalation 

Short and 

Intermediate-Term

Machinist exposure to metal working fluids Dermal 

Inhalation 

Short, Intermediate

and Long Term 

3.3.2 Residential Handler Exposures

EPA anticipates the need to revise the residential handler assessment conducted in support of the 

2007 RED. The Agency expects to refine exposure assumptions using the product use 

information for registration review. Residential handler dermal and inhalation exposures to 

Busan 77 were assessed in the RED for swimming pools, and whirlpool/spas. Table 10 lists the 

residential handler scenarios that the Agency anticipates will need to be reassessed in registration 

review.

Table 10 – Residential Handler Exposure Scenarios for Busan 77

Scenario Exposure 

Route(s) Duration

 3 2007 RED Docket Number: EPA-HQ-OPP-2007-0834
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Residential Exposures

Liquid pour swimming pools, spas, and whirlpools Dermal 

Inhalation 

Short and Intermediate 

Term 

Solid place swimming pools, spas, and whirlpools Dermal 

Inhalation

Short and Intermediate 

Term

3.3.3 Residential Post-Application Exposures

EPA anticipates the need to revise the residential post-application assessment conducted in 

support of the 2007 RED. Residential post-application exposures could result when bystanders 

(adults and children) come in contact with Busan 77 in areas where pesticide end-use products have 

recently been applied (e.g., swimming pool and spas/whirlpool ), or when children incidentally ingest 

the pesticide residues through swallowing treated swimming pool water. Note that inhalation 

exposures (vapor) are not expected due to the negligible vapor pressure of Busan 77; therefore, these 

exposures were not assessed. Table 11 lists the residential post-application scenarios that the 

Agency anticipates that will be need to be reassessed in registration review.

Table 11 – Residential Post-Application Exposure Scenarios for Busan 77

Scenario Exposure 

Route(s) Duration

Residential Exposures

Post application exposure to swimming pools and spas/whirlpool Dermal 

Incidental Oral 

Short and Intermediate 

Term 

3.4 Aggregate and Cumulative Exposure

3.4.1 Aggregate Exposures 

The Agency anticipates the need to conduct an aggregate assessment which will include both 

drinking water and residential exposures (i.e. swimming pools/spas). Since there are no dietary 

(food) exposures based on the current use patterns, food contributions will not be included in the 

aggregate assessment.

3.4.2 Cumulative Exposures 

With respect to cumulative exposure, unlike other pesticides for which EPA has followed a 

cumulative risk approach based on a common mechanism of toxicity, EPA has not made a 

common mechanism of toxicity finding as to Busan 77 and Busan 77 does not appear to produce 

a toxic metabolite produced by other substances. For the purposes of this registration review, 

therefore, EPA has not assumed that Busan 77 has a common mechanism of toxicity with other 

substances. For information regarding EPA’s efforts to determine which chemicals have a 

common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see 

http://www.epa.gov/pesticide-science-and-assessing-pesticide-risks/cumulative-assessment-riskpesticides.
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4 Environmental Risk Assessment

The Agency anticipates the need to conduct and ecological risk assessment and may call in data 

in support of the assessment. The ecological risk assessment planned during registration review 

will allow the Agency to determine potential acute and chronic risks to aquatic organisms 

exposed to Busan 77 transported from treatment sites into the aquatic environment. Such sites 

include recirculating cooling towers, air washer systems, petroleum secondary recovery systems, 

and metalworking fluids. There is no evidence of degradation of Busan 77 by either abiotic or 

biotic routes. The primary exposure is expected to be to sediment-dwelling organisms based on 

the sorption potential from the positively-charged parent compound. However, bioconcentration 

is not expected because of the miscibility in water (Table 4).

The Agency has not previously conducted a risk assessment that supports a complete endangered 

species determination for Busan 77. The risk assessment planned for registration review also will 

allow the Agency to determine whether each use of the Busan 77 has 'no effect' or 'may affect' 

federally listed threatened or endangered species (listed species) or their designated critical 

habitats. When an assessment concludes that a pesticide’s use 'may affect' a listed species or its 

designated critical habitat, the Agency will consult with the U.S. Fish and Wildlife Service 

and/or National Marine Fisheries Services (the Services), as appropriate.

4.1 Environmental Fate Assessment

Busan 77 is miscible in water (Table 4) and is not expected to degrade by either abiotic processes 

(MRIDs 41407401 and 41420901) or biotic processes (MRIDs 40334101, 40165201, 40165202).

Movement from water or soil into air is also not expected to occur for most uses based on the 

negligible vapor pressure and Henry’s Law values (Table 4), but the air washer use may have the 

potential to create aerial exposure because the Busan 77 dissolved in water can volatilize with 

the water. Bioconcentration in aquatic organisms is also not expected based on the water 

miscibility and low log P values (Table 4). Sorption to soil, sediment, and sludge is expected to 

be the primary route of dissipation since Busan 77 is a quaternary ammonium compound that has 

a positive electrical charge. However, data for sorption to soil and sludge have not been 

submitted and are anticipated to be required.

Busan 77 has the potential to reach Waste Water Treatment Plants (WWTPs) from the registered 

uses, and data on activated sludge sorption isotherm (ASSI) and activated sludge respiration 

inhibition (ASRI). WWTP biodegradability studies (835.3110, 835.3220, 835.3240, and 

835.3280) are normally required for chemicals that have the potential to reach wastewater 

treatment plants. However, in the case of Busan 77, there is no apparent route of chemical or 

microbial degradation, and therefore degradation studies in a WWTP would not be expected to 

provide additional information and are not anticipated to be required.
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4.1.1 Water Quality

Busan 77 is not identified as a cause of impairment for any water bodies listed as impaired under 

section 303(d) of the Clean Water Act.

4 In addition, no Total Maximum Daily Loads (TMDL) 

have been developed for Busan 77.

5 More information on impaired water bodies and TMDLs 

can be found at EPA’s website.

6

4.2 Conceptual Models for Environmental Exposure Pathways

Based on the summary of registered uses of Busan 77 presented in Table 5, physical/chemical 

properties in Table 4, and environmental fate data presented in Table 4 and Appendix B, the 

Agency has developed conceptual model diagrams for exposure of ecological organisms to 

parent Busan 77. Busan 77 is stable to both abiotic and biotic processes and is unlikely to move 

from soil or water into air based on its negligible vapor pressure and Henry’s law values (Table 

4) with the exception of the air washer use which creates the potential for aerial exposure 

through co-distillation. Chemicals that are released down-the-drain can typically take from a few 

to several hours to reach wastewater treatment plant intakes following their discharge and from 

several hours to roughly a day following their discharge from wastewater treatment plants to

reach surface water. Data on WWTP dissipation have not been submitted for Busan 77, but the 

primary route of dissipation is expected to be sorption to sludge and biodegradation is expected 

to be limited.

The Agency has created conceptual models for potential routes of environmental exposure which 

are included in “Conceptual Models for Environmental Exposure Pathways of Antimicrobial 

Pesticides,” found in the docket at www.regulations.gov, EPA-HQ-OPP-2014-0638.

Use sites and corresponding figures of conceptual model diagrams are as follows:

• Recirculating cooling towers and air washer systems (slides 13 and 14)

• Metalworking fluids (slide 18)

• Paper manufacturing (slide 26)

• Petroleum secondary recovery systems (slides 19 and 20), and 

• Swimming pools, spas, whirlpools, hot tubs, ornamental ponds, aquaria, waterbeds

(Slides 27 and 28)

The screening-level risk assessment that will be conducted as part of this registration review will 

evaluate potential risks to ecological organisms from exposure to parent Busan 77 only because 

it did not degrade by abiotic processes (MRIDs 41407401 and 41420901) or biotic processes 

(MRIDs 40334101, 40165201, 40165202).

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

5

http://iaspub.epa.gov/tmdl_waters10/attains_nation.tmdl_pollutant_detail?p_pollutant_group_id=885&p_pollutant_

group_name=PESTICIDES

6 http://www.epa.gov/owow/tmdl/
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4.3 Ecological Effects Assessment

Previously an ecological risk assessment was not typically conducted for the types of uses 

registered for Busan 77, except for the once-through cooling water use. A risk assessment for 

once-through cooling water uses, using maximum application rates and the toxicity values for 

various organisms, was conducted during the RED (2007) process. The results of the assessment 

of once-through cooling water uses indicate that there may be high risk to freshwater fish, 

freshwater invertebrates and estuarine/marine invertebrates from the use of Busan 77. All other 

uses were considered indoor at the time of the RED assessment and were thought to have 

minimal to no environmental exposure potential following use. During the registration review 

process, all of the current uses of Busan 77 will be evaluated and an updated assessment of the 

once-through cooling water uses will be conducted.

4.3.1 Measures of Effect (Ecotoxicology Endpoints)

Ecotoxicity endpoint data are used as measures of effects to aquatic and terrestrial organisms. All 

available ecotoxicity endpoints in the Agency’s files from chemical studies submitted by 

registrants or other interested parties, federal laboratory data, or from the public literature are 

tabulated in Appendix C along with the data requirements and data gaps. The most sensitive 

values for each receptor group are used for the risk assessment. The selected endpoints from the 

RED assessment (Table 12) will be updated during the risk assessment phase of registration 

review based on any additional submitted data or data found in the public literature. 

Table 12 – Ecological Effects Endpoints Selected for Assessing Risks from Busan 77

Receptor 

Group Surrogate Species Risk Scenario Toxicity Endpoint Reference(MRID)

Freshwater fish

Rainbow trout Acute 96-h LC50 = 0.047 mg/L 41352001

Fathead minnow Chronic NOAEC = 0.019 mg/L 49362401

Freshwater 

invertebrates Daphnia magna Acute 48-h EC50 = 0.28 mg/L 41352003

Chronic Data Gap --

Estuarine/marine fish Sheepshead minnow

Acute 96-h LC50 = >360 mg/L 40139202

Chronic Not required1 --

Estuarine/marine 

invertebrates

Quahog clam Acute 48-hr EC50 = 0.21 mg/L 40334201

Mysid Shrimp Acute 96-hr LC50 = 7.8 mg/L 40139203

Chronic Not required1 --

Algae Green alga Non-listed 96-hr EC50 = 0.003 mg/L 49453501

Aquatic vascular 

plants Duckweed Non-listed Data Gap2 --

Seedling Emergence Oat Non-listed EC25 = >73 mg/L 42038101

Vegetative vigor Corn Non-listed EC25 = >74 mg/L 42038101

Birds

Mallard duck Acute LD50 = 497 mg/kg 41654801

Bobwhite quail Subacute 

dietary

LC50 = >3462 mg/kg 00159307

Chronic Not required1 --

Mammals Acute LD50 = 1951 mg/kg (males)

LD50 = 2587 mg/kg (females) 41373401
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Receptor 

Group Surrogate Species Risk Scenario Toxicity Endpoint Reference(MRID)

Chronic NOAEL = 100 mg/kg/day 41234501

Nontarget insects Honey bee Acute Not required1 --

1 Not required for this use pattern.

2 Submitted study 42013301 is supplemental and did not provide an endpoint.

4.4 Exposure Analysis Plan

4.4.1 Aquatic and Terrestrial Wildlife Exposure Estimates

Available OPP models will be used to determine the estimated environmental concentration 

(EECs) in the aquatic environment. Resulting exposure estimates will be compared to the 

toxicity endpoints to determine if levels of concern are exceeded for each receptor group. 

4.4.2 Screening Level Down-the-Drain Analysis

A screening Down-the-Drain (DtD) assessment was not conducted for Busan 77 and degradates.

Since Busan 77 is expected to sorb strongly to soils, but no quantitative sorption data are 

available, risk estimates from the screen would have little value. 

Further, the DtD screen only pertains to domestic wastewater treatment plants. Based on 

registered uses of Busan 77 presented in Table 5 of this FWP, there are some uses, such as 

commercial and industrial cooling water systems and paper mill water systems, for which 

discharges may enter industrial WWTPs rather than domestic WWTPs prior to entering surface 

water. Different methods and tools for assessing potential exposures and associated ecological 

risks will be applied for these other types of uses.

The Agency does not have high quality data on the toxicity of Busan 77 to activated sludge 

microorganisms in wastewater treatment plants or data on sorption to sludge. Consequently, the 

Agency anticipates needing data on Busan 77 to fulfill data gaps regarding toxicity to activated 

sludge microorganisms and during wastewater treatment.

4.5 Effects Analysis Plan

Additional open literature studies will be identified through EPA’s ECOTOXicology (ECOTOX) 

database. The ECOTOX database will be searched during the risk assessment phase of Busan 77 

and the data reviewed following the Agency’s policy for reviewing public literature for use in

assessing risks from pesticides.

7 Those ecotoxicity values found in the public literature to be 

more sensitive than those currently in Table 12 and or which satisfy an existing data gap(s) will 

be used during the risk assessment phase of Busan 77 to update Table 12. ECOTOX was created 

 7

http://www.epa.gov/pesticides/science/efed/policy_guidance/team_authors/endangered_species_reregistration_wor

kgroup/esa_evaluation_open_literature.htm
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and is maintained by the USEPA, Office of Research and Development, and the National Health 

and Environmental Effects Research Laboratory's Mid-Continent Ecology Division.

5 Endocrine Disruptor Screening Program (EDSP)

As required by FIFRA and FFDCA, EPA reviews numerous studies to assess potential adverse 

outcomes from exposure to chemicals. Collectively, these studies include acute, subchronic and 

chronic toxicity, including assessments of carcinogenicity, neurotoxicity, developmental, 

reproductive, and general or systemic toxicity. These studies include endpoints which may be 

susceptible to endocrine influence, including effects on endocrine target organ histopathology, 

organ weights, estrus cyclicity, sexual maturation, fertility, pregnancy rates, reproductive loss, 

and sex ratios in offspring. For ecological hazard assessments, EPA evaluates acute tests and 

chronic studies that assess growth, developmental and reproductive effects in different 

taxonomic groups. As part of its reregistration decision for Busan 77, EPA reviewed these data 

and selected the most sensitive endpoints for relevant risk assessment scenarios from the existing 

hazard database. However, as required by FFDCA Section 408(p), Busan 77 is subject to the 

endocrine screening part of the Endocrine Disruptor Screening Program (EDSP). 

EPA has developed the EDSP to determine whether certain substances (including pesticide 

active and other ingredients) may have an effect in humans or wildlife similar to an effect 

produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator 

may designate.” The EDSP employs a two-tiered approach to making the statutorily required 

determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a 

chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal 

systems. Chemicals that go through Tier 1 screening and are found to have the potential to 

interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA 

will determine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2 

testing is designed to identify any adverse endocrine-related effects caused by the substance, and 

establish a dose-response relationship between the dose and the E, A, or T effect. 

Under FFDCA Section 408(p), the Agency must screen all pesticide chemicals. Between October 

2009 and February 2010, EPA issued test orders/data call-ins for the first group of 67 chemicals, 

which contains 58 pesticide active ingredients and 9 inert ingredients. A second list of chemicals 

identified for EDSP screening was published on June 14, 20138 and includes some pesticides 

scheduled for registration review and chemicals found in water. Neither of these lists should be 

construed as a list of known or likely endocrine disruptors.

For further information on the status of the EDSP, the policies and procedures, the lists of 

chemicals, future lists, the test guidelines and the Tier 1 screening battery, please visit our 

website.9

 8 See http://www.regulations.gov/#!documentDetail;D=EPA-HQ-OPPT-2009-0477-0074 for the final second list of 

chemicals. 9 http://www.epa.gov/endocrine-disruption
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6 Optional Label Changes

The Agency invites any label amendments that could be considered to eliminate the anticipated 

need for EPA to require certain data, reduce the possibility that EPA’s planned risk assessments 

overestimate risk due to reliance on conservative assumptions, and/or improve label clarity.

7 Next Steps

A DCI will be developed requiring generation and submission of the data listed under the 

“Anticipated Data Needs” Section of this document. The Agency expects to issue the DCI by 

March of 2017.
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8 References 

MRID 42372401. Bowles, D. (1991) Product Chemistry for WSCP: Lab Project Number: NA 

909218/1. Unpublished study prepared by Twin City Testing Corp. and NATEC Institut 

fur Naturwissenschaftlich-technische Dienste GmbH. 88 p.

MRID 46915602. Wo, C. (2006) Physical and Chemical Characteristics (830 Series, Group B): 

Algae Clean Out W. Unpublished study prepared by Product Safety Laboratories. 6 p.

MRID 47120304. Tillman, A. (2007) Waiver Request for Certain Data Requirements for WSCP 

MUP. Project Number: KT/200704. Unpublished study prepared by Kerley Trading, Inc. 

7 p.

MRID 49176301. Dotse, A.; Grabowicz, J. (2013) Product Chemistry for WSKT: Active 

Ingredient and Physical/ Chemical Properties: Final Report. Unpublished study prepared 

by Buckman Laboratories, Inc. 27p.

U.S. Environmental Protection Agency (EPA). 2002. ECOTOX User Guide: ECOTOXicology 

Database System. Version 3.0. Available: http://www.epa.gov/ecotox/

U.S. Environmental Protection Agency. High Production Volume Information System (HPVIS). 

http://www.epa.gov/hpvis/

U.S. Environmental Protection Agency (US EPA). 1999. Evaluation of Chemical

Manufacturers Association Antimicrobial Exposure Assessment Study. Memorandum 

from Siroos Mostaghimi, Ph.D., USEPA, to Julie Fairfax.
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Appendix A Toxicology Profile

Acute Toxicity for Product Labeling 

The acute toxicity database for Busan 77 is considered complete and testing results are 

summarized in Table A1. Busan 77 tested at 60-61.6% active ingredient has a moderate order of 

acute toxicity via the oral, dermal and inhalation routes of exposure (Toxicity Category III). For 

dermal irritation, Busan 77 has a low order of acute toxicity (Toxicity Category IV) and is a 

slight eye irritant (Toxicity Category IV). Busan 77 is not a dermal sensitizer.

Table A1 – Acute Toxicity Studies for Busan 77 (60 – 61.6% active ingredient)

Guideline No./

Study Type MRID No. Results Toxicity Category

870.1100/ 

Acute oral toxicity 

41373401

93062009

LD50 = 1951 (1727-2203) mg/kg )M)

LD50 = 2587 (2059-3250) mg/kg (F) III

870.1200/ 

Acute dermal toxicity 

41327101

93062010 LD50 > 2000 mg/kg III

870.1300/ 

Acute inhalation 

toxicity 

41877501

LC50 = 4.0 (2.3-7.1) mg/L (M)

LC50 = 2.4 (1.7-3.3) mg/L (F)

LC50 = 2.9 (2.3-3.7) mg/L (combined)

III

870.2400/ 

Acute eye irritation 

41361701

93062011 Redness, cleared on day 3 post-application III

870.2500/ 

Acute dermal irritation 

41298601

93062012 Slight irritant IV

870.2600/ 

Skin sensitization 

40750301

93062013 Not a sensitizer N/A

N/A=Not applicable; M = males; F = Females

Subchronic Toxicity 

Subchronic toxicity studies (MRIDs 40170601/93062015 and 40025001/93662014) indicate that 

the dermal route did not result in systemic effects when tested to the limit dose (1,000 

mg/kg/day). However, signs of chronic irritation/inflammation were noted in both males and 

females at the 100 and 1,000 mg/kg/day dose level (skin lesions included ulceration of the 

epidermis, chronic inflammation of the edermis, acanthosis, hyperkeratosis, parakeratosis, 

folliculitis, and epidermatitis). The oral route led to toxic effects at 1,000 mg/kg/day 

(mineralization of renal tubules).

Developmental Toxicity 

Developmental toxicity studies (MRIDs 41423001/93062018 and 41248001/93062019) through 

the oral route showed developmental effects occurred at greater than or equal dose levels as 

maternal effects. A study conducted in rats had maternal effects occurring at 350 mg/kg/day 

(decreased food consumption and body weight gain, which were not statistically significant) and 

became severe at 700 mg/kg/day (mortality, morbidity observed, including lungs, brain, stomach, 
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and intestines). No developmental effects were observed in the doses tested in this study. In a 

study in rabbits, maternal effects (increased incidence of abortions, clinical signs including 

reduced defecations and emaciation, significantly reduced feed consumption and body weight 

gain) were seen at the same dose, 45 mg/kg/day, as developmental effects (increased incidence 

of ribs and unossified sternabrae).

Reproductive Toxicity

In a reproduction study in rats (MRID 40578201/93062020), systemic effects (reduced body 

weight, reduced absolute and relative liver weight, inflammation of the choroid plexus in the 

brain, and tubular mineralization of the kidney were seen at doses ~300 mg/kg/day and above) 

were seen at lower doses than reproductive effects (decrease in number of live pups on days 0 

and 4 after birth in both generations at ~600 mg/kg/day) in rats.

Chronic Toxicity 

A chronic toxicity study in dogs (MRID 41234501/93062016) by the oral route showed signs of 

toxicity in males (testicular hypoplasia, atrophy/degeneration, aspermia, dysplasia and cellular 

debris of testicular origin in the epididymis at 500 mg/kg) at lower doses than females (G-I 

disturbances, emaciation and nervous symptoms at 1,000 mg/kg).

Two chronic, feeding studies were conducted with Busan 77; a combined chronic 

toxicity/carcinogenicity study in rats (MRID 41809101/41561301) and a chronic toxicity study 

in mice (MRID 41494301/93062017). For additional chronic toxicity information, see 

“carcinogenicity” section below.

Carcinogenicity 

In a combined chronic/carcinogenicity study (MRID 41809101/41561301), rats exposed to 0, 

100, 300 or 900 mg/kg/day (0, 2000, 6000 and 18000 ppm in males and females) a chronic 

NOAEL of 100 and LOAEL of 300 mg/kg/day, was based on reduced body weight gain (F),

reduced albumin (CM), reduced total protein (CM), reduced phosphorus (inorganic) (M & F),

increased urine pH (F) and a dose-related increase in bloody crusts (M). In addition, at 900 

mg/kg/day, hyperesthesia, bloody crusts, decreased body weight gain, decreased phosphorus 

(inorganic), globulin and total protein and increased brain-to-body weight ratio associated with 

mineralization of the brain in the thalamic region, in males and females; decreased albumin, 

glucose and cholesterol in males and increased urine pH in females, were observed. In the 

carcinogenicity study, for females, there was a possible increased incidence of thyroid C-cell 

adenomas in the 300 and 900 mg/kg/day groups and a positive trend in males and females.

In a carcinogenicity study in mice (MRID 41494301/93062017), there was a dose related 

increase in proteinaceous casts in the renal tubules of males and renal pelvic and tubular

dilatation in males and females. It could not be definitively determined whether increased 

incidences of rough hair coat, bloody crusts and red ears in males and convulsions in females 

occurred at the low or high dose, 600 or 2400 mg/kg/day, respectively. Busan 77 in the diet was 

determined to be non-carcinogenic in mice up to 2,400 mg/kg/day. 
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Carcinogenic Potential 

There was an increased incidence of thyroid C-cell adenomas in females at the mid- and highdose levels and in males at the high dose level in the combined chronic toxicity/carcinogenicity 

in rats. The increase in the overall incidence of thyroid C-cell adenomas in males did not attain a 

statistically significant level when compared with the concurrent controls. However, there was a 

statistically significant positive dose-related trend in males and females for this type of tumor.

The incidence of C-cell adenomas in females was outside the range and higher than the mean of 

historical control incidences. Historical control incidences for this type of tumor in this strain of 

rat were as follows: a) in females, the range was 0- 14% with a mean value of 4.5%, b) in males, 

the range was 1.7-11% with a mean value of 5.8%. Only one C-cell carcinoma (1/24; 4.1%) was 

observed in the mid-dose males. The increase in tumor incidences in rats might have been due to 

alteration of the normal metabolic pathways, biotransformation and/or elimination of the 

chemical as a result of compromised liver and kidney function. In contrast, Busan 77 did not 

induce tumors in Crl:CD-1 (ICR) BR mice up to 16,000 ppm (2,400 mg/kg/day) in the diet and 

therefore, is not carcinogenic. Consequently, Busan 77 was classified a “Group D” carcinogen 

based upon increased thyroid C-cell adenomas in rats by the HED RfD Committee (1994).Under 

registration review, the Agency anticipates conducting a carcinogenicity assessment, pending the 

outcome of a review of Busan 77 carcinogenicity data by the HED CARC.

Mutagenicity 

The mutagenicity battery previously submitted for Busan 77 showed no evidence of 

mutagenicity or genotoxicity. Busan 77 was not mutagenic in a reverse mutation assay in 

bacteria in the presence or absence of mammalian metabolic activation (MRID 41573701), 

produced no increase in sex-linked recessive lethal genes in Drosophila flies (MRID 00151205), 

did not increase micronuclei in mice (MRID 00151206/93062021) and did not increase 

unscheduled DNA synthesis in primary rat hepatocytes (MRID 40978701/93062022). 

However, this mutagenicity battery is now considered incomplete. Under current standards, this 

data gap can be fulfilled via conduct of either an in vitro mammalian cell gene mutation assay 

(OCSPP 870.5300) or an in vitro mammalian chromosome aberration test (OCSPP 870.5375) 

with the Busan 77 technical grade active ingredient.

Other Toxicological Effects 

Neurotoxicity

There was evidence of neurological effects in the existing toxicity database for Busan 77.

Convulsions were observed in the mouse carcinogenicity study (MRID 41494301/93062017) 

with Busan 77 at a dose of 8000 ppm (~1200 mg/kg) in males (10/50 mice vs. 6/50 mice in 

controls). In females, there appeared to be increased incidence of convulsions at 4000 ppm (~600 

mg/kg) (7/50 mice vs. 3/50 mice in controls). However, the time of appearance and duration of 

this clinical sign was not established in this study. In a chronic toxicity/carcinogenicity study in 
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rats (MRID 41809101/41561301), mineralization of the thalamic region of the brain was 

observed at a dose of 900 mg/kg/day at week 105 as was an increased incidence of hyperplasia in 

males and females at this dose (36/60 males; 21/60 females vs. 0/60 controls for both sexes). A 

developmental toxicity study in rats (MRID 41423001/93062018) showed dilated meningeal 

vessels and/or hemorrhage in maternal rats at a dose of 700 mg/kg/day. In a two-generation 

reproduction toxicity study (MRID 40578201/93062020), increased incidence of inflammation 

of the choroid plexus of the brain was observed in parental animals of both generations at doses 

of 300 mg/kg/day and above. Based on this evidence, potential neurotoxic effects can occur in 

humans exposed to Busan 77. Therefore, a neurotoxicity battery in rats (OCSPP 870.6200) that 

includes both acute and subchronic (90-day) neurotoxicity screening studies are anticipated to be 

required for Busan 77.

Metabolism and Pharmacokinetics

Metabolism of Busan 77 (MRID 40268601/93062024) showed no expired 14CO2. Under 

intravenous injection, roughly half (52-55%) of the radioactivity was excreted after 7 days 

through urine (38-44%) and feces (11-14%), while tissues contained 43-55% of the dose. The 

major route of excretion after single oral and repeated oral doses was feces (85-103%), urine was 

minor (3%). Of the oral dose groups the highest residual radioactivity was found in kidneys, liver 

and spleen. Recoveries of doses in the tissues, including carcass (≤0.14%) indicated a minimal 

potential for bioaccumulation after oral dosing.

Dermal Penetration

In a dermal penetration study (MRID 40139201/93062025), 14C-Busan 77 (purity 60%), was 

administered dermally to male CRL:CD BR rats(24/group) at single dose levels of 0, 2, 20 or 

300 mg/rat, (0.16, 1.59 or 15.97 mg/cm2

, respectively). Less than 0.2% of the 14C-Busan 77 was 

absorbed. Mean % recovery of radioactive dose ranged from 74.5 - 92.7%. The majority of the 

dose was recovered in the skin rinse (65.5% - 88.6%) and at the skin site (0.4% - 13.7%).

Determination of Special Susceptibility

There is an adequate database for determining potential sensitivity of Busan 77 to offspring. Two 

developmental toxicity studies conducted with rats and rabbits demonstrated developmental 

toxicity effect levels that were at the same or higher dose levels than those doses causing 

maternal toxicity. In the rat, a developmental toxicity LOAEL was not established; the NOAEL 

was 500 mg/kg/day. The maternal LOAEL of 350 mg/kg/day in this study was based on 

decreased body weight gain and food consumption. In rabbits, the maternal and developmental 

toxicity LOAELs were the same, 45 mg/kg/day. Maternal effects consisted of increased 

abortions, reduced defecation and emaciation, reduced food consumption, and decreased weight 

gain. Offspring effects at this dose consisted of increased incidence of 13th rudimentary ribs and 

unossified sternebrae #5 and #6. In the two-generation reproduction toxicity study, the 

reproductive LOAEL was >900 mg/kg/day, whereas the parental LOAEL was 600 mg/kg/day.

The weight of evidence from these data indicates no increased sensitivity of the offspring in 

either the developmental or reproductive toxicity studies to the adverse effects of Busan 77. This 
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conclusion is supported by the previous conclusion of the HED RfD/Peer Review Committee in 

1994.

FQPA Considerations

The FQPA Safety Factor (as required by the Food Quality Protection Act of 1996) is intended to 

provide an additional 10-fold safety factor (10X), to protect for special sensitivity in infants and 

children to specific pesticide residues in food, drinking water, or residential exposures, or to 

compensate for an incomplete database. The Antimicrobials Division’s Toxicology Endpoint 

Selection Committee (ADTC) has recommended that the special hazard-based FQPA safety factor 

for Busan 77 be removed. This conclusion is based upon the (1) availability of acceptable 

developmental and reproductive toxicity studies conducted with Busan 77 that adequately 

characterize the dose-response of this compound (2) the lack of evidence for any sensitivity of 

fetuses or offspring to the adverse effects of Busan 77 and (3) the risk assessment does not 

underestimate the potential exposure for infants and children.
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Toxicity Profile for Busan 77

Table A2 – Subchronic, Chronic, and other Toxicity Profiles for Busan 77

Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

870.3050

28-day oral (mice)

Purity: not reported

MRID 40362601

Kehoe, D.F. (1987). Four-Week Toxicity Study with Busan 77 

in Mice. Buckman Labs, Inc., Memphis, TN. Study #: 6176-

108

Not reviewed

Busan 77 administered orally in the diet at 0, 2500, 5000, 

10000, 20000 ppm for 28 days

5 rats/sex/dose

NOAEL = 10,000 ppm

LOAEL = 20,000 ppm 

Body weights, food consumptions, and ante mortem observations were 

recorded at initiation and then weekly for at least 4 weeks. At the end 

of the study, all surviving mice were bled for hematology and clinical 

chemistry tests. The mice were then anesthetized, exsanguinated, and 

necropsied. A female mouse that died on test was also necropsied and 

representative tissues were collected. At the scheduled necropsies, 

macroscopic observations were recorded, selected organs were 

weighed, and representative tissues were collected a; required by the 

protocol. Specified tissues from mice given 0 or 20,000 ppm were 

examined microscopically. Representative samples of gross lesions and 

rectums were also examined microscopically from all groups.

No treatment-related ante mortem observations were seen in animals 

fed Busan 77; and no treatment-related changes occurred in body 

weight or food consumption the Cumulative body weight gains were 

significantly lower during Week 1, 2 and 4 in males fed 20,000 ppm 

Busan 77. Although females fed 20,000 ppm had a lower body weight 

when compared with the control group, there were no statistically 

significant differences.

There were no treatment-related effects on the clinical pathology data, 

organ weight data, or microscopic observations. The only microscopic 

lesions that may have been treatment related were small foci of 

neutrophils at the anorectal junction of mice given 20,000 ppm Busan 

77.

870.3100

90-day oral (rat)

Purity: not reported

MRID 40025001/93662014

Tisdel, Merrill (1986). Thirteen Week Subchronic Toxicity 

Study With Busan 77 in Rats. Buckman Labs, Inc. Memphis, 

TN. Study #: 6176-106.

Core-Minimum

NOAEL = 3,000 ppm(2000 mg/kg) 

LOAEL = 10,000 ppm (752 mg/kg) based on mineralization of renal 

tubules. At 30,000 ppm and 40,000 ppm (approx. 2554 and 3685 

mg/kg/d, resp.) the following were observed: decreases in body 

weights and possibly absolute organ weights (heart, liver, kidney and 

gonads); an equivocal decrease in red blood cell counts; elevated 

leukocyte counts; non-suppurative inflammation in the choroid plexus 

of the brain; death.
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

Busan 77 administered orally in the diet at 0, 3000, 10,000, 

30,000 and 40,000ppm (0, 300 1000, 3000, and 4000 mg/kg ) 

for 13 weeks

10 rats/sex/dose

The occurrence of increased kidney weight and mineralization at 

10,000 ppm and above suggests a treatment—related response. 

Severity of mineralization was not discussed. Chronic inflammation of 

the choroid plexus was observed at the two highest dose levels only, 

suggesting a treatment—related response in this parameter. The 

decrease in actual numbers of animals affected at the highest dose level 

(40,000 ppm) is a function of the mortality that was observed at that 

level (8 males, 2 females).

870.3250

90-day dermal (rabbit)

Purity: 60%

MRID 40170601/93062015

Spindler, M. (1987). 13-Week Dermal Toxicity Study with 

Busan 77 in Rabbits. Buckman Laboratories, Inc. Memphis, 

TN.

Study #: 6176-118

Acceptable/Guideline

Busan 77 applied dermally at 0, 10, 100, 1000 mg/kg for 13 

weeks

10 rabbits/sex/dose

Systemic Toxicity

NOAEL > 1000 mg/kg/day (HDT)

LOAEL = Not established

Dermal Toxicity

NOAEL = 10 mg/kg/day

LOAEL = 100 mg/kg/day based on treatment related dermatological 

changes consistent with chronic irritation/inflammation. The skin 

lesions consisted of one or more combinations of ulceration of the 

epidermis, chronic inflammation of the dermis, acanthosis, 

hyperkeratosis, parakeratosis, folliculitis and epidermatitis. 

Following the dermal application of Busan 77 to male and female 

rabbits at dose levels of 0, 10, 100 and 1,000 mg/kg/day, 6 hours/day, 5 

days/week for 13 weeks, there were no treatment-related changes in the 

body weights, food consumption, hematology, clinical chemistry, 

organ weights and organ—to-body weight ratios and gross and/or 

histopathological changes of internal organs of males and females. 

However, there was treatment related dermatological changes 

consistent with chronic irritation/inflammation were seen in males and 

females receiving 100 and 1,000 mg/kg/day. The skin lesions consisted 

of one or more combinations of ulceration of the epidermis, chronic 

inflammation of the dermis, acanthosis, hyperkeratosis, parakeratosis, 

folliculitis and

epidermatitis. The doses tested did not establish the MTD, however, 

the highest dose tested reached Limit Dose.

870.3700

Developmental Toxicity – oral gavage 

(rat)

Purity: 97%

MRID 41423001/93062018

Nemac, Marc D. (1987). A Teratology Study in Rats with 

Busan 77. Buckman Laboratories, Inc. Study #: WIL-94020.

Core-Minimum/Guideline

Maternal Toxicity

NOAEL = 70 mg/kg/day 

LOAEL = 350 mg/kg/day, based on decreased food consumption and 

body weight gain. Mortality observed at 700 mg/kg/day
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

Busan 77 administered orally at 0, 70, 350 500 mg/kg/day on 

GD 6-15

25 rats/dose group

Developmental Toxicity

NOAEL = 500 mg/kg/day 

LOAEL > 500 mg/kg/day (HDT)

Busan 77 was toxic to rat dams, causing decreases in food consumption 

and (non-statistically significant) decreases in body weight gains at 350 

mg/kg/d and 500 mg/kg/d, and death and morbidity at 700 mg/kg/d. 

The dose response curve appears to be inordinately steep, since 1) none 

of the toxicity (convulsions, lethargy and pathological changes in lung, 

brain, stomach and intestines) observed at 700 mg/kg/d was observed 

at 500 mg/kg/d, which is only ‘/, of a log dose lower; and 2) the profile 

of other adverse effects such as decrease food consumption (Table 2) 

suggests a more gradual slope. However, absence of effects at lower 

dose levels cannot be adequately verified from the data submitted with 

this study (see Deficiencies Section above). It is also remarkable that 

liver and/or kidney effects were not reported as being observed, even at 

levels that killed 25% of the dams.

Busan 77 appears to cause a dose-related increase in embryonal death, 

as manifested by a statistically significant increase In early resorptions, 

at 500 mg/kg/d.

870.3700

Developmental Toxicity – oral gavage 

(rabbit)

Purity: 60.8%

MRID 41248001/93062019

Rodwell, Dean E. (1989). Teratology Study in Rabbits with 

Busan 77. Buckman Laboratories, Inc. Study #: 3138.29

Core/Guideline

Busan 77 administered via gavage at 0, 15, 45 and 125 mg/kg 

on GD 6-18 inclusive

20 rabbis/dose group

Maternal Toxicity

NOAEL = 45 mg/kg/day 

LOAEL = 125 mg/kg/day, based upon increased incidence of 

abortions, clinical signs including reduced defecation and emaciation, 

significantly reduced feed consumption and body weight gain during 

treatment.

Developmental Toxicity

NOAEL = 45 mg/kg/day 

LOAEL = 125 mg/kg/day

Based upon increased incidence of 13th rudimentary ribs and 

unossified sternebrae #5 and #6.

The data reporting was thorough and the summary means that were 

validated were supported by the individual animal data, except for 

minor discrepancy in reporting the mean body weight gain of 80 

instead of 183 for the high dose dams during gestation Day 24 - 29 

(Report summary Table 4). The error had been factored in, in the 
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

calculation of body weight gain of dams and will not alter the 

conclusions of the study.

It was reported that the doses were selected based upon a range finding 

study, but no details were provided. While this information should 

have been included in the report, the deficiency is not a factor since 

results of current study indicate maternal toxicity at 125 mg/kg/day.

a. Maternal Toxicity: At 125 mg/kg/day, increased incidence of 

abortions, clinical signs including reduced defecation and emaciation, 

significantly reduced feed consumption and body weight gain occurred 

during treatment.

b. Developmental Toxicity: At 125 mg/kg/day, the incidence of 13th 

rudimentary ribs and unossified sternebrae #5 and #6 was increased 

significantly (P 0.05). The litter incidences for 13th rudimentary rib 

(93.3 vs. 89.5 of historical controls) and unossified sternebrae #5 

and/or #6 (40.0 vs. 36.8 of historical controls) are slightly above the 

corresponding historical controls values. Although, the incidence 

lacked a trend or dose- relationship and occurred at a maternally toxic 

dose, the increases are considered treatment-related. 

870.3800

Reproduction and Fertility Effects (rat)

Purity: not reported

MRID 40578201/93062020

MacKenzie, K.M. (1988). Two-Generation Reproduction 

Study with Busan 77 in Rats (One Litter per Generation).

Buckman Laboratories, Inc., Memphis, TN. Study #: HLA 

6176-104.

Core-Minimum/Guideline

Busan 77 administered in the diet at 0, 6000, 12000 and 18000 

(0, 300, 600 and 900 mg/kg/day) for two generations

25 rats/sex/dose

Maternal Toxicity

NOAEL: not established

LOAEL: 300 mg/kg/day, based on decreased body weight, 

inflammation of choroid plexus, tubular mineralization of the kidney.

Reproductive Toxicity:

NOAEL = 600 mg/kg/day

LOAEL = 900 mg/kg/day based on decreased live pups.
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

870.4100

Chronic Toxicity (dog)

Purity: not reported

MRID 41234501/93062016

Kehoe, Daniel F. (1989). One-Year Chronic Toxicity Study 

with Busan 77 in Dogs. Buckman Laboratories, Inc. Memphis, 

TN. Study #: HLA 6176-111

Supplementary

Busan 77 administered orally at 0, 10000, 20000 and 40000 

ppm (0, 250, 500, 1000 mg/kg) for 52 weeks

4 dogs/sex/dose

NOAEL: 250 mg/kg/day (M); 500 mg/kg/day (F)

LOAEL = 500 mg/kg/day (M) based on testicular hypoplasia, 

atrophy/degeneration, aspermia, dysplasia and cellular debris of 

testicular origin in epididymis.

LOAEL = 1,000 mg/kg/day (F) based on G-I disturbances, weight 

loss and nervous symptoms (bloody stools, emaciation and ataxia, 

respectively).

Administration of Busan 77 in the feed for up to 52 weeks had no 

effect on mortality. However, an increased incidence of vomits and 

abnormal stools (soft and bloody) was observed with increasing dose. 

In the KTD group, all males and females exhibited bloody stools at 12 

(1) and 26 (2) weeks, respectively. Probably this is related to treatment. 

At necropsy, thickened wall of duodenum (3 males and 2 females), 

jejunum (2 males) and ileum (1 male) were observed in the 40,000 ppm 

group. Microscopic examination of these tissues did not reveal any 

changes associated with the treatment and is probably of no 

toxicological significance. In addition, microscopic examination of 

rectum or colon of all HDT males revealed changes associated with 

chronic inflammatory processes and was probably treatment related. 

Body weight gain of males and females in the 40,000 ppm group 

decreased significantly when compared to controls. Body weight gain 

of males and females was reduced by 39 and 42%, respectively, at 12 

weeks when compared to controls. In the 20,000 ppm group, the mean 

body weight gain decreased non-significantly in males and females 17 

and 6%, respectively, at 12 weeks when compared to controls; and 

decreased in males and females 30 and 14%, respectively, at 

termination (52 weeks) when compared to controls and it may not be 

related to treatment. Food consumption was significantly decreased in 

the males and females in the 40,000 ppm group at several intervals. At 

week 13, food consumption of males and females in the 40,000 ppm 

group decreased 46 and 53%, respectively, when compared to controls.

At 26 weeks, males and females in the 40,000 ppm group had a 

significant increase in Hct, MCV, MCH and platelets, and in females a 

significant increase in Hb was observed. Also, a significant decrease in 

RBC was seen in males and females at the 26 week time interval. The 

low RBC and increased MCV may signal mild anemia; however, 
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

histopathology of bone marrow smears and bone sections did not 

correspond to morphological changes. These changes in the HTD 

group may be in part/whole related to mild anemia, emaciation and 

dehydration. Significant increases in MCV in the 10,000 and 20,000 

groups in males/females are of questionable biological significance 

because of the sporadic nature of occurrence.

Total protein, albumin and creatinine were significantly decreased in 

males and females in the 40,000 ppm group at 26 weeks, probably due 

to anemia and emaciation and may be treatment related. Also, a 

significant decrease in K, glucose in males and Phosphorus (inorganic), 

Na and Cl in females were also observed at this interval. This is 

probably due to dehydration, associated with vomition/diarrhea. 

Changes noticed in the 10,000 ppm group and in the 20,000 ppm group 

for K, and Phosphorus (inorganic) at 26 weeks, are of sporadic nature 

and have no toxicological significance. The significant (slight) increase 

of SGPT in males at 26 weeks, in the 40,000 ppm group and of SCOT 

at 52 weeks, in the 10,000 and in the 20,000 ppm groups at 52 weeks, 

is probably of no toxicological significance, since the histopathology 

revealed no treatment related affects. At 52 weeks, males and females 

had significantly reduced cholesterol in the 20,000 ppm group; the 

toxicological significance of this is uncertain.

Urine pH in males was significantly reduced in the 40,000 ppm group 

at 26 weeks and in the 10,000 and in the 20,000 ppm groups at 52 

weeks. Also, higher urinary blood pigment was noticed in the 40,000 

ppm group. The significance of low urine pH and higher urinary blood 

pigment on the urinary tract is of questionable values, since there was 

no corresponding histopathological changes.

Organ to body weight ratios for brain, liver and kidney increased 

(significantly) 52, 19 and 47%, respectively for males in the 20,000 

ppm group at termination, when compared to controls. The actual 

weights of the brain, liver, and kidney in males were 1, -18 and -13 %, 

respectively, when compared to controls. Females exhibited significant 

increases of 4 and 47 %, respectively, in the body weight ratio of 

kidney and liver. The apparent increase in the organ to body weight 

ratios, and actual decrease in the absolute organ weights without 

corresponding histopathological changes, suggest that the changes seen 

grossly may be due to in part/whole to body weight loss, especially 
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

water loss from vomiting and diarrhea. In addition, testes relative organ 

weights were not affected, whereas actual weights decreased 27, 20 

and 58 % in the LTD, MTD, and NW, respectively, when compared to 

controls. Reduced testicular weights corresponded to testicular 

hypoplasia on histopathology, evidenced by atrophy/degeneration, 

aspermia, dysplasia and presence of cellular debris of testicular origin 

in the epididymis of all males in the 40,000 ppm group and 2 males in 

the 20,000 ppm group.

870.4200

Carcinogenicity (mice)

Purity: 60.8%

MRID 41494301/93062017

Kehoe, Daniel F. (1990). 18 Month Study Chronic Toxicity 

Study with Busan 77 in Mice. Buckman Laboratories, Inc. 

Memphis, TN. Study #: HLA 6176-109

Core-Minimum/Guideline

Busan 77 administered orally at 0, 4000, 8000 and 16000 ppm 

(0, 600, 1200, and 2400 mg/kg)for 18 months

50 mice/sex/dose

NOAEL= not established

LOAEL <600 mg/kg/day, based on enlarged kidney pelvis, diffuse 

rough kidney I females; dose-related increase in proteinaceous casts in 

the renal tubules of males, and renal pelvic and tubular dilation in 

males and females.

It could not be definitively determined whether increase incidence of 

rough hair coat, bloody crusts and red ears in males and convulsions in 

females had a LEL of 4,000 or 16,000 ppm.

At the 8000 and above, all of the above findings and including 

convulsions (M & F), urine scams (H & F), rough hair coat (F), bloody 

crusts (F), swollen abdomen (M & F), swollen anus (M), red tails (H & 

F), red ears (F), reduced body weight gains (H & F), diffuse rough 

kidneys (H & F), large kidney pelvis (H & F), relative kidney weights 

(high dose males, and mid and high dose females), proteinaceous casts 

(F), cortical fibrosis/scarring (F) and cysts (F), were observed.

870.4300

Combined chronic 

toxicity/carcinogenicity (rat)

Purity: 60%

MRID 41809101/41561301

Kehoe, Daniel F. (1991). Combined Chronic Toxicity Study 

and Carcinogenicity Study with Busan 77. Buckman 

Laboratories, Inc. Memphis, TN. Study #: HLA 6176-107.

Core-Minimum/Guideline

Busan 77 administered orally at 0, 2000, 6000 and 18,000 ppm 

(0, 100, 300 and 900 mg/kg./day)

50 rats/sex/dose

Chronic:

NOAEL = 100 mg/kg/day

LOAEL = 300 mg/kg/day, based on reduced body weight gain (F), 

reduced albumin (M), reduced total protein (M), reduced phosphorus 

(inorganic) (M&F), increased urine pH (F) and a dose related-related 

increase in bloody crusts.

Carcinogenicity:

In females, possible increased incidence of thyroid C-cell adenoma at 

300 and 900 mg/kg/day, and positive trend for this tumor in both sexes.
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

The RfD Committee considered the data in this study and classified 

Busan 77as a Group “D” carcinogen.

Mutagenicity

870.5100

Reverse mutation assay – Salmonella 

typhimurium.

Purity: not reported

MRID 41573701

Lawler, T.L. and DaCosta, K. (1990). Mutagenicity Test on 

Busan 77 in the SalmonellaJMammalian—Microsome Reverse 

Mutation Assay with Confirmatory Assay. Buckman 

Laboratories, Inc. Study #: 12144-0-401R.

Acceptable

Busan 77 doses of 66.7, 100, 333, 667, 1000 and 3330μg/plate

Strains: salmonella (TA1535, TA1537, TA1538, TA98 and 

TA100

Negative

In the initial mutagenic assay (Table 3), using strains TA1535,

TA1537, TA1538, TA100 and TA98 in the presence and absence of S9 

activating system, at doses of 66.7, 100, 333, 667, 1,000 and 3,330 

Mg/plate, did not increase in the number of histidine revertants were 

observed, either in the presence or absence of S9 system. The 

experiment was repeated and all data were acceptable except for strain 

TA9B for which the means were outside the acceptable range both in 

the presence and absence of S9. The TA98 strain was retested both in 

the presence/absence of activation system and the results were 

acceptable (Table 4). Revertants frequencies for all doses of Busan 77 

in all strains approximated or were less than those observed in the 

concurrent negative controls (Tables 1 and 2). All positive controls 

produced positive responses expected of the chemicals.

Based on the results of the Salmonella Reverse Mutation Assay the test 

article Busan 77 did not cause a positive increase in the number of 

histidine revertants per plate.
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

870.5275

Sex-linked recessive lethal test

Purity: 100%

MRID 00151205

McCarroll, N. (1985). Drosophila Sex-linked Recessive Lethal 

Assay [with WSCP (Busan 77)]: Hazelton Laboratories, Inc. 

Project #: 197-183. Unpublished

Acceptable 

Busan 77 administered at 0.08, 0.3 and 0.8 mg/ml

Test strain: drosophila melanogaster

Negative

A dose-related increase in sterility was observed in the mated P1 males 

to increasing concentration of the test material when compared to the 

solvent control. No significant difference in the percentage of sexlinked recessive lethals was observed between the Busan-77 treated 

groups and the medium control group.

870.5385

In vivo micronucleus assay (mice)

Purity: 100%

MRID 00151206/93062021

Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of 

MRID 00151206. Busan 77- in Vivo Mouse Micronucleus 

Assay: Laboratory ID 197-182. Prepared by Hazleton 

Biotechnologies Corporation. 8 p.

Acceptable

Busan 77 administered via gavage at 0.08, 0.3 and 0.8 mg/ml

15 mice/sex/dose

Negative

The test compound WSCP (Bu5an 77) is considered non-mutagenic in 

mice under the condition of this assay.

The test compound WSCP (Busan 77) did not induce a significant 

increase of the PCE containing micronuclei in the treated mice when 

compared to that of the vehicle control value.

The positive control compound cyclophosphamide induced a 

significant increase of the PCE containing micronuclei, which 

indicated the sensitivity of the assay system

(p < 0.0001).

870.5550

Unscheduled DNA synthesis in 

mammalian cells in culture (rat)

Purity: not reported

MRID 40978701/93062022

Cifone, M. (1989) Mutagenicity Test on Busan 77 in the in 

vivo/in vitro Rat Primary Hepatocyte Unscheduled DNA 

Synthesis Assay: Project ID: HLA Study No. 10280-0-494. 

Unpublished study pre- pared by Hazleton Laboratories 

America, Inc. 26 p.

Acceptable

Busan 77 administered at 188, 375, 750 and 1500 mg/kg 

3 rats/group 

Negative

Hepatocyte viability/attachment efficiency ranged from 85.4 to 

95.2%/66.2 to 81.5%. Busan at a concentration of 188, 375, 750 and 

1,500 mg/kg resulted in hepatocyte viability (attached cells) of 93.9, 

93.2, 94.9 and 93.3%, respectively; i.e., no toxicity (see Appendix 2).

None of the treatments with Busan resulted in a significant increase in 

nuclear labeling, when compared to the controls and did not meet the 

minimum criteria for UDS assay for mean net nuclear grain count 

exceeding 6, or at least 10% of the nuclei containing 6 or more grains, 

or at least 2% of the nuclei containing 20 or more grains, to conclude a 

positive response (see Appendix 1). Furthermore, there was no doserelated trend observed. Positive control treated cultures exhibited 

greatly exceeded all three criteria used to indicate UDS. 
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Guideline Number/

Study Type

Test Substance (% a.i.)

MRID Number (Year) /Citation/

Classification/ Doses

Results

870.7485

Metabolism and pharmacokinetics 

(rat)

Purity: 60%

MRID 40268601/93062024

Puhl, R. (1987) Metabolism Study with Busan 77 (WSCP) in 

Rats: Final Report: Laboratory Project ID: HLA 6176-115. 

Unpublished study prepared by Hazleton Laboratories 

America, Inc. 98 p.

Acceptable/Guideline

14C-labeled Busan 77 administered oral and i.v. single of 10 

mg/kg; single oral dose of 1000 mg/kg; repeated oral dose of 

10 mg/kg

5 rats/sex/dose

Intravenous excretion at 10 mg/kg; urine, 38-44% feces, 11-14%.

Tissue contained 43-55% of the dose. Single and repeated oral dose 

excretion: urine, 3% of dose, feces 85-105% of dose. Tissue levels low 

in these groups (0.14% highest levels in any tissue)

Oral 1000 mg/kg dose: urine, 14-17%; feces 68-71% of dose.

Expired CO2 was not detected. In the intravenous dose group, the 

major routes of excretion of radioactivity were via urine and feces. 

Over a 7-day period, approximately 52-55% of the test material 

administered was excreted in the urine (38—44%) and feces (11—

14%); 43—55% was, found in the tissues. In a single repeated oral 

dosed groups 88-106% of the test compound administered was 

excreted in the urine (3%) and feces (85—105%); kidneys, liver, and 

spleen had a highest residue level. In these groups, tissue residue levels 

were low in all tissues, except kidneys. Not more than 0.14% of the 

administered dose was recovered in tissues. Potential for 

bioaccumulation at the single, or repeated oral is minimal. At the 

highest oral dose, 85% of the administered dose was recovered in urine 

(14—17%) and feces (68—71%). The information provided by the 

sponsor explains that almost the entire administered dose was excreted 

in the feces unchanged and remaining 40% of the composition of the 

technical or radiolabeled material was water.

870.7600

Dermal penetration (rat)

Purity: 60%

MRID 40139201/93062025

Bosch, A. (1987) Dermal Absorption of [Carbon 14]-Busan 77 

(WSCP) in Male Rats: HLA Study No. 6176-117: Final 

Report. Unpublished study prepared by Hazleton Laboratories 

America, Inc. 52 p.

Acceptable/Guideline

single dermal administration to male rats at 2, 20, or 200 

mg/rat (0.16, 1.59, or 15.97 mg/cm2, respectively)

24 rats/group

The total amount of Busan 77 absorbed (expressed as percent of 14Cdose) from rats at various times following dermal administration of 

‘4C-Busan 77 at 2, 20 or 200 mg/animal is negligible (less than 0.2% 

of the dose). Also, no radioactivity was found in carcass and blood of 

all treated animals indicating no dermal absorption of Busan 77.

Only less than 0.2% of the Busan 77 was absorbed. Mean % recovery 

of radioactive dose ranged from 74.5 - 92.7%. Majority of the dose was 

recovered in the skin rinse (65.5% - 88.6%) and at the skin site (0.4% -

13.7%). 
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Toxicology References

MRID 00151205 McCarroll, N. (1985) Drosophila Sex-linked Recessive Lethal Assay [with 

WSCP (Busan 77)]: Final Report: Project No. 197-183. Un- published study prepared by 

Hazleton Laboratories. 20 p.

MRID 00151206 Cortina, T. (1984) In vivo Micronucleus Assay in Mice: WSCP: Final Report: 

Project No. 197-182. Unpublished study prepared by Hazleton Laboratories America, Inc. 

38 p.

MRID 40025001 Tisdel, M. (1986) Busan 77--Thirteen-week Subchronic Toxicity Study in 

Rats: Project ID: Study No. 6176-106. Unpublished study prepared by Hazleton 

Laboratories of America, Inc. 285 p.

MRID 40139201 Bosch, A. (1987) Dermal Absorption of [Carbon 14]-Busan 77 (WSCP) in 

Male Rats: HLA Study No. 6176-117: Final Report. Unpublished study prepared by 

Hazleton Laboratories America, Inc. 52 p.

MRID 40170601 Spindler, M. (1987) 13-Week Dermal Toxicity Study with Busan 77 in 

Rabbits: Laboratory Project ID: HLA 6176-118. Unpublished study prepared by Hazleton 

Laboratories America, Inc. 290 p.

MRID 40268601 Puhl, R. (1987) Metabolism Study with Busan 77 (WSCP) in Rats: Final 

Report: Laboratory Project ID: HLA 6176-115. Unpublished study prepared by Hazleton 

Laboratories America, Inc. 98 p.

MRID 40578201 MacKenzie, K. (1988) Two-Generation Reproduction Study with Busan 77 in 

Rats (One Litter per Generation): Project ID: HLA 6176-104. Unpublished study prepared 

by Hazleton Laboratories America, Inc. 952 p.

MRID 40750301 Glaza, S. (1987) Dermal Sensitization Study in Guinea Pigs with WSCP: Proj. 

ID 70303995. Unpublished study prepared by Hazleton Laboratories America, Inc. 30 p.

MRID 40978701 Cifone, M. (1989) Mutagenicity Test on Busan 77 in the in vivo/in vitro Rat 

Primary Hepatocyte Unscheduled DNA Synthesis Assay: Project ID: HLA Study No. 

10280-0-494. Unpublished study pre- pared by Hazleton Laboratories America, Inc. 26 p.

MRID 41234501 Kehoe, D. (1989) One-Year Chronic Toxicity Study with Busan 77 in Dogs: 

Laboratory Project ID HLA 6176-111. Unpublished study prepared by Hazleton 

Laboratories America, Inc. 301 p.

MRID 41248001 Rodwell, D. (1989) Teratology Study in rabbits with Busan 77: SLS Study No. 

3138.29. Unpublished study prepared by Springborn Life Sciences, Inc. 229 p.

MRID 41298601 Rush, R. (1989) Primary Skin Irritation Study in Rabbits with Busan 77: SLS 

Study No. 3138.49. Unpublished study prepared by Springborn Laboratories, Inc. 19 p.
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MRID 41327101 Rush, R. (1989) Acute Dermal Toxicity Study in Rabbits with Busan 77: Lab 

Project Number: 3138.50. Unpublished study prepared by Springborn Laboratories, Inc. 26 

p.

MRID 41361701 Rush, R. (1989) Primary Eye Irritation Study in Rabbits with Busan 77 (EPAFIFRA): Lab Project Number: 3138/52. Unpublished study prepared by Springborn 

Laboratories, Inc. 25 p.

MRID 41373401 Rush, R. (1990) Acute Oral Toxicity Study in Rats with Busan 77: SLS Study 

No. 3138.51. Unpublished study prepared by Springborn Laboratories, Inc. 65 p.

MRID 41423001 Nemec, M. (1987) A Teratology Study in Rats with Busan 77: Final Report: 

Lab Project Number: WIL-94020. Unpublished study prepared by WIL Research 

Laboratories, Inc. 281 p.

MRID 41494301 Kehoe, D. (1990) 18-Month Study with Busan 77 in Mice: Lab Project 

Number: HLA 6176-109. Unpublished study prepared by Hazleton Labs America, Inc. 924 

p.

MRID 41561301 Kehoe, D. (1990) Combined Chronic Toxicity and Carcinogenicity Study with 

Busan 77 in Rats: Draft Report: Lab Project Number: HLA 6176-107. Unpublished study 

prepared by Hazleton Laboratories America, Inc. 1511 p.

MRID 41573701 Lawlor, T.; DaCosta, K. (1990) Mutagenicity Test on Busan 77 in the 

Salmonella/Mammalian-Microsome Reverse Mutation Assay (Ames Test) with 

Confirmatory Assay: Lab Project Number: 12144-0-401R. Unpublished study prepared by 

Hazleton Laboratories America, Inc. 33 p.

MRID 41809101 Kehoe, D. (1991) Combined Chronic Toxicity and Carcinogenicity Study with 

Busan 77 in Rats: Final Report (Supplement to MRID 41561301): Lab Project Number 

6176-107. Unpublished study pre- pared by Hazleton Labs America, Inc. 1511 p.

MRID 41877501 Hoffman, G. (1991) An Acute Inhalation Toxicity Study of Busan 77 in the 

Rat: Lab Project Number: 90-8305. Unpublished study prepared by Bio/dynamics, Inc. 105 

p.

MRID 93062009 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 41373401. 

Busan 77- Acute Oral Toxicity Study in Rats: Laboratory ID: 3138/51. Prepared by 

Springborn Laboratories, Inc. 9 p.

MRID 93062010 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 41327101. 

Busan 77- Acute Dermal Toxicity Study in Rabbits: Laboratory ID 3138/50. Prepared by 

Springborn Laboratories, Inc. 6 p.
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MRID 93062011 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 41361701. 

Busan 77- Primary Eye Irritation in the Rabbit: Laboratory ID 3138.52. Prepared by 

Springborn Laboratories, Inc. 7 p.

MRID 93062012 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 41298601. 

Busan 77- Primary Dermal Irritation in the Rabbit: Laboratory ID 3138/49. Prepared by 

Springborn Laboratories, Inc. 6 p.

MRID 93062013 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 40750301. 

Busan 77- Dermal Sensitization in the Guinea Pig (Close Patch Technique): Laboratory ID 

70303995. Prepared by Hazleton Laboratories America, Inc. 6 p.

MRID 93062014 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 40025001. 

Busan 77- Thirteen Week Subchronic Toxicity Study in Rats Laboratory ID: HLA 6176-

106. Prepared by Hazleton Laboratories America, Inc. 12 p.

MRID 93062015 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 40170601. 

Busan 77- 13 Week Dermal Toxicity Study in Rabbits: Laboratory ID 6176-118. Prepared 

by Hazleton Laboratories America, Inc. 6 p.

MRID 93062016 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 41234501. 

Busan 77- One Year Chronic Toxicity Study in Dogs: Laboratory ID HLA 6176-111. 

Prepared by Hazleton Laboratories America, Inc. 8 p.

MRID 93062017 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 41494301. 

Busan 77- 18 Month Study in Mice: Laboratory ID: HLA 6176-109. Prepared by Hazleton 

Laboratories America, Inc. 10 p.

MRID 93062019 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 41248001. 

Busan 77- Teratology Study in Rabbits: Laboratory ID: SLS Study No. 3138-29. Prepared 

by Springborn Laboratories, Inc. 9 p.

MRID 93062020 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 40578201. 

Busan 77- Two Generation Reproduction Study in Rats: Laboratory ID HLA 6176-104. 

Prepared by Hazleton Laboratories America, Inc. 9 p.

MRID 93062021 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 00151206. 

Busan 77- in Vivo Mouse Micronucleus Assay: Laboratory ID 197-182. Prepared by 

Hazleton Biotechnologies Corporation. 8 p.

MRID 93062022 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 40978701. 

Busan 77- in Vivo/In Vitro Rat Primary Hepatocyte Unscheduled DNA Synthesis Assay: 

Laboratory ID: 10280-0-494. Prepared by Hazleton Laboratories America, Inc. 8 p.
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MRID 93062024 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 40268601.

Busan 77- Metabolism Study in Rats: Laboratory ID HLA 6176-115. Prepared by Hazleton 

Laboratories America, Inc. 17 p.

MRID 93062025 Drake, K. (1990) Buckman Labs Inc. Phase 3 Summary of MRID 40139201. 

Busan 77- Dermal Absorption: Laboratory ID: HLA 6176-117. Prepared by Hazleton 

Laboratories America, Inc. 11 p.

EPA DOCUMENTS

EPA (2007a) Poly(oxyethylene(dimethylminino) ethylene(dimethyliminio) ethylenedichloride) 

Busan 77 – Revised Report of the Antimicrobials Division’s Toxicology Endpoint Selection 

Committee (ADTC), 7/10/07, Timothy F. McMahon, Ph.D.

EPA (2007b) Poly(oxyethylene(dimethylminino) ethylene(dimethyliminio) ethylenedichloride) 

Busan 77: Toxicology Disciplinary Chapter for the Reregistration Eligibility Decision 

Document, PC Code 069183, Case 3034, Antimicrobials Division, 7/26/07, Timothy F. 

McMahon, Ph.D.

EPA (2007c) Poly(oxyethylene(dimethylminino)ethylene(dimethyliminio) ethylenedichloride): 

Preliminary Risk Assessment for the Reregistration Eligibility Decision, PC Code 069183, 

Case 3034, Antimicrobials Division, 8/04/07. 
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Appendix B Environmental Fate

Environmental Fate and Transport Properties of Busan 77

Busan 77 is miscible in water (Table 4) and is not expected to degrade by either abiotic processes 

(MRIDs 41407401 and 41420901) or biotic processes (MRIDs 40334101, 40165201, 40165202).

Movement from water or soil into air is also not expected to occur most uses based on the lack of 

vapor pressure and Henry’s Law values (Table 4), but the air washer use has the potential to 

create aerial exposure because the Busan 77 dissolved in water can volatilize with the water.

Bioconcentration in aquatic organisms is also not expected based on the water miscibility and 

low log P values (Table 4). Sorption to soil, sediment, and sludge is expected to be the primary 

route of dissipation based on the fact that is a quaternary ammonium compound that has a 

positive electrical charge. However, data for sorption to soil and sludge have not been submitted 

and are anticipated to be required. Busan 77 has the potential to reach WWTPs from the 

registered uses, and data on activated sludge sorption isotherm (ASSI) and activated sludge 

respiration inhibition (ASRI). WWTP biodegradability studies (835.3110, 835.3220, 835.3240, 

and 835.3280) are normally required for chemicals that, based on the registered uses, have the 

potential to reach wastewater treatment plants. However, in the case of Busan 77, there is no 

apparent route of chemical or microbial degradation, and therefore degradation data in a WWTP 

are not anticipated to be required. 

Water and Sediment

Hydrolysis and Aqueous Photolysis

Busan 77 is stable to abiotic hydrolysis and to photodegradation in water at pH 5, 7, and 9 at 25 o

C (MRID 41407401 for hydrolysis and MRID 41420901 for photodegradation in water).

Octanol-Water Partition Coefficient and Bioconcentration in Fish

Busan 77 is miscible in water (Table 4), and as a result, is not expected to bioconcentrate in fish 

and other aquatic organisms.

Aerobic Aquatic Metabolism

Busan 77 was essentially stable under aerobic aquatic conditions. Sediment percentages of 

applied material increased from 30.5 % at time zero to 93.5-106.6 % by 7-31 days after 

treatment, and water levels decreased from 66.1 % at time zero to 1.2-5.2 % by 7-31 days

(MRID 40334101).
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Anaerobic Aquatic Metabolism

Busan 77 was essentially stable under anaerobic aquatic conditions. Sediment percentages of 

applied material were approximately 100 % of the total, and water never contained more than 2.2 

% of applied (MRID 40165201).

Soil

Soil Leaching/Adsorption/Desorption Batch Equilibrium

Batch equilibrium studies on Busan 77 have not been submitted and are anticipated to be 

required.

Aerobic Soil Metabolism

Based on a cursory review, Busan 77 was stable in aerobic soil at 23-25 oC (MRID 40165202).

This study will be evaluated in Registration Review.

Anaerobic Soil Metabolism

Data on anaerobic soil metabolism were not submitted, but they are not anticipated to be required 

because acceptable data were submitted for anaerobic aquatic metabolism.

Fate and Transport in WWTP

Activated Sludge Sorption Isotherm

Data on activated sludge sorption isotherm (ASSI) sorption to activated sludge are anticipated to 

be required because the registered uses of Busan 77 can create exposure to wastewater treatment 

plants (WWTPs). While the 158W regulations do not require ASSI data for those compounds 

with log P (log Kow) values of < 3, Busan 77 is a quaternary ammonium compound that is 

expected to sorb to sludge because of its positive electrical charge.

Activated Sludge Respiration Inhibition

Data on activated sludge respiration inhibition (ASRI) are anticipated to be required because the 

registered uses of Busan 77 can create exposure to WWTPs.

Activated Sludge Biodegradation

Busan 77 has the potential to reach WWTPs from the registered uses, and data on activated 

sludge sorption isotherm (ASSI) and activated sludge respiration inhibition (ASRI) are 

anticipated to be required. WWTP biodegradability studies (835.3110, 835.3220, 835.3240, and 

835.3280) are normally required for chemicals that, based on the registered uses, have the 

potential to reach wastewater treatment plants. However, in the case of Busan 77, there is no 

apparent route of chemical or microbial degradation, and therefore degradation data in a WWTP 

are not anticipated to be required.
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Environmental Fate References

MRID 40165201 Obrist, J. (1987) Anaerobic Aquatic Metabolism of Busan 77: Final Report: 

Laboratory Project Identification: HLA 6015-250. Unpublished study prepared by Hazleton 

Laboratories America, Inc. 50 p.

MRID 40165202 Das, Y. (1987) Aerobic Soil Metabolism of WSCP: Biospheric No. 84E-301A. 

Unpublished study prepared by Biospheric Inc. 50 p.

MRID 40334101 Obrist, J. (1987) Aerobic Aquatic Metabolism of Busan 77: Laboratory Project 

ID: HLA 6015-251: Final Report. Unpublished study prepared by Hazleton Laboratories 

America, Inc. 69 p.

MRID 41407401 Smith, A. (1990) Determination of Aqueous Hydrolysis Rate Constant and 

Half-life of Busan 77: Final Report: Lab Project Number: 995-1089-6116-715: Lab Report 

Number 90-2-3216. Unpublished study prepared by Springborn Laboratories, Inc. 32 p.

MRID 41420901 Smith, A. (1990) Determination of Aqueous Photolysis Rate Constant and 

Half-life of Busan 77: Lab Project Number: 995-1089-6117-720; 90-3-3251. Unpublished study 

prepared by Springborn Laboratories, Inc. 37 p.
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Appendix C Ecotoxicology Profile

Toxicity to Terrestrial Receptors

Birds

In order to establish the toxicity of Busan 77 to avian species, the Agency requires an acute oral 

toxicity study using the technical grade active ingredient (TGAI). The preferred-test species is 

either mallard duck (a waterfowl) or bobwhite quail (an upland game bird). The results of two 

acute oral toxicity studies, submitted for Busan 77, are provided in the following table (Table 

C1).

Table C1 – Acute Oral Toxicity of Busan 77 to Birds

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(mg/kg a.i.)

Toxicity 

Category

Satisfies 

Guidelines/

Comments

Reference

(MRID No.

unless 

specified)

Mallard duck

(Anas 

platyrhynchos)

Busan 77 

61.7%

LD50 = 497

NOAEL = 51

Moderately 

toxic

Yes (core)

- 14-day test 

duration

- 36 weeks of age

41654801

Bobwhite quail

(Colinus 

virginianus)

Busan 77 

% purity 

unknown

LD50 = 690

NOAEL = <172

(Based on the 

assumption of 

60% a.i.)

Slightly 

toxic

No (supplemental)

- 21-day test 

duration

- young adult

- study conducted 

before adoption of 

GLP principles

ID 0522-010-

20

These two acceptable acute oral toxicity studies indicate that Busan 77 is moderately toxic to 

slightly toxic to birds on an acute oral basis. 

Birds, Subacute

A subacute dietary study using the TGAI may be required on a case-by-case basis depending on 

the results of lower-tier ecological studies and pertinent environmental fate characteristics in 

order to establish the toxicity of a chemical to avian species. This testing was required for Busan 

77. The preferred-test species is either the mallard duck or bobwhite quail. The results of two 
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subacute dietary toxicity studies, submitted for Busan 77, are provided in the following table 

(Table C2).

Table C2 – Subacute Oral Toxicity of Busan 77 to Birds

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(ppm a.i.)

Toxicity 

Category

Satisfies Guidelines/

Comments

Reference

(MRID 

No.)

Mallard duck

(Anas 

platyrhynchos)

Busan 77 

60.3%

LC50 (diet) = 

>12,000

NOAEC = 

3,000

Relatively 

nontoxic

No (supplemental)

- 8-day test duration

- 10 days of age

- study conducted 

before adoption of 

GLP principles

41411501

Bobwhite quail

(Colinus 

virginianus)

Busan 77 

61.6%

LC50 (diet) = 

>3,462

NOAEC = 

3,462

Slightly 

toxic

Yes (core)

- 8-day test duration

- 11 days of age 

00159307

The results of these two acceptable studies indicate that Busan 77 is slightly toxic to relatively 

nontoxic to avian species through subacute dietary exposure. 

Mammals

Acute and Chronic Toxicity

Wild mammal testing is not required by the Agency. In most cases, rat toxicity values obtained 

from studies conducted to support data requirements for human health risk assessments substitute 

for wild mammal testing.

Terrestrial Plants

One study that evaluates the toxicity of Busan 77 to several species of terrestrial plants (seedling 

emergence-tier I and vegetative vigor-tier I and tier II) and five studies that evaluate the toxicity 

of Busan 77 to freshwater aquatic plants have been submitted. Results of the terrestrial plant 

study are presented in Tables C3-C5 and results of the aquatic plant studies are presented in 

Table C10.
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Table C3 – Seedling Emergence (Tier I) Toxicity of Busan 77 to Terrestrial Plants*

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(ppm a.i.)*

Satisfies Guidelines/

Comments

Reference

(MRID No.)

Monocot

Oat (Avena sativa)

Busan 77 

60.3%

EC25 = >73

NOAEC = 73

Yes (core)

- 14-day test duration

- most sensitive 

parameter: shoot 

length (14% 

reduction)

42038101

Dicot

Lettuce (Lactuca 

sativa)

Busan 77 

60.3%

EC25 = >73

NOAEC = 73

Yes (core)

- 14-day test duration

- most sensitive

parameter: shoot 

length (17% reduction)

42038101

*Data are listed for most sensitive species and most sensitive parameter. Species tested were cabbage, carrot, cucumber, lettuce, 

soybean, tomato, corn, oats, onion and ryegrass.

Table C4 – Vegetative Vigor (Tier I) Toxicity of Busan 77 to Terrestrial Plants

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint**

(ppm a.i.)

Satisfies Guidelines/

Comments

Reference

(MRID No.)

Monocot

Corn (Zea mays)

Busan 77 

60.3%

EC25 = >74

NOAEC = 74

Yes (core)

-14-day test duration

-most sensitive 

parameter: shoot 

weight (39% 

reduction)

42038101

Dicot

Cucumber 

(Cucumis sativus)

Busan 77 

60.3%

EC25 = >74

NOAEC = 74

Yes (core)

- 14-day test duration

- most sensitive

parameter: root weight 

(31% reduction)

42038101

*Data are listed for most sensitive species and most sensitive parameter. Species tested were cabbage, carrot, cucumber, lettuce, 

soybean, tomato, corn, oats, onion and ryegrass.

**EC25 value for corn based on shoot length and root weight. EC25 for cucumber based on shoot length and shoot weight.
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Table C5 – Vegetative Vigor (Tier II) To Toxicity of Busan 77 to Terrestrial Plants*

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint*

(ppm a.i.)

Satisfies Guidelines/

Comments

Reference

(MRID No.)

Monocot

Ryegrass (Lolium 

perenne)

Busan 77 

60.3%

EC25 = >74

NOAEC = 74

No (supplemental)

- 14-day test duration

- most sensitive 

parameter: root 

weight (0% 

reduction)

42038101

Dicot

Tomato 

(Lycopersicon 

esculentum)

Busan 77 

60.3%

EC25 = >74

NOAEC = 74

No (supplemental)

- 14-day test duration

- most sensitive

parameter: shoot 

weight (15% 

reduction)

42038101

*Data are listed for most sensitive species and most sensitive parameter.

Toxicity to Aquatic Receptors

Freshwater Fish

In order to establish the acute toxicity of Busan 77 to freshwater fish, the Agency requires

freshwater fish toxicity studies using the TGAI. The preferred test species are rainbow trout (a 

cold water fish) and bluegill sunfish (a warm water fish). The results of 3 freshwater fish acute 

studies submitted for Busan are presented in Table C6.

Table C6 – Acute Toxicity of Busan 77 to Freshwater Fish

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(mg a.i./L)

Toxicity 

Category

Satisfies Guidelines/

Comments

Reference

(MRID 

No.)

Rainbow Trout 

(Oncorhynchus 

mykiss)

Busan 77 

61.9%

LC50 = 0.047

NOAEC = 

0.037

Very 

highly 

toxic

Yes (core)

- 96-hr test duration

- static test system

41352001
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Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(mg a.i./L)

Toxicity 

Category

Satisfies Guidelines/

Comments

Reference

(MRID 

No.)

Bluegill sunfish 

(Lepomis 

macrochirus)

Busan 77 

61.9%

LC50 = 0.21 

NOAEC = 

0.13

Highly 

toxic

Yes (core)

- 96-hr test duration

- static test system

41352002

Rainbow Trout 

(Oncorhynchus 

mykiss)

Busan 77 

60%

LC50 = 0.26

NOAEC = 

0.11

Highly 

toxic

No (supplemental)

- 96-hr test duration

- static test system

- study lacks 

important 

information

- study conducted 

before adoption of 

GLP principles

00107207

Freshwater acute toxicity tests indicate that Busan 77 is very highly toxic to highly toxic to fish 

on an acute basis.

Freshwater Invertebrates

The Agency requires a freshwater aquatic invertebrate study using the TGAI to establish the 

acute toxicity to freshwater invertebrates. The preferred test species is Daphnia magna. The 

results of two studies submitted for Busan 77 are provided in the following table (Table C7).

Table C7 – Acute Toxicity of Busan 77 to Freshwater Invertebrates

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(mg a.i./L)

Toxicity 

Category

Satisfies 

Guidelines/

Comments

Reference

(MRID 

No.)

Waterflea 

(Daphnia 

magna)

Busan 77

61.9%

EC50 = 0.280 

NOAEC = 

0.130

Highly 

toxic

Yes (core)

- 48-hr test 

duration

- static test 

system

41352003
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Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(mg a.i./L)

Toxicity 

Category

Satisfies 

Guidelines/

Comments

Reference

(MRID 

No.)

Chironomus

riparius

Busan 77

99.8%

LC50 = 3.56 in 

overlying 

water

LC50 = >966 in 

sediment

Moderately 

toxic in 

water

Practically 

nontoxic in 

sediment

Yes 

(supplemental)

-28-day test 

duration

-insufficient 

number of 

replications

48715801

The results of the waterflea study indicates that Busan 77 is highly toxic to freshwater 

invertebrates on an acute basis. The sediment study indicated that Busan 77 is moderately toxic 

to freshwater invertebrates in the overlying water and practically nontoxic in the sediment. 

Estuarine/Marine Fish and Invertebrates

Acute toxicity testing with estuarine and marine organisms using the TGAI is required when the 

end-use product is intended for direct application to the marine/estuarine environment or effluent 

containing the active ingredient is expected to reach this environment. The preferred fish test 

species is the sheepshead minnow. The preferred invertebrate test species are mysid shrimp and 

eastern oysters. This testing is required for Busan 77 based on the chemical’s potential to reach 

estuarine and marine environments. The results of three toxicity studies submitted for Busan 77 

are presented in Table C8.

Table C8 – Acute Toxicity of Busan 77 to Estuarine and Marine Organisms

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(mg a.i./L)

Toxicity 

Category

Satisfies Guidelines/

Comments

Reference

(MRID 

No.)

Sheepshead 

minnow 

(Cyprinodon 

variegates)

Busan 77

60.0%

LC50 > 360

NOAEC = 

360

Relatively 

nontoxic

Yes (core)

- 96-hr test duration 

- static test system

40139202
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Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(mg a.i./L)

Toxicity 

Category

Satisfies Guidelines/

Comments

Reference

(MRID 

No.)

Mysid shrimp 

(Mysidopsis 

bahia)

Busan 77

60.0%

LC50 = 7.8

NOAEC <7.8

Moderately 

toxic

Yes (core)

- 96-hr test duration 

- static test system

40139203

Quahog clam 

(Mercenaria 

mercenaria)

Busan 77

60.0%

EC50 = 0.21

NOAEC = 

0.14

Highly 

toxic

Yes (core)

- 48-hr test duration 

- static test system

- embryo/larval stage

40334201

The results of these three acceptable studies indicate that Busan 77 is highly toxic to moderately 

toxic to estuarine/marine invertebrates on an acute basis and relatively nontoxic to 

estuarine/marine fish on an acute basis.

Chronic Toxicity

Chronic toxicity testing (fish early life stage and aquatic invertebrate life cycle) is required for 

pesticides when certain conditions of use and environmental fate apply. The preferred freshwater 

fish test species is the fathead minnow. The preferred freshwater invertebrate is Daphnia magna.

This testing is required for the Busan 77 once-through cooling water uses. The results of two 

toxicity studies submitted for Busan 77 are presented in Table C9.
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Table C9 – Chronic Toxicity of Busan 77 to Freshwater Organisms 

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint

(mg a.i./L)

Satisfies 

Guidelines/

Comments

Reference

(MRID No.)

Waterflea 

(Daphnia magna)

Busan 77

60.3%

LOEC = 0.020

NOAEC = 

0.012

No (supplemental)

- 21-day test

duration 

- static renewal test 

system

- organisms not 

distributed or fed 

properly

- excess mortality

42479601

Fathead minnow 

(Pimephales 

promelas)

Busan 77 

(59.8% ai)

LOAEC 33 ug 

ai/L

NOAEC 19 ug 

ai/L

Yes (Acceptable) 

-flow-through

49453501

The fathead minnow study indicates that Busan 77 is very highly toxic to fathead minnow. The 

study on the waterflea does not fulfill the guideline requirement for a chronic aquatic 

invertebrate study.

Aquatic Plants

Non-target plant phytotoxicity testing is required for pesticides when certain conditions of use 

and environmental fate apply. This testing is required for the once-through cooling water uses of 

Busan 77. Testing has been conducted with Busan 77 on several aquatic plant species. Testing is 

normally conducted with one species of aquatic vascular plant (Lemna gibba) and four species of 

algae: (1) freshwater green alga, Selenastrum capricornutum, (2) marine diatom, Skeletonema 

costatum, (3) freshwater diatom, Navicula pelliculosa, and (4) bluegreen cyanobacteria, 

Anabaena flos-aquae. The rooted aquatic macrophyte rice (Oryza sativa) is also tested in 

seedling emergence and vegetative vigor tests.
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Table C10 – Toxicity of Busan 77 to Aquatic Plants

Species

Chemical,

% Active 

Ingredient

(a.i.)

Tested

Endpoint 

(mg a.i./L)

Satisfies Guidelines/

Comments

Reference

(MRID No.)

Marine diatom 

(Skeletonema 

costatum)

Busan 77

60.3%

EC50 = 0.090

NOEC < 0.024

No (supplemental)

- 120-hour test duration

- static test system

42013302

Freshwater 

diatom 

(Navicula 

pelliculosa)

Busan 77

60.3%

EC50 = 0.083

NOEC = 0.044

Yes (core)

- 96-hour test duration

- static test system

42013303

Bluegreen 

cyanobacteria 

(Anabaena flosaquae)

Busan 77

60.3%

EC50 = 0.11

NOEC = 0.05

Yes (core)

- 120-hour test duration

- static test system

42013304

Green algae

(Selenastrum 

capricornutum)

Busan 77

60.3%

EC50 = 0.0088

NOEC < 0.001

No (supplemental)

- 120-hour test duration

- static test system

42013305

Duckweed 

(Lemna gibba)

Busan 77

60.3%

EC50 > 0.65

NOEC = 0.043

No (supplemental)

-14-day test duration

-static renewal test 

system

- maximum concentration 

tested produced only 30% 

growth inhibition

42013301

Green alga 

(Pseudokirchner

iella 

subcapitata)

Busan 77

59.8%

EC50 = 3.0 ug/L Yes (supplemental)

-96-hour test duration

-test material sorbing to 

test vessel glass

49453501
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Ecotoxicity References

MRID 00107207 Knott, W.B. 1970. Busan 77 (WSCP) - Evaluation of Acute LC50 for Rainbow 

Trout. Study performed by Woodward Research Corporation for Buckman Laboratories, 

Inc., Memphis, TN.

MRID 00159307 Beaver, J.B. 1985. Busan 77 - A Dietary LC50 Study with the Bobwhite.

Project No. 210-104. Study performed by Wildlife International, Ltd. for Buckman

Laboratories, Inc., Memphis, TN.

MRID 052201020 WARF Institute, Inc. 1971. Report on Acute Oral LD50 in Bobwhite Quail.

Prepared for Buckman Laboratories, Inc., Memphis, TN.

MRID 40139202 Surprenant, D.C. 1987a. Acute Toxicity of Busan 77 to Sheepshead Minnow 

(Cyprinodon variegates). Report No. BW-87-2-2294. Study performed by Springborn 

Bionomics, Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 40139203 Surprenant, D.C. 1987b. Acute Toxicity of Busan 77 to Mysid Shrimp 

(Mysidopsis bahia). Report No. BW-87-3-2346. Study performed by Springborn Bionomics, 

Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 40334201 Surprenant, D.C. 1987c. Acute Toxicity of Busan 77 to Embryos-Larvae of the 

Quahog Clam (Mercenaria mercenaria). Report No. BW-87-8-2459. Study performed by

Springborn Bionomics, Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 41352001 LeLievre, M.K. et al. 1990c. (Busan 77) - Acute Toxicity to Rainbow Trout 

(Oncorhynchus mykiss) Under Static Conditions. Report No. 89-12-3182. Study performed 

by Springborn Laboratories, Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 41352002 LeLievre, M.K. et al. 1990a. "(Busan 77) - Acute Toxicity to Bluegill 

(Lepomis macrochirus) Under Static Conditions. Report No. 89-12-3176. Study performed 

by Springborn Laboratories, Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 41352003 LeLievre, M.K. et al. 1990b. (Busan 77) - Acute Toxicity to Daphnids 

(Daphnia magna) Under Static Conditions. Report No. 89-11-3181. Study performed by 

Springborn Laboratories, Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 41411501 Bodden, R.M. 1974. Busan 77 - Acute Avian Dietary Toxicity Study in 

Mallard Ducks. Report No. 4043621. Study performed by the WARF Institute, Inc. for 

Buckman Laboratories, Inc., Memphis, TN.

MRID 41654801 Campbell, S. et al. 1990. Busan 77: An Acute Oral Toxicity Study with the 

Mallard. Project No. 210-114A. Study performed by Wildlife International, Ltd. for 

Buckman Laboratories, Inc., Memphis, TN.

MRID 42013301 Hoberg, J.R. 1991b. Busan 77 - Toxicity to the Duckweed Lemna gibba G3.

Report No. 91-5-3749. Study performed by Springborn Laboratories, Inc. for Buckman 

Laboratories, Inc., Memphis, TN. 
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MRID 42013302 Hoberg, J.R. 1991c. Busan 77 - Toxicity to the Marine Diatom Skeletonema 

costatum. Report No. 91-9-3864. Study performed by Springborn Laboratories, Inc. for 

Buckman Laboratories, Inc., Memphis, TN.

MRID 42013303 Hoberg, J.R. 1991d. Busan 77 - Toxicity to the Freshwater Diatom Navicula 

pelliculosa. Report No. 91-4-3728. Study performed by Springborn Laboratories, Inc. for 

Buckman Laboratories, Inc., Memphis, TN.

MRID 42013304 Hoberg, J.R. et al. 1991e. Busan 77 - Toxicity to the Freshwater Bluegreen 

Algae Anabaena flos-aquae. Report No. 91-4-3726. Study performed by Springborn 

Laboratories, Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 42013305 Hoberg, J.R. 1991f. Busan 77 - Toxicity to the Freshwater Green Algae 

Selenastrum capricornutum. Report No. 91-4-3732. Study performed by Springborn 

Laboratories, Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 42038101 Hoberg, J.R. 1991a. Busan 77 - Determination of Effects on Seed Germination, 

Seedling Emergence, and Vegetative Vigor of Ten Plant Species. Report No. 91-3-3695.

Study performed by Springborn Laboratories, Inc. for Buckman Laboratories, Inc., 

Memphis, TN.

MRID 42479601 Putt, A.E. et al. 1992. Busan 77 – Chronic Toxicity to Daphnids (Daphnia 

magna) Under Static Renewal Conditions. Report No. 92-7-4343. Study performed by 

Springborn Laboratories, Inc. for Buckman Laboratories, Inc., Memphis, TN.

MRID 48715801 Kranzfelder, J.A., L. Stuerman, and D. Malorin. 1998. Sediment bioassay 

procedure for determining the toxicity of WSCP to Chironomus sp. Report No. 44908. 

Study performed by ABC Laboratories for Buckman Laboratories International, Memphis, 

TN. 

MRID 49362401 Schwader, A.L. 2014. WSCP; Early life-stage toxicity test with fathead 

minnow (Pimephales promelas), following OECD guideline #210 and OSCPP guideline 

850.1400. Smithers Viscient, Wareham, MA. Study No. 995.6187. Submitted by Buckman 

Laboratories, Inc., Memphis, TN.

MRID 49453501 Softcheck, K.A. 2014. [14C]WSCP – 96-hour toxicity test with the freshwater 

green algae, Pseudokirchneriella subcapitata, following OCSPP guideline 850.4500. 

Smithers Viscient, Study No. 995.6190. Sponsored by Buckman Laboratories, Inc., 

Memphis, TN.
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Appendix D Screening Level Down-theDrain Analysis

The Down-the-Drain (DtD) module was not run for Busan 77.

Generally, the Agency uses the DtD module of E-FAST (Exposure and Fate Assessment 

Screening Tool), [version 2] to screen for the potential for aquatic organisms downstream of 

domestic wastewater treatment plants to be exposed to antimicrobial pesticides. 

The Down-the-Drain module uses daily per capita release of household wastewaters, stream 

dilution factors, and wastewater treatment plant removal efficiency to provide both high-end and 

median (average) time-averaged surface water concentrations of chemicals discharged by 

domestic wastewater treatment plants. The high-end scenario uses surface water concentrations 

downstream of domestic wastewater treatment plants based on the 10th percentile stream dilution 

factor (SDF); the average scenario uses surface water concentrations based on the 50th percentile 

SDF. SDF is defined as the ratio of the stream flow downstream of a wastewater treatment plant 

to the wastewater treatment plant flow.

Inputs used to run the DtD module include concentrations of concern (COCs) for aquatic 

organisms, percent removal during wastewater treatment, and hypothetical wastewater treatment 

plant influent volumes. The results of the DtD module are expressed as number of days of 

exceedance of COCs for aquatic organisms. COCs are grouped into 3 categories: acute, 

endangered/listed, and chronic. Acute COCs for freshwater fish and invertebrates are derived by 

dividing LC50 values for freshwater organisms by 2. Endangered/listed COCs for freshwater fish 

and invertebrates are derived by dividing LC50 values by 20. EC50 values for plants are used to 

establish acute COCs for plants. NOAEC values for plants are used to establish 

endangered/listed COCs for plants. Chronic COCs for freshwater fish and invertebrates are based 

on NOAEC values. For acute and endangered listed COCs, there may be potential concern for 

freshwater organisms if the COC is exceeded for 1 day or more. For chronic COCs, there may be 

potential concern for freshwater organisms if the COC is exceeded 20 days or more.

The Down-the-Drain module are run using both a high-end scenario and an average scenario to 

determine whether the model predicted exceedences of concentrations of concern.
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Appendix E Product Chemistry

Table E1 presents the active ingredient to be assessed in case 3034: Busan 77 (PC Code 069183). 

Table E1 – Chemical Identification of Busan 77

Chemical Name Busan 77

Chemical

Classification Polymeric quaternary ammonium 

Chemical Name Poly[oxy-1,2-ethanediyl(dimethyliminio)-1,2-ethanediyl(dimethyliminio)-1,2-

ethanediylchloride (1:2)]

PC Code 069183

CAS Number

31512-74-0

Molecular Formula C10H25N2O (monomer without chlorines)

Molecular Weight 

(grams/mole)

189.32 (monomer)10

3,368 (polymer)11

Molecular Structure

Smiles Code CCOCCN(C)(C)CCN(C)(C) (includes 2 chlorines as counter ions)

The Busan 77 product chemistry information relevant to risk assessment is summarized in Table 

E2.

Table E2 – Physical-Chemical and Environmental Fate Properties for Busan 77

Guideline No./

Study Type

Results Reference/Comments (MRID unless specified)

Water 

Solubility(mg/L)

10,000 Assumed based on miscibility

830.7050

UV visible sorption

N/A 49176301

Compound has no double bonds to absorb UV light

830.7370

Dissociation constant

Waived 47120304/42372401

830.7550

Partition coefficient

Waived 47120304

830.7950

Vapor pressure

Negligible 47120304

Manufacturing Use Product is 60 % polymer and 40 % 

water

Henry’s Law 

Constant

8.5 x 10-12 Volatility from water not likely to occur given the high 

MW, solubility, and positive charges that will enhance 

sorption to solids. 

 10 EPIWEB 4.11

11 Average, 7/10/91 Buckman letter 
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Product Chemistry References

MRID 42372401 Bowles, D. (1991) Product Chemistry for WSCP: Lab Project Number: NA 

909218/1. Unpublished study prepared by Twin City Testing Corp. and NATEC Institut 

fur Naturwissenschaftlich-technische Dienste GmbH. 88p.

MRID 46915602 Wo, C. (2006) Physical and Chemical Characteristics (830 Series, Group B): 

Algae Clean Out W. Unpublished study prepared by Product Safety Laboratories. 6 p.

MRID 47120304 Tillman, A. (2007) Waiver Request for Certain Data Requirements for WSCP 

MUP. Project Number: KT/200704. Unpublished study prepared by Kerley Trading, Inc. 

7p.

MRID 49176301 Dotse, A.; Grabowicz, J. (2013) Product Chemistry for WSKT: Active 

Ingredient and Physical/ Chemical Properties: Final Report. Unpublished study prepared 

by Buckman Laboratories, Inc. 27p.
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Appendix F Comments Received 

Concerning the Preliminary Work Plan

On September 25, 2015 EPA opened a 60-day public comment period on the preliminary work 

plan (PWP) for the registration review of Busan 77. The comment period ended on December 

01, 2015. The comments received did not result in a modification to the anticipated data needs 

detailed in the Busan 77 PWP.

The Agency received submissions from the Physicians Committee for Responsible Medicine and 

Kathleen A. Rose during the public comment period on the initial docket.

Comment: The Physicians Committee for Responsible Medicine recommends that registrant(s) 

apply for waiver for immunotoxicity data using the Agency’s retrospective analysis document 

located at http://www2.epa.gov/pesticide-registration/retrospective-analysis-immunotoxicitystudy.

Agency Response: The Agency thanks the submitter for the information and the web link. The 

Agency reviews all pertinent information when considering waiver requests.

Comment: Ms. Rose is concerned that the use of neonics (neonicotinoids) pose potential risk to 

bees and other pollinators and recommends that the Agency ban neonics. 

Agency Response: The Agency thanks the submitter for their comments. Busan 77 is a 

polymeric quaternary ammonium chemical and not associated with neonicotinoids. The Agency 

provides pollinator reference information at http://www.epa.gov/pollinator-protection.