Docket Number EPA

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P age 2 of 74 HHT Registration Review Team

Human Health & Environmental Effects David Bays Najm Shamim Pat Jennings Nate Mottl Jenny Tao Kristin Rury Don Wilbur

Risk Management Stephen Savage T ina Pham Rose Kyprianou Elizabeth Watkins Seiichi Murasaki

Office of General Counsel Phil ip RossDocket Number EPA

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P age 3 of 74 TERMS, ABBREVIATIONS AND SYMBOLS

A.I. or a.i. active ingredient ASRI a ctivated sludge r espiration i nhibition
atm

- m 3 /mole a tmospheric pressure

- cubic meter per mole °C d egrees Celsius CAS Chemical Abstracts Service CFR Code of Federal Regulations COC c oncentration

- of

- c oncern DCI data call

- in EC 50 m edian (or 50 percent) e ffect c oncentration ECOTOX ECOTOXicology EDI e stimated daily i ntake EDSP E ndocrine Disruptor Screening Program E

- FAST Exposure and Fate Assessment Screening Tool EP end

- use product EPI Suite Estimation Program Interface Suite EPA Environmental Protection Agency FDA Food and Drug Administration FFDCA Feder al Food, Drug, and Cosmetic Act FIFRA Federal Insecticide, Fungicide, and Rodenticide Act FQPA Food Quality Protection Act FWP Final Work Plan g/mol g rams per mole GLN guideline number H 2 S hydrogen sulfide HHT hexahydro

- 1,3,5

- tris (2

- hydroxyethyl)

- s

- triazine Koc o rganic carbon normalized soil

- water partition coefficient Kow o ctanol

- water partition coefficient LC 50 m edian (or 50 percent) l ethal c oncentration LD 50 m edian (or 50 percent) l ethal d ose LOAEC l owest

- o bserved

- a dverse

- e ffect

- c oncentration LOAEL lowest

- observed

- adverse

- e ffect

- l evel Log K ow l ogarithm of the octanol

- water partition coefficient
μg m icrogram ml/g m illiliter p er g ram mg/kg m illigram p er k ilogram mg/kg/day m illigram p er k ilogram p er d ay mg/L m illigram p er l iter mm Hg m illimeter of mercury MOE m argin of e xposure MP manufacturing

- use product MRID Master Record Identification Number MRL m aximum r esidue l imit N/A n ot a pplicable

nm n anometers NOAEC no

- observed

- adverse

- effect

- c oncentration NOAEL n o

- o bserved

- a dverse

- e ffect

- l evel OCSPP Office of Chemical Safety and Pollution Prevention OECD Organization for Economic Co

- operation and DevelopmentDocket Number EPA

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P age 4 of 74 OPP Office of Pesticide Programs PC Code Pesticide Chemical Code pH power of h ydrogen or power of the con centration of the hydrogen ion pKa p ower of the acid dissociation constant or negative base

- 10 logarithm of the acid

dissociation constant of a solution ppb p arts p er b illion
ppm p arts p e r m illion PWP Preliminary Work Plan QSAR q uantitative s tructure

- a ctivity r elationship RED Rere gistration Eligibility Decision SAR s tructure a ctivity r elationship SF s afety f

actor TGAI t echnical grade active i ngredient TMDL total maximum daily l oads UF uncertainty factor UV/VIS ultraviolet/v isible light a bsorption WWTPs w astewater treatment plantsDocket Number EPA

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### Table of Contents
1 Introduction

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

................................ ................................ ................. 7 1.2 Updates t o the Workplan

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

................................ ................................ ................................ ................. 8 1.4 Chemical Identification and Properties

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

................................ ................................ ................................ ... 11 1.5.1 Registrations

................................ ................................ ................................ ........... 11 1.5.2 Summary of Registered Uses

................................ ................................ .................. 12 1.5.4 Usage Information

................................ ................................ ................................ .. 13 1.6 Regulatory History

................................ ................................ ................................ ......... 14 1.6.1 Tolerance Information

................................ ................................ ............................ 14 1.7 Incidents

................................ ................................ ................................ ......................... 14 1.7.1 Human Health

................................ ................................ ................................ ......... 14 1.7.2 Ecological

................................ ................................ ................................ ............... 14 2 Anticipated Data Needs

................................ ................................ ................................ ........ 14 3 Human Health Risk Assessment

................................ ................................ ........................... 20 3.1 Existing Toxicological Endpoints

................................ ................................ .................. 20 3.2 Dietary Exposure

................................ ................................ ................................ ............ 21 3.2.1 Food

................................ ................................ ................................ ........................ 22 3.2.2 Drinking Water

................................ ................................ ................................ ....... 23 3.3 Occupational and Residential Exposures

................................ ................................ ....... 23 3.3.1 Occupational Handler Exposure

................................ ................................ ............. 23 3.3.2 Occupational Post

- application Exposures

................................ ............................... 24 3.3.3 Residential Handler

................................ ................................ ................................ . 25 3.3.4 Residential Post

- Application Exposures

................................ ................................ . 26 3.4 Aggregate and Cumulative Exposure

................................ ................................ ............. 27 3.4.1 Aggregate Exposures

................................ ................................ .............................. 27 3.4.2 Cumulative Exposures

................................ ................................ ............................ 27 4 Environmental Risk Assess ment

................................ ................................ .......................... 27 4.1 Environmental Fate Assessment

................................ ................................ .................... 27 4.1.1 Water Quality

................................ ................................ ................................ .......... 28 4.2 Conceptual Models for Environmental Exposure Pathways

................................ .......... 28 4.3 Ecological Effects Assessment

................................ ................................ ....................... 29 4.3.1 Mechanism of Action

................................ ................................ .............................. 29 4.3.2 Measures of Eff

ect (Ecotoxicology Endpoints)

................................ ...................... 29 4.4 Exposure Analysis Plan

................................ ................................ ................................ .. 30 4.4.1 Aquatic and Terrestrial Wildlife Exposure Estimates

................................ ............ 30 4.4.2 Screening Level Down

- the

- Drain Analysis

................................ ............................ 31 4.5 Effects Analysis Plan

................................ ................................ ................................ ..... 31 5 Endocrine Disruptor Screening Program (EDSP)

................................ ................................ 31 6 Label Changes

................................ ................................ ................................ ....................... 32 7 Next Steps

................................ ................................ ................ 33 8 References

................................ ................................ ................ 34

Appendix A T oxicology Profile

................................ ................................ .............................. 35Docket Number EPA

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P age 6 of 74 Appendix B Environmental Fate

................................ ................................ ............................. 60 Appendix C Ecotoxicology Profile

................................ ................................ .......................... 63 Appe ndix D Screening Level Down

- the

- Drain Analysis

................................ ........................ 69 Appendix E Product Chemistry

................................ ................................ ............................... 70 Appendix F Agency Response to Public Comments

................................ .............................. 72

List of Tables Table 1

– Anticipated Risk Assessments and Data Needs for Registration Review: HH T

............. 9 Table 2

– Anticipated Registration Review Schedule

................................ ................................ .. 10 Table 3

– Chemical Identification of HHT

................................ ................................ ................... 10 Table 4

– Phy sical

- Chemical and Environmental Fate Properties for HHT

................................ . 11 Table 5

– Summary of HHT Registered Uses

................................ ................................ .............. 12 Table 6

-- Risk Mitigation Measures Specified in the HHT RED

................................ ................. 13 Table 7

– Studies Anticipated as Needed for the Registration Review of HHT

........................... 15 Table 8

– Data Required by the May 12, 2010 Data Call

- In for the HHT RED

.......................... 18 Table 9

– Existing Toxicological Endpoints for HHT

................................ ................................ .. 20 Table 10

– Occupational Handler Exposure Scenarios for HH T

................................ .................. 24 Table 11

– Occupational Post

- application Exposure Scenarios for HHT

................................ ..... 25 Table 12

– Residential Handler Exposure Scenarios for HHT

................................ ..................... 25 Table 13

– Residential Post

- application Exposure Scenarios for HHT

................................ ........ 26 Table 14

– Ecological Effects Endpoints Selected for Assessing Risks from HHT Use

............. 29Docket Number EPA

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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 hexahydro

- 1,3,5

- tris (2

- hydroxyethyl)

- s

- triazine

, also herein referred to as HHT

. The FWP document explains what EPA's Office of Pesticide Programs (OPP) knows about HHT , highlighting anticipated data and assessment needs, identifying the types of information that would be esp ecially useful to the Agency in conducting the review, and providing an anticipated timeline for completing HHT ’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 HHT and what types of new data or other information would be helpful for the Agency to receive as it moves toward a decision on HHT

.

T he d ocuments associated with HHT’s regis tration review can be viewed at http://www.regulations.gov in docket number EPA

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- 0654 . T he antimicrobial agent in HHT is formaldehyde (HCHO); therefore, this regi stration review also addresses potential risks associated with formaldehyde exposure from HHT uses. Products containing formaldehyde as an active ingredient will be evaluated under registration review case 0556. EPA’s formaldehyde Reregistration Eligibilit y Decision ( RED ) is available in the formaldehyde public docket at http://www.regulations.gov in docket number EPA

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## 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 continu e 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 re evaluates 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 A gency 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 s tandard for registration. The regulations governing registration review begin at 40 CF R 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 fo r 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.Docket Number EPA

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## 1.2 Updates to the Workplan
Since the publi cation of the PWP, the Agency has made the following updates:

 Revised Table 1 and Table 7 , Anticipated Data Needs, to include an assessment for existing residential uses and additional data needed to assess uses not anticipated at the time of the PWP

. Added the following risk asse ssment data needs: metabolism and pharmacokinetics, dermal exposure for occupational handler and post

- application, i nhalation e xposure for occupational handler and residential post

- application

, p roduct

u se, nature of residue on surfaces, and migration stud y

.

 Added a c tivated sludge sorption isotherm as a data requirement to Table 7.

 Added acute neurotoxicity study as a data requirement triggered both by indirect food residue > 200 ppb and metal working fluid use.

 Updated Table 2, Anticipated Registration Rev iew Schedule.

 Revised Table 4, vapor pressure information.

 Remove d Footnote 1 Table 8

.

 Updated Section 1.5.2, Su mmary of Registered Uses, to include existing residenti al uses.

 Added use information from the Kline report to Section 1.5.4

.

 Revised Section 3.2 , Dietary, to assess risks associated with EPA Reg. No . 68868

- 1

.

 Revised Section 3.3 and 3.4 , to incorporate u ses previously not included in PWP

.

 Revised Section 6, label changes.

 Removed Section 7, Guidance for Commenters.

 Updated Section 8, Next Steps (now Section 7).

 Updated Table E1

 Added Appendix F, Agency Response to Public Comments.

 Included a list of references for all Appendices.

The Agency received 2 submissions d uring the public comment period on the initial docket. See Appendix F for a summary of the comments and the Agency’s response

. 

## 1.3 Case Overview
The Preliminary Work Plan (PWP) for HHT was published on December 1 9 , 2014 and the 60

- day comment period ended on F ebruary 17, 2015. Public comments received concerning the PWP and d ocuments associated with this regis tration review can be vie wed at http://www.regulations.gov in docket number EPA

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.
Tables 1 and 2 b elow summar ize the issues relevant to this registration review case and the anticipated registration review schedule

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P age 9 of 74 Table 1

–Anticipated Risk Assessments and Data Needs for Registration Review : HHT

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 7 for details)

Dietary (food) Yes 6/30/2008 New Nature of Residue Data Migration Study Immunotoxicology Acute Neurotoxicity Dietary (drinking water) Yes N/A New None (See section 3.2.1 )

Occupational Handler Yes 6/30/2008 Updated Immunotoxicity Acute Neurotoxicity Indoor Inhalation Exposure Indoor Dermal Exposure Product Use

Occupational Post Application Yes 6/30/2008 Upda ted Indoor Inhalation Exposure Indoor Dermal Exposure Product Use

Residential Handler Yes 6/30/2008 Updated Indoor Inhalation Exposure Indoor Dermal Exposure Product Use Acute Neurotoxicity

Resident i al Post Application

- Incidental Oral and Dermal Yes 6/30/2008 Updated Indoor Dermal Exposure Indoor Surface Residue Dissipation Product Use Acute Neurotoxicity

Resident i al Post Application

- Inhalation Yes 6/30/2008 Updated Inhalation Exposure Product Use

Aggregate Yes N/A New Nature of Residue on Surfaces Migration Study Immunotoxicology Acute Neurotoxicity

Cumulative No (See section 3.3.2 ) N/A None None

Tolerance Review No (See section 1.6.1 ) N/A None None

Ecological Yes 6 / 30 /2008 Updated Vascular plant growth Algae growth Fish early life stage Aquatic invertebrate life cycle Hydrolysis Aerobic aquatic metabolism Adsorption/desorption Waste water treatment plant studies 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 8

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P age 10 of 74 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 Preli minary Work Plan 2014

- 12 2014

- 12

- 1 9 Close Public Comment Period 2015

- 02 2015

- 02

- 17 Phase 2: Case Development Issue Final Work Plan 2015

- 06 2015

- 06 Issue Data Call

- In (DCI) 2016

- 06 Receive Data to be Considered in Risk Assessment 2018

- 06 Open 30

- Day Public Comment Period for Preliminary Risk Assessment(s) 2019

- 12 Close Public Comment Period 2020

- 01 Phase 3: Registration Review Decision and Implementation Open 60

- Day Public Comment Period for Pro posed Decision 2020

- 06 Close Public Comment Period 2020

- 08 Issue Final Decision 2020

- 12 Begin Post

- Decision Follow

- up 2020 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 3074: HHT (PC Code 083301 )

. Note: HHT i s also a formaldehyde releaser. The remaining compound after the release of formaldehyde is 1,3,5

- trimethyl triazine.

Table 3

– Chemical Identification of HHT

Chemical Name HHT
Chemical Classification Cyclic triazines
PC Code 083301
CAS N umber 4719

- 04

- 4
Molecular Formula C 9 H 21 N 3 O 3 Molecular Weight (grams/mole) 219.29 g/mol

Molecular Structure HHT Formaldehyde
CH2O 1,3,5 trimethyl triazine:

The HHT product chemistry information relevant to risk assessment is summarized in Table 4

, and the details of the product chemistry and environmental fate information are discussed in Appendix E and B, respectively

. The Agency has no existing environmental fate data on theDocket Number EPA

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P age 11 of 74 degrada te 1,3,5 trimethyl triazine ; studies are anticipated to be required for the registration review risk assessment (see table 7 for a list of anticipated studies on this degradate ).

Table 4

– Physical

- Chemical and Environmental Fate Properties for HHT

Guideline No. P arameter HHT

Physical/Chemical Properties
830.7000 pH NA
830.7050 UV/Visible Absorption NA
830.7200 Melting point 155 0 C

830.7550 Octanol

- water partition coefficient (Log K ow ) - 2.3 (pH 5, 24 0 C),

- 2.0 (pH 7, 24 0 C);

- 1.3 (pH 9

, 24 0 C); ( MRID # 47284308)
830.7840 Solubility in water 3 g/L (EPI Suite, version 4.11)

830.7950 Vapor pressure 1 4.1 x 10

- 9 mm Hg at 25 C (EPI Suite, Web version 1.0 ) Mobility Organic Carbon Partitioning Coefficient Log K oc 10 L/kg (EPI Suite, version 4.11) Persistence in Water (half

- life)

Hydrolysis 2 at 25 o C: Half

- life at pH 5: 3.2 days; pH 7: 50 days; pH 9: 302 days (MRID# 43181001)
Hydrolysis at 37 o C pH 4, half

- life = 0 min; pH 11.35 , no degradation ; (MRID# 47353803) 1. The previously reported vapor pressures ( 11.2 mm Hg, at 20 0 C ; 15.15 mm Hg at 25 0 C; 61.1 mm Hg at 50 0 C) ; are from a study (MRID # 47284307) measuring for a solution containing 79% HHT and 18% H 2 0. These measurements represent the vapor pressure of H 2 0 rather than HHT. 2

. A hydrolysis study conducted with metalworking fluids was also conducted and is discussed further in Section 4.1. The hydrolysis in this study resulted in the loss of a significant portion of the material which lim ited the utility of the study.



## 1.5 Use/Usage Description

1.5.1 Registrations

There are 11 EPA

- registered products that contain HHT as an active ingredient (a.i.) and used as

a material preservative, bactericide and fungi cide in coolants , and for oilfield uses

. The p ercent a.i. for end

- use products ranges from 39.3 percent to 78.5 percent , and the formulations include soluble concentrate, ready

- to

- use solution, flowable concentrate, emulsifiable concentrate, and wettable powder. Three of these products contain o ther active ingredients such as sodium pyrithione or 1, 2

- benzisothiazolin

- 3

- one . There is one manufacturing use product (EPA Registration Number 1839

- 59 ) that contains 7 8.5 percent hexahydro

- 1,3,5

- tris (2

- hydroxyethyl)

- s

- triazine

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### 1.5.2 Summary of Registered Us es
Table 5 presents a summary of the registered uses of HHT that will be assessed for registration review. HHT end use products can be applied by liquid pour, liquid pump and solid pour wettable powder

. Household cleaning products preserved with HHT can be applied by aerosol can, trigger sprayer, mop and wipe and paints preserved with HHT can be applied by brush/roller and airless spray. Several existing uses, application methods and application rates were considered ineligible for reregistration at the time of the RED. Subsequently, registrants have indicated their intent to provide information to help the Agency refine risk identified during the RED process. Table 5 has been updated to represent all registered uses

.

Table 5

– Summary of HHT Registered Uses

Use Application Method Application Rate 2 Material Preservative Coatings, inks, dyes and slurries Liquid Pour , Liquid Pump 1100 ppm

Adhesives, dispersions, and emulsions Solid Pour Wettable Powder (WP) 2358 ppm

Liquid Pour, Liquid Pump 1650 ppm

Dish Detergent 1 Liquid Pour 1550 ppm

Hard Surface Cleaners 1 Liquid Pour A erosol C an, T rigger S prayer M op and W ipe 1550 ppm

Hydrogen Sulfide Scavenger Systems Liquid Pour, Liquid Pump 6.44 lb ai/lb H 2 S

Laundry Detergents 1 Liquid Pour 1550 ppm

Fuel Oils Liquid Pour, Liquid Pump 1180 ppm

Paint 1 Liquid Pour, Liquid Pump Brush/roller and Airless Spray 2355 ppm

Reagents Used in Medical Fluids Liquid Pour, Liquid Pump 393 ppm
Metal working fluids Liquid Pour, Liquid Pump 500 ppm
Metalworking fluids 1 Liquid Pour, Liquid Pump 1100 ppm Industrial Processes and Water Systems

Coolants Liquid Pump, Liquid Pump 1180 ppm

Oilfield Uses Solid Pour WP 1200 ppm
Liquid Pump, Liquid Pour 936 ppm

1 These uses were not included in the preliminary work plan because the A gency expected all registrants to comply with the mitigation specified in the RED, including the removal of uses and the reduction of application rates. The registrant of the last remaining product with uses that were ineligible for reregistration has indi cated its intent not to add the mitigation measures specified in the RED but to now support the uses; therefore, the currently

- registered uses that were found to be ineligible for reregistration as well as the related newly anticipated data needs and relat edDocket Number EPA

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P age 13 of 74 newly anticipated risk assessments have been added to this work plan. The Agency will also continue to pursue label compliance through regulatory or other action during this registration review.

2 Only maximum application rates are captured.

1.5.3 Summary of Risk Mitigation Measures from HHT RED

Table 6 presents the mitigation measures outlined in the 2008 RED and corrects a typographical error from parts per billion (ppb) to parts per million (ppm).

Table 6

-- Risk Mitigation Measures Specified in the HHT RED Mitigation Measures for HHT Use Site Risk(s) of Concern Mitigation Measures

Paints, Stains, and Coatings Residential handler inhalation exposure to HHT in paint Delete/cancel use Occupational handler inhalation exposure to formaldehyde in paint

Metal Working Fluids Occupational handler inhalation exposure to formaldehyde in metal working fluids Reduce maximum application rate from 1600 ppm to 500
ppm 1

Institutional and Household Cleaning Products Residential handler inhalation exposure to formaldehyde in cleaners. Residential handler inhalation exposure to HHT in laundry detergent Delete/cancel use

Oil Field Secondary Recovery Occupational handler dermal exposure to HHT in oil field secondary recovery uses Reduce maximum application rate from 0.16% active ingredient (ai) to 0.12% a.i.

1 Error in the RED: Units should be parts per million (ppm), not ppb.



### 1.5.4 Usage Information According to the Specialty Biocides 2004

- 2005 Report (Kline, 2006)

, the estimated consumption of triazines, which include HHT, was 8,100,000 pounds in 2004. The m etal

w orking f luid ( MWF ) use for the triazines accounted for 72 percent of this consumption, followed by paints (18.5%), adhesives (7.4%) and latex polymers (3.0%). The most current use information available will be used for the registration review risk assessment.Docket Number EPA

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## 1.6 Regulatory Histor y

The first product containing HHT as an active ingredient was r egist ered in the United States in 197 2. The Agency completed a RED for HHT in 200 8 . The post

- RED DCI was issued on May 12, 2010

. See Section 2 for a review of the status of the data required by the post

- RED DCI

.


### 1.6.1 Tolerance Information EPA has not established tolerances in raw agricultural commodities or processed food and feed products for HHT under the Federal Food, Drug and Cosmetic Act (FFDCA) Section 408. There are also no tolerance exemptions for HHT under FFDCA Section 408. HHT ha s not been cleared as a food additive by the US Food and Drug Administration (US FDA) under FFDCA Section 409

. 

## 1.7 Incidents

1.7.1 Human Health

In the case of HH T, there are three aggregate incidents listed in the IDS for the year s 2008 to October 3 0 , 2014

. Two of these incidents are classified as cannot be rated (HE) ; no details were included . The remaining incident was classified as human minor (HD) . Sore throat, watery eyes, respiratory irritation

, and burning of the lungs w ere experienced two weeks after occupational
exposure to the aqueous product

.


### 1.7.2 Ecological No HHT incidents have been reported in the Agency ’s Ecological Incident Information System (EI I S) as of April 15 , 201 5

. 

## 2 A nticipated Data Needs

The Agency anticipates reviewing any data recei ved in response to the post

- RED DCI as well as data requested for this registration review prior to conducting the registration review risk assessments for HHT

. Table 7 presents a summary of the data anticipated as being needed to support this registration review

, and Table 8 presents a summary of the status of the post

- RED DCI issued on May 12, 2010

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Page 15 of 74 Table 7

– Studies Anticipated as Needed for the Registration Review of HHT

GLN Study Name Test Substance 1 Time Frame (Measured in months from DCI Receipt) Risk Assessment(s)

Data Will Support Use Site(s) Triggering Anticipated Data Requirement Applicable

Exposure Scenario

835.2120 Hydrolysis Parent and degradate: 1,3,5

- trimethyl triazine 12 All Industrial Water Processes, Material Preservative Use All

835.4300 Aerobic aquatic

metabolism Parent and the degradate: 1,3,5

- trimethyl

- triazine 12 All Industrial Water Processes, Material Preservative Use All

835.1230 Adsorption/ desorption Parent and the d egradate: 1,3,5

- trimethyl

- triazine 12 All Industrial Water Processes, Material Preservative Use All

835.1100 2 Activated Sludge

Sorption Isotherm Parent and the degradate: 1,3,5

- trimethyl

- triazine 12 All Industrial Water Processes, Material Preservative Use Down the drain

835.3110 3
835.3220 3
835.3240 3
835.3280 3 Ready biodegradability

or one of three biodegradation in activated sludge simulation tests Parent and the degradate: 1,3,5

- trimethyl triazine 12 All Industrial Water Processes, Material Preservative Use Down the drain

850.3300 4 Activated sludge,

respiration inhibition test Parent and the degradate: 1,3,5

- trimethyl triazine 12 All Industrial Water Processes, Material Preservative Use Down the drain

__________________________ 1 Information on the degradate 1,3,5

- trimethyl triazine which remains after the release of formaldehyde may potentially be available from information on other triazines. The registrants could propose triazine data and bridging information if appropriate in lieu of guideline studies o n the degradate.

2 EPA has a published final guideline for this study: http://www.regulations.gov/#!documentDetail;D=EPA

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- 0003

. 3 The results of the Activated Sludg e, Respiration Inhibition Test (ASRI), GLN 850.3300, will determine which of the four biodegradation tests would be required.

 If the ASRI test results in an EC50 of less than or equal to 20 mg/L, then either the (i) Simulation tests to assess the biodegra dability of chemicals discharged in wastewater, GLN 835.3280, (ii) the Simulation Test

– Aerobic Sewage Treatment

- – A. Activated Sludge Units, GLN 835.3240, or (iii) the Porous Pot Test, GLN 835.3220 would be required.

 If the ASRI test results in an EC50 greater than 20 mg/L, then the registrant would be required to conduct either the (i) Ready Biodegradability, GLN 835.3110, (ii) Simulation tests to assess the biodegradability of chemicals discharged in wastewater, (iii) the Simulation Te st

– Aerobic Sew age Treatment

– A. Activated Sludge Units, or (iv) the Porous Pot Test.

 If the Ready Biodegradability Study is conducted and passes, then no further testing would be required. If, however, the pesticide fails the Ready Biodegradability study, then the (i) Simulation tests to assess the biodegradability of chemicals discharged in wastewater, (ii) the Simulation Test

– Aerobic Sewage Treatment

– A. Activated Sludge Units, or (iii) the Porous Pot Test would be required

. 4 EPA published draft guidance under gu ideline 850.6800 and has since published final guidance for this study under guideline 850.3300: http://www.regulations.gov/#!documentDetail;D=EPA

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- 0021 . OECD Test Guideline 209 can also be used as guidance for this study, available online at http://www.oecd

- il ibrary.org/content/book/9789264070080

- en

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P age 16 of 74 GLN Study Name Test Substance 1 Time Frame (Measured in months from DCI Receipt) Risk Assessment(s)

Data Will Support Use Site(s) Triggering Anticipated Data Requirement Applicable Exposure Scenario

850.1300 Aquatic invertebrate

life cycle TGAI 2 4 Ecological Assessment Industrial Water Processes, Material Preservatives Down the drain

850.1400 Fish early life stage TGAI 2 4 Ecological Assessment Industrial Water Processes, Material Preservatives Down the drain

850.4400 Aquatic vascular plant

growth TGAI 12 Ecological Assessment Industrial Water Processes, Material Preservatives Down the drain

850. 45 00 4 Aquatic algal growth TGAI 12 Ecological Assessment Industrial Water Processes, Material Preservatives Down the drain

850.4550 4 Algal Toxicity Tier II TGAI 12 Ecological Assessment Industrial Water Processes, Material Preservatives Down the drain

870. 6 200 Acute

N eurotoxicity

—

rat. TGAI 12 Human Health Assessment Countertop Disinfection ;

Dish Detergents; Metal

- working Fluids Commercial settings ; Oc cupational machinists

870.7485 Metabolism and

Pharmacokinetics TGAI 12 Human Health Assessment All All

870.7800 Immunotoxicity TGAI 12 Human Health Assessment All All

875.1200 Dermal Exposure EP 24 Occupational Handler Material Preservatives Machinist Brush/Roller Airless Sprayer Mop Spray and Wipe Liquid Pour Solid Pour WP

875. 1400 Inhalation Exposure EP Degrad ate: formaldehyde 24 Occupational Handler Material Preservatives Machinist Brush/Roller Airless Sprayer Mop Spray and Wipe Liquid Pour Solid Pour WP

875.1700 Product Use EP 1 2 Occupational Handler Material Preservatives AllDocket Number EPA

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P age 17 of 74 875.2400 Dermal Exposure EP 2 4 Occupational Post

application Manufacturing Settings of

Material Preservatives Post

- application

875.2500 5 Inhalation Exposure EP Degradate: formaldehyde 24 Occupational and Residential Post

application Manufacturing Settings of

Material Preservatives Post

- application Exposure Bystanders

None Nature of Residue on

Surfaces 6 TGAI 12 Dietary (food) Countertop Disinfection Dish Detergents Residential and Commercial Post

application

None Migration Study 7 EP 12 Dietary (food) Countertop Disinfection Dish Detergents Residential and Commercial Post

application

__________________________ 4 On June 27, 2012, EPA announced certain revisions in harmonized guideline series 850

– Ecological Effects Tests

– including renumbering and other designations or changes for some guideline studies. See “Final Test Guidelines; O CSPP 850 Series; Notice of Availability” 77 FR 38282, June 27, 2012. http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

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. 5 Formaldehyde inhal ation exposures are expected to occur to worker bystanders as a result of HHT material preservative applications in industria l settings. Currently, no data are available to assess these worker bystander exposures

, and therefore, monitoring data are anticip ated to be needed. 6 If an antimicrobial pesticide may be applied to a food

- contact surface or impregnated into a food

- contact material and if theoretical (high

- end) estimates of exposure exceed EPA's risk level of concern, then the nature of the residue o n surfaces study would be expected to be required. The anticipated DCI will provide that p rotocols must be approved by the Agency prior to the initiation of the study.

7 Based on the results of the nature of the residue on surfaces study, if residues of concern are identified, then the migratio n study would be expected to be required and p rotocols must be approved by the Agency prior to the initiation of the study.Docket Number EPA

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Page 18 of 74 Table 8

– Data Required by the May 12, 2010 Data Call

- In for the HHT RED

GLN Study Name Test Substance Risk Assessment(s)

Data Will Support Use Site(s) Triggering Anticipated Data Requirement Applicable Exposure Scenario Status

870.1300 Acute inhalation toxicity TGAI, MP Occupational and Residential Handlers and P ost

- applications All Human Health Exposure Unsatisfied

870.3150 90

- day oral toxicity in non

rodents TGAI Occupational and Residential Handlers and P ost

- applications ; Drinking Water All Human Health Exposure Unsatisfied

870.3465 90

- day inhalation toxicity TGAI Occupational and Residential Handlers and P ost

- applications All Human Health Exposure Unsatisfied

870.3700 Prenatal developmental toxicity

study in non

- rodent TGAI Occupational and Residential Handlers and P ost

- applications ; Dietary (Food and Drinking Water ) All Human Health Exposure Unsatisfied

870.3800 Reproduction and fertility

effects TGAI Occupational and Residential Handlers and P ost

- applications ; Dietary (Food and Drinking Water ) All Human Health Exposure Unsatisfied

870.4100 Chronic toxicity TGAI Occupational and Residential Handler and P ost

- applications ; Dietary (Food and Drinking Water ) All Human Health Exposure Unsatisfied

870.4200 Carcinogenicity TGAI Occupational and Residential Handler and P ost

- applications ; Dietary (Food and Drinking Water ) All Human Health Exposure Unsatisfied

875.1100 Dermal exposure

-- outdoor EP N/A N/A N/A Waived 1

875.1200 Dermal exposure

-- Indoor EP Occupational Handler Material Preservatives; Industrial Processes and Water Systems Pour Liquid

Pour Solid Unsatisfied

875.1300 Inhalation exposure

-- outdoor EP N/A N/A N/A Waived 1

875.1400 Inhalation exposure

-- indoor EP Occupational Handler Material Preservatives; Industrial Processes and Water Systems Pour Liquid

Pour Solid Unsatisfied

875.1600 Application exposure

monitoring data reporting EP N/A N/A N/A Waived 2Docket Number EPA

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P age 19 of 74 GLN Study Name Test Substance Risk Assessment(s)

Data Will Support Use Site(s) Triggering Anticipated Data Requirement Applicable Exposure Scenario Status

875.1700 Product Use Information EP Occupational Handler Material Preservatives; Industrial Processes and Water Systems Pour Liquid

Pour Solid Unsatisfied

875.2700 Product Use Information EP Occupational Post

- application; Occupational Bystander Metalworking Fluids; Oil Field Occupational Post

- application;

Occupational Bystander Unsatisfied

875.2800 Description of human activity EP Occupational Post

- application; Occupational Bystander Metalworking Fluids; Oil Field Occupational Post

- application;

Occupational Bystander Unsatisfied

875.2900 Data reporting and calculations EP N/A N/A N/A Waived 2 *N/A = not applicable

1

. Only indoor exposure data are required. 2

. The Agency will waive the data reporting studies (GLN 875.1600 and 875.2900) as the data requirement is now incorporated in the dermal or inhalation exposure study.Docket Number EPA

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Page 20 of 74 

## 3 Human Health Risk Assessment

The Agency anticipates the need to conduct a human health risk assessment for HHT and its d egradate

, formaldehyde

, upon receipt of the nu merous data studies required under the post

- R ED DCI

. Additionally, the Agency anticipates the need to require additional data f or currently registered use s under registration review

. These additional studies include acute neurotoxicity, immunotoxicity

, metabolism/pharmacokinetics, and dermal and inhalation exposure stud ies (as noted in Table 7 ).

The Agency is currently reevaluating the carcinogenic potential of form aldehyde and completing

a new Integrated Risk Information System ( IRIS ) assessment and unit risk value for formaldehyde. The Agency plans to use this revised assessment, once the peer review process is completed, to evaluate the risk related to formaldehyde released as a result of the uses of HHT as

an antimicrobial pesticide.



## 3.1 Existing Toxicological Endpoints

The Agency anticipates the need to revise the existing toxicological endpoints , shown in Table 9
, as part of this registration review. All available information

, including existing toxicology studies and valid scientific literature, will be considered in establishing the toxicology endpoints to be used in the registration review risk assessment. Additional studies, as outlined in Table 7
, are anticipated to be required, particularly for registered uses that may result in dietary exposure

. A detailed description of the available toxicity studies is provided in Appendix A.

Table 9

– Existing Toxicological Endpoints for HHT

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

Dietary Risk Assessment Acute Dietary (All populations) NOAEL maternal = 500 MOE = 1,000 (10x interspecies, 10x intraspecies, 10x database uncertainty) Developmental Toxicity

– Rat (MRID 41161801)

LOAEL maternal = 7 50 mg/kg/day , based on decreased body weight gain, ulcerations

, and scarring of the stomach mucosa
Acute RfD Acute RfD = 500 mg/kg/day ÷ 1000 = 0.5 mg/kg/day Chronic Dietary (All populations) NOAEL = 50 UF = 100 (10x

– Intra; 10x

– Inter) 90

- day Oral Study

– Rat ( MRID 41483001 )

LOAEL = 10 0 mg/kg/day , based on lymphocytic infiltration in females and erosion of gastric mucosa and prominence of the limiting ridge of the stomach in males
Chronic RfD Chronic RfD = 50 mg/kg/day ÷ 100 = 0.5 mg/kg/day Occupational/Residential (Non

- Dietary) Risk AssessmentDocket Number EPA

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P age 21 of 74 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

Incidental Oral Short

- Term (1

- 30 days) NOAEL maternal = 500 MOE = 1,000 (10x interspecies, 10x intraspecies, 10x database uncertainty) Developmental Toxicity

– Rat (MRID 41161801)

LOAEL maternal = 7 50 mg/kg/day , based on decreased body weight gain, ulcerations

, and scarring of the stomach mucosa Incidental Oral Intermediate

- Term (30 days

- 6 months) NOAEL = 50 UF = 100 (10x

– Intra; 10x

– Inter) 90

- day Oral Study

– Rat ( MRID 41483001 )

LOAEL = 10 0 mg/kg/day , based on lymphocytic infiltration in females and erosion of gastric mucosa and prominence of the limiting ridge of the stomach in males Dermal A bsorption Facto r Not required as a dermal toxicity study has been used for the dermal endpoint.

Dermal Short

- Term (1

- 30 days) No risk assessment necessary because n o adverse systemic effects observed in a 21

- day dermal

toxicity study up to 1

, 000 mg/kg /day.

Dermal I nter mediate

- term (30 days

- 6 months) and long

- term (>6 months) NOAEL systemic = 250 MOE = 1,000 (10x interspecies, 10x intraspecies, 10x database uncertainty) 90

- day dermal Study

– Rat (MRID 41483002 )
Systemic NOAEL was found to be greater tha n 250 mg/kg/day (the HDT); systemic LOAEL was not established. Inhalation (all durations) NOAEL = 50 MOE = 1000 (10x interspecies, 10x intraspecies, 10x database uncertainty) 90

- day Oral Study

– Rat (MRID 41483001)
LOAEL = 100 mg/kg/day, based on lymphocytic infiltration in females and erosion of gastric mucosa and prominence of the limiting ridge of the stomach in males



## 3.2 Dietary Exposure
EPA anticipates the need to conduct a dietary exposure (food and drinking water ) assessment to support registration review of HHT since no previous dietary assessments have been conducted and there are uses on one label that could result in indirect food contact and dietary exposure to HHT (EPA Reg. No. 68868

- 1). Also, t he Agency ant icipates the need to determine the potential for discharged residues of HHT to result in human dietary exposure via drinking water, which may trigger a dietary ( drinking water ) assessment during Registration Review

. Dietary risk is characterized in terms of the Population Adjusted Dose (PAD) which reflects the reference dose (RfD), either acute or chronic, that has been adjusted to account for the FQPA Safety Factor (SF). This calculation is performed for each population subgroup for which an endpoint exis ts. A risk estimate that is less than 100% of the acute or chronic PAD is not of concern. Acute and chronic dietary risk for HHT will be assessed by comparing dietary exposure estimates expressed in mg/kg/day to the acute Population Adjusted Dose (aPAD) an d chronic Population Adjusted Dose (cPAD).Docket Number EPA

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P age 22 of 74 

### 3.2.1 Food

D ietary exposure assessments will be conducted during registration review since c urrent ly registered uses of HHT may result in dietary (food) exposure . A screening

- level (food only) analysis was conducted for this work plan using established toxicological points of departure (PODs), inputs from EPA Reg. No. 68868

- 1, and an active ingredient concentration of 1570 ppm for hard surface cleaners in residential and commercial settings and dish detergents using the screening

- level Food and Drug Administration (FDA) model for food contact (hard surfaces) in commercial and residential areas

. The acute population adjusted dose (aPAD) is 0.5 mg/kg/day and the chronic population adjusted dose (cPAD) is 0.5 mg/kg/day. Howe ver, since the aPAD and the cPAD are the same, and the inputs/consumption data for the dietary exposure assessment are the same for both durations, the dietary exposure assessment outlined below is protective of both durations.

Dietary Exposure Assessmen t

- Residential Areas

The results of the screening

- level analysis for dish detergents and hard surface cleaners in residential areas do not result in risks of concern for the US population or any population subgroup over both acute and chronic durations (< 100% of the PAD) when using the screening

level FDA model.

For residential scenarios assuming a countertop surface area of 2000 cm 2 , the screening

- level Food and Drug Administration (FDA) model for food contact (hard) surfaces results in an total daily di etary intake of HHT exceeding 200 ppb for all infants <1 year old and children 1

- 2 years old. However, when calculating the total daily dietary intake using the Indirect Dietary Residential Exposure Assessment Model (IDREAM), which is a refined Tier II mod el for uses in hard surface cleaners, the refined EDI becomes < 200 ppb.

Dietary Exposure Assessment

- Commercial Areas

The results of the screening

- level analysis for the registered hard surface cleaners used in commercial settings indicate that the indi rect food uses of HHT result in risks of concern for all infants < 1 year old (>100% of the PAD). For commercial scenarios assuming a countertop surface area of 4000 cm 2 , the screening

- level FDA model for food contact (hard) surfaces results in an total da ily dietary intake exceeding 200 ppb for all infants <1 year old, children 1

- 2 years old, and children 3

- 5 years old.

Conclusions & Data Requirements

The screening

- level analysis for hard surface cleaners using the screening

- level FDA model for food conta ct (hard surfaces) in commercial areas results in risks above the Agency’s level of concern for some population subgroups. Additionally, the total daily dietary intake in residential and commercial areas is > 200 ppb for some population subgroups.

There fore, under 40 CFR 158.2230, total daily dietary intakes greater than 200 ppb trigger additional (Tier II) toxicity data that should be submitted to the Agency. Additional residue data (nature of residue on surfaces and migration study) are anticipated to be required because theDocket Number EPA

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P age 23 of 74 level of concern is exceeded (for two population subgroups) using the screening

- level assessments, per 40 CFR 158.2290.

If the toxicological PODs change during registration review, additional residue chemistry or toxicity data may b e required. Additionally, if current labels are amended during registration review to limit the use pattern to non

- food contact areas only, a dietary (food) exposure assessment would not be needed.

The dietary exposure contribution from the currently regi stered uses of HHT will be fully considered in the aggregate assessments conducted during registration review.



### 3.2.2 Drinking Water A drinking water assessment has not been conducted for HHT and its primary triazine degradate

, in the past. Howe ver, the Agency cannot rule out the potential for HHT to enter drinking water

. There are uncertainties with respect to the residues of concern in drinking water, the magnitude of these potential residues concern, their mammalian toxicities, how persistent they will be, a nd if they are likely to pass through a wastewater treatment plant (WW TP) is unknown. Therefore, the Agency anticipates needing certain environmental fate data listed in Section 4 of this F WP

. These data will be useful to determine the extent residues of H HT and its primary triazine degradate pass through a W WTP. The Agency anticipates using these data and/or other sources of information during the risk assessment stage of registration review to determine the potential for discharged residues of HHT to resu lt in human dietary exposure via drinking water, which may trigger a drinking water asse ssment

. 

## 3.3 Occupational and Residential Exposures

The Agency anticipates the need to update the occupational and residential exposure (ORE)
assessment done for the 2008 RE D during registration review. The assessment from the RED will need to be re vised to account for scenarios that were identified after the preliminary workplan (PWP) comment period and to utilize the exposure data anticipated to be received from the post

- RED and registration review data call

- ins

.


### 3.3.1 Occupational Handler Exposure EPA anticipates the need to revise the occupational handler assessment conducted in support of the 2008 RED (D346240). Occupational handler dermal and inhalation exposures to HHT were assessed in the RED for the following applications : aerosol can, airless sprayer, brush/roller, liquid pour, liquid pump, mop, spray and wipe, and solid pour. The Agency expects to conduct new assessments for these scenarios in registration review upon re ceipt of the anticipated exposure data expected from the post

- RED DCI.

Several existing uses, application methods (e.g., aerosol can, airless sprayer, brush/roller, mop, and spray and wipe) and application rates were considered ineligible for reregistration at the time of the RED. The use patterns included: 1) paints, stains and coatings; 2) institutional andDocket Number EPA

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P age 24 of 74 household cleaning products; and 3) metal working fluids that were above a concentration of 500
ppm. The registrant of the last remai ning product with uses that were ineligible for reregistration has indicated its intent not to add the mitigation measures specified in the RED but to now support the uses; therefore, the currently

- registered uses that were found to be ineligible for
rereg istration as well as the related newly anticipated data needs and related newly anticipated risk assessments have been added to this work plan. The Agency will also continue to pursue label compliance through regulatory or other action during this registra tion review

.

Table 10 lists the occupational handler scenarios that will be needed in registration review.

Table 10

– Occupational Handler Exposure Scenarios for HHT

Scenario Exposure Rou te(s) Duration
Occupational Exposures Liquid Pour for Material Preservatives and Industrial Processes and Water Systems Dermal Inhalation Short and Intermediate

Term Solid Pour Wettable Powder for Material Preservatives and Industrial Processes and Water Systems Dermal Inhalation Short and Intermediate

Term
Application of Paint by a Professional Painter (Airless Sprayer) Dermal Inhalation Short and Intermediate

Term
Application of Paint by a Professional Painter (Brush/Roller) Dermal Inhalation Short and Intermediate

Term Application of Commercial Household Cleaners (Spray and Wipe and Mop) Dermal Inhalation Short and Intermediate

Term Occupational Handler Inhalation Exposure to Formaldehyde in Metal Working Fluids and Oil Fields Inhalation Sho rt and Intermediate Term
Occupational Handler Exposure to Metal working Fluids (Liquid Pour) Dermal Inhalation Short and Intermediate

Term



### 3.3.2 Occupational Post

- application Exposures Metalworking Exposure

The RED assessed the dermal and inhalation exposure for machinists han dling treated metalworking fluids with HHT and the HHT degradate formaldehyde.

In the RED, the Agency assessed inhalation risks from HHT using a submitted inhalation monitoring study ( MRID 473354

- 02) which measured formaldehyde air conc entrations in several metalworking facilit ies

. The majority of the concentrations exceeded the EPA level of

concern (LOC) of 100 ppb . As a result, the Agency specified in the RED that the maximum application rates for HHT must be reduced from 1600 ppm to 500 ppm

. This rate reduction was based on a literature study (Linnainmaa, 2003) which indicated that 500 ppm was the minimum level needed to prevent microbial growth in MWF

. A r egistrant ha s indicated its intent to support concentrations above 500 ppm

.

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, the Agency received comments about the existence of a new metalworking exposure study that is expected to refine the Agency’s risk assessmentsDocket Number EPA

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P age 25 of 74 conducted during the RED. Based on the comments received during registra tion review, t he Agency expects to revise the existing dermal and inhalation assessments for machinists exposed to HHT and the degradate formaldehyde using the anticipated new metalworking exposure study cited in the public comment with data provided from the Antimicrobial Exposure Assessment Task Force (AEATF) II. The Agency expects the need to issue a data call

- in for th ese data during registration review.

Industrial Bystander Inhalation Exposure

Formaldehyde inhalation exposures are expected to occur to bystanders as a result of HHT material preservative applications in paints, commercial detergents and oil fields . Currently, no data are available to assess the bystander exposures and therefore, m onitoring data are expected to be needed. The data w ere not called in the post

- RED DCI , but the Agency anticipates the need for an inhalation chamber test s to fulfill the post

- application exposure data (Series 875.2500) in registration review for each of t hese uses

. Table 11 lists the scenarios that the Agency anticipates that will be need to be assessed in registration review for occupational post

- application
exposures

.

Table 11

– Occupational Post

- application Exposure Scenarios for HHT

Scenario Exposure Route(s) Duration
Occupational Exposures
Machinist Exposure to HHT and Formaldehyde Dermal Inhalation Chronic
Bystander Exposure to Formaldehyde ( P aints, detergents, and oil fields) Inhalation Chronic



### 3.3.3 Residential Handler

All of the HHT residential handler uses such as treated paints, cleaners and detergents were already assessed in the RED

, and the Agency determined these uses were ineligible for reregistration as a result of handler risk concerns. Subsequently, registrants have indicated their intent to provide information to allow the Agency to refine risk identified during registration review. T he Agency expects to revise the exis ting dermal and inhalation assessments for application of paints (airless sprayer, brush roller) and general cleaners (spray and wipe, and mop) to HHT and the degradate formaldehyde using the anticipated exposure study data provided from the Antimicrobial Exposure Assessment Task Force (AEATF) II. The Agency will issue data call

- ins for this exposure data during registration review and will conduct new residential handler risk assessments. Table 12 lists the residential handler exposure scenarios that the Agency anticipates will be need ed for registration review.

Table 12

– Residential Handler Exposure Scenarios for HHT

Scenario Exposure Route(s) Duration

Residential ExposuresDocket Number EPA

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P age 26 of 74 Scenario Exposure Route(s) Duration

Application of Paint by a Residential Painter (Airless Sprayer) Dermal Inhalation Short Term

Application of Paint by a Residential Painter (Brush/Roller) Dermal Inhalation Short Term

Application of Household Cleaners (Spray and Wipe) Dermal Inhalation Short Term

Application of Household Cleaners ( Mop ) Dermal Inhalation Short Term



### 3.3.4 Residential Post

- Application Exposures

During the registration review comment period, a registrant indicated their intent to support HHT residential post

- application uses such as treated paints, cleaners and detergents that were originally determined in the RED to be ineligible for reregistration

.

T he Agency intends to refine the inhalation risks to formaldehyde and HHT during registration review. The Agency anticipates the need to require inhalation air chamber tests for all of the household cleaner, detergent and paint uses to refine the residential HHT and formaldehyde vapor exposure risk assessment. The Agency anticipates the need to require th e s e data under the registration review data call

- in under the inhalation assessment guideline (Series 875.2500) and will conduct new residential post

- application risk assessments. The Agency anticipates the need to reevaluate the dermal and incidental ingestion risk assessments from t reated f loors and clothing. Table 13 lists the residential post

- application exposure scenarios that the Agency anticipates will be need ed for registration review.

Table 13

– Residential Post

- application Exposure Scenarios for HHT

Scenario Exposure Route(s) Duration

Residential Exposures Residential Bystander Exposures to Formaldehyde Emissions from Paint Inhalation Chronic
Residential Bystander Exposures to Formaldehyde Emissions from Household Cleaners Inhalation Chronic
Residential Bystander Exposures to Formaldehyde Emissions from Household Laundry Detergents Inhalation Chronic

Residential Exposure to Household Cleaners on Countertops and Floors Dermal Incidental Ingestion Short Term (Children)

Residential Exposure to Treated Clothes with Detergent Dermal I ncidental Ingestion Short TermDocket Number EPA

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## 3.4 Aggregate and Cumulative Exposure

3.4.1 Aggregate Exposures

The A gency anticipate s the need to conduct a n ew aggregate assessment during registration review to include the r esidential (paint, cleaners and detergents) and dietary uses

.


### 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 HHT and any other substances and HHT does not appear to produce a toxic metabolite produced by other substances. Therefore, f or the purposes of this registration review, EPA has not assumed that HHT 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/pesticides/cumulative/

. 

## 4 Environmental Risk Assessment

The Agency has not previously conducted a risk assessment that supports a complete endangered species determination for HHT

. The ecological risk assessment planned during registration review will allow the Agency to determine potential acute and chronic risks to aquatic organisms exposed to HHT transported from treatment sites into the aquatic environment. The risk assessment al so will allow the Agency to determine whether each use of the HHT 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

The Agency has received several studies regarding the hydrolysis of HHT including hydrolysis in buffered waters and metal working fluids, at different concentrations of HHT and different temperature regimes. HHT hydrolyzes rapidly at acidic pHs, shows slow hydrolysi s at more neutral pHs and is stable at basic pHs. In one study, at 25oC, HHT hydrolyzed within seconds at lower pHs (1 to 4), with a half

- life of 3.2 days at pH 5 , and with a half

- life of approximately 50 days at pH 7

. The rate of the hydrolysis reaction i s sensitive to temperature with more rapid breakdown seen at 37 o C than at 25 o C

. Hydrolysis at all pHs results in the release of f ormaldehyde

. After the release of formaldehyde from HHT, the remaining compound is 1,3,5

- trimethyl triazine

. A second h ydroly sis study designed to estimate the rate of formaldehyde release in aqueous m etalworking fluid was conducted at 25 o and 35 o C with HHT concentrations of 4, 5 and 6%

. Hydrolysis was observed, but approximately 30% of the material was unaccounted. Since t he study did not provide any rationale for this loss (MRID# 48741001) , it provides supplemental information on hydrolysis. The Agency anticipates requiring hydrolysis information on HHT which follows the formation and decline of the triazine degradate.Docket Number EPA

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P age 28 of 74 HHT i s not a volatile substance with a measured vapor pressure of 4.1 x 10

- 9 mm Hg at 25 o C

. Although a p ublished literature study (Voets, et al , 1975 ), concluded that HHT is a ready biodegradable pesticide , the study only measured the primary degradation of HHT and did not identify or follow biodegradation of the resulting compounds. In order to characterize the potential for biodegradation, additional information on the formation and decline of degradates is anticipated to be needed

.
The e stimated K oc of HHT i s 1 0 L/kg whi ch indicates it is mobile in soils. It has a low Log K ow of

- 2.3 and is therefore is not expected to bioaccumulat e in the food chain.

The Agency has a fairly complete environmental fate database for formaldehyde, but at this time, has no envi ronmental fate and transport data on the degradate 1,3,5

- trimethyl triazine and limited information on HHT

. It has no information on the down

- the

- drain data (ASRI, ASSI, Biodegradability) of HHT and 1,3,5

- trimethyl triazine. These studies are anticipated t o be required during registration review.



### 4.1.1 Water Quality

HHT is not identified as a cause of impairment for any water bodies listed as impaired under section 303(d) of the Clean Water Act

. 1 In addition, no Total Maximum Daily Loads (TMDL) h ave been developed for HHT

. 2 More information on impaired water bodies and TMDLs can be found at EPA’s website

. 3



## 4.2 Conceptual Model s for Environmental Exposure Pathways
Based on the summary of registered uses of HHT presented in Table 5 , physical/chemical properties and environmenta l fate data presented in Table 4 and Appendix B, the Agency ha s determined that some exposure of aquatic organisms to HHT and its degradates can potentially occur from releases of HHT and/or its degradates to surface water from industrial uses in metal working cutting fluids, and oil field drilling muds/workover flui ds/completion fluids; and in

service use of in

- can preservation of adhesives, chain lubricants, aqueous mineral slurries, and in

- service use of caulks, spackling, grout, adhesives, and foams.

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” (EPA

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- 0002) , found in the docket at www.regulations.gov , EPA

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. For industrial use of metal working cutting fluids, refer to slide 18. For industrial use of oil field drilling muds/workover fluids/completion fluids, refer to slides 19 and 20. For in

- service use of in

- can preservation of adhesives, chain lub ricants, aqueous mineral slurries, paints, stains, and coatings, refer to slides 24 and 25. For in

- service use of caulks, spackling, grout, adhesives, and foams, refer to slide 17. All relevant pathways of exposure will be considered when the Agency perfor ms a risk assessment for HHT and degradates.

1 http://ia spub.epa.gov/tmdl_waters10/attains_nation_cy.cause_detail_303d?p_cause_group_id=885

2 http://iaspub.epa.gov/t mdl_waters10/attains_nation.tmdl_pollutant_detail?p_pollutant_group_id=885&p_pollutant_ group_name=PESTICIDES
3 http://www.epa.gov/owow/tmdl/Docket Number EPA

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## 4.3 Ecological Effects Assessment

A hazard assessment was conducted for all uses of HHT i n 2008 (EPA, 2008). The hazard assessment was used to determine current labeling requirements for precautionary statements and to determine hazard endpoints for ecological organisms potentially exposed in the event of a spill or other unintended environmental releases. At that time, no ecological risk assessment was required for the registered uses. This was based on the use patt erns, which indicated that release and exposure levels were expected to be low when products were applied according to label directions and use precautions. The ecological risk assessment planned during registration review will allow the Agency to determin e potential acute and chronic risks to aquatic organisms exposed to HHT transported from antimicrobial use sites into the aquatic environment. Since HHT is a formaldehyde releaser, the Agency has reviewed the eco

- toxicity data for formaldehyde (EPA, 2008) and has compared the parent compound’s toxicity to that of formaldehyde. The toxicity patterns of the two compounds is similar, ranging from practically nontoxic to moderately toxic. Since formaldehyde data has been assessed in the RED and does not contrib ute new information to the HHT risk assessment, th ese data will not be included in this review

. Therefore, the risk assessment will be based on the HHT toxicity data.

4.3.1 Mechanism of Action HHT releases formaldehyde. T he hydrolysis of formaldehyde

- condensa tes at the cell boundary (wall) e ffectively denatures this peptidoglycan structure and, thus, leads to cellular death.



### 4.3.2 Measures of Effect (Ecotoxicology Endpoints) Ecological effects data are used as measures of direct and indirect effects to aquatic and terrestrial organisms. Acute and chronic toxicity data from registrant

- submitted studies conducted in accordance with the 850 OCSSP Harmonized Test Guidelines will be used to evaluate the potential direct and indirect effects of HHT to animals and plants. Additionally, as described below, relevant data from the EPA’s Office of Water and from the open literature also may be used to evaluate potential direct and indirect effects

. All available ecotoxicity endpoints from studies submitted by registrants are tabulated and referenced in the Ecological Effects Profile (Appendix C). OPP uses the most sensitive of these endpoints for assessing acute and chronic risks to each aquatic rece ptor group. The avian toxicity endpoint is used to determine the need for precautionary labeling for hazards to wildlife. The endpoints selected for the assessment for HHT from the data sub mitted to the Agency are provided in Table 14

. Data gaps exist for chronic freshwater fish, chronic freshwater invertebrates, algae, and aquatic vascular plants

, and these data are anticipated to be required

.

Table 14

– Ecological Effects Endpoints Selected f or Assessing Risks from HHT Use

Receptor Group Surrogate Species Risk Scenario Toxicity Endpoint Reference

Freshwater fish Bluegill Sunfish Acute 96

- h LC 50 = 39 mg/L MRID# 248883

Rainbow Trout Acute 96

- h LC 50 = 41.7 mg/L MRID# 247712
Freshwater fish Bluegill Sunfish Chronic Data gap 2

--

Daphnia magna Acute 48

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P age 30 of 74 1 Not required for this use pattern based on a lack of exposure to terrestrial organisms

. 2 Required for this use pattern.

3 There are no data available, but acute to chronic ratios using freshwater acute and chronic data and estuarine acute data can be used to estimate chronic toxicity to estuarine/marine organisms.

4 Not required for this use pattern base d on its mobility in the soil ( K oc = 10 L/kg) . HHT is not expected to bioaccumulate in the food chain (Log K ow =
- 2.3).



## 4.4 Exposure Analysis Plan

4.4.1 Aquatic and Terrestrial Wildlife Exposure Est imates

To perform an analysis of potential risks from exposure of aquatic animals and plants to HHT and/or its degradates as a result of industrial uses of HHT, t he Agency plans to use the General Population and Ecological Exposure from Industrial Releases module of E

- FAST (Exposure and Fate Assessment Screening Tool)

. The input data required to run this E

- FAST module include: (1) percent removal of antimicrobial during was tewater treatment; (2) environmental releases of antimicrobial to surface water in kg/site/day; (3) annual number of days of release to surface water; (4) Standard Industrial Classification (SIC) code(s) to represent the industrial uses being assessed; and (5) concentrations of concern ( COCs ) for aquatic animals and plants.

To perform an analysis of potential risks from exposure of aquatic animals and plants to HHT and/or its degradates from in

- service uses, t he Agency plans to use the Down

- the

- Drain module of E

- FAST

. The inputs required to run this module include percent removal of antimicrobial Freshwater invertebrates Chronic Data gap 2

-- Freshwater benthic invertebrates Freshwater midge Subchronic/ Chronic Not required 4

--

Estuarine/marine fish Sheepshead minnow Acute 96

- h LC 50 = >118 mg/L MRID# 43143102 Chronic Not required 3

--

Estuarine/marine invertebrates Mollusk Acute 48

- hr EC 50 = 2.3 mg/L MRID# 43175402

Shrimp Acute 96

- hr LC 50 = 12 mg/L MRID# 43143103

Chronic Not required 3

-- Estuarine/marine benthic invertebrates Marine amphipod Subchronic/ Chronic Not required 4

-- Algae Green alga Nonlisted Data gap 2

-- Aquatic vascular plants Duckweed Nonlisted Data gap 2

-- Seedling Emergence R ice Nonlisted Not required 1

-- Vegetative vigor Rice Nonlisted Not required 1

--

Birds Northern bobwhite

quail Acute LD 50 = 1520 mg/kg MRID# 43154301

Mallard duck Subacute

dietary LC 50 >5000 mg/kg MRID# 240113101

Chronic Not required 1

--

Mammals Acute LD 50 = 1250 mg/kg (males) LD 50 = 763 mg/kg (females) MRID# 41675206

Chronic Not required 1

-- Nontarget insects Honey bee Acute Not required 1

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P age 31 of 74 and/or its degradates during wastewater treatment, wastewater treatment plant influent volume, and COCs for aquatic animals and plants.

For both of these approaches , t he number of days of exceedance of Concentrations of Concern (COCs) for HHT and/or its degradates will be used to determine the magnitude of exposure to aquatic organisms. The method used to estimate environmental releases of the antimicrobial pesticide and/or its degradates to surface water will be based on the specific end

- use being evaluated

. Values for ecotoxicity endpoints will be used to develop COCs used in these E

- FAST modules.



### 4.4.2 Screening Level Down

- the

- Drain Analysis As stated in Appendix D, a screening

- level Down

- the

- Drain analysis of HHT was not performed since such an analysis is appropriate only for uses of antimicrobial pesticides that would be released solely to domestic wastewater treatment facilities. Uses of HHT include industrial uses that would be expected to have the potential to release HHT and/or its degradates to industrial wastewater treatment facilities and in

- service uses that would be expected to have the potential to release HHT and/or its degradates to domestic wastewater tr eatment facilities. Since HHT has the potential to be released to both domestic and industrial wastewater treatment facilities , results from a screening

- level down

- the

- drain analysis would provide incomplete information on the potential for risks to aquatic organisms from exposure to HHT and/or its degradates.



## 4.5 Effects Analysis Plan

Additional open literature studies will be identified through EPA’s ECOTOXicology (ECOTOX) database. The ECOTO X database will be searched during the risk assessment phase of HHT and the data reviewed following the Agency ’s policy for reviewing public literature for use in assessing risks from pesticide s

. 4 Those ecotoxicity values found in the public literature to be more sensitive than those currently in Table 14 and / or which satisfy an existing data gap(s) will be used during the risk assessment phase of HHT to update Table 14 . ECOTOX was created 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 Pro gram (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, sexu al 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 gr oups. As part of its reregistration decision for HHT , EPA reviewed these data and

4 http://www.epa.gov/pesticides/science/efed/policy_guidance/team_authors/endangered_species_reregistration_wor kgroup/esa_evaluation_open_literature.htmDocket Number EPA

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P age 32 of 74 selected the most sensitive endpoints for relevant risk assessment scenarios from the existing hazard database. However, as required by FFDCA section 408(p), HHT is subject t o 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 a n 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 screenin g 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 hormona l 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 ident ified for EDSP screening was published on June 14, 2013 5 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. 6


## 6 Label Changes

T he Agency is taking steps to bring outstanding HHT la bels into compliance with risk mitig ation measures from the HHT RED. A pplication rates should be lowered for certain HHT uses, including for oil field and metalworking fluids uses . In addition, all paints, stains, and coatings

uses , and institutional and h ousehold cleaning product uses need to be removed to address RED mitigation. The Agency will continue to pursue label compliance through regulatory or other action during registration review . Until these uses or rates are changed, the Agency will anticipate the need to require additional data and will assess these uses and higher use rates during registration review. In addition, t he Agency invites submission of 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.

5 See http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2009

- 0477

- 0074 for the final second list of chemicals.

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## 7 Next Steps

A DCI will be developed requiring generation and submission of the data listed under the “Risk Assessments and Anticipated Data Needs” section of this document. The Agency expects to issue the DCI by June of 2016.Docket Number EPA

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## 8 References

MRID 47284307. Hutmacher, H. (2002) Vapour Pressure of Myacide HT T: Final Report. Project Number : 01L00575. Unpublished study prepared by BASF Aktiengesellschaft, Labor fuer Oekotoxicologie. 12 p.

MRID 47284308. Dogemuller, A. (2002) Partition Coefficient n

- Octanol/Water [log P(ow)] of Myacide HT T: Final Report. Project Number: 01L00533. Unpublishe d study prepared by BASF Aktiengesellschaft, Labor fuer Oekotoxicologie. 15 p.

MRID 473354

- 02. A Study of Formaldehyde Exposures from Metal Working Fluid Operations Using Hexahydro

- 1,3,5

- Tris (2

- Hydroxyethyl)

- S

- Triazine. H.J. Cohen, University of New H aven, May 3, 1994.

Holtzman, G.H., and Rossmoore, H.W

. 1977

. Evaluation of Action of a Formaldehyde Condensate Germicide

. Dev Ind Microbiol 18:753

- 758.
Linnainmaa, 2003. “Control of Worker’s Exposure to Airborne Endotoxins During Use of Metalworking Fluid s,” Markuu Linnainmaa, Hannu Kivirata, Juha Laitinen and Sirpa Laitinen, Kuopio Regional Institute of Occupational Health and National Public Health Institute Department of Environmental Health, Kupio, Finland, Published in the AIHA Journal 64:496

- 5000, Ju ly/August 2003.

Kline, 200 6 . Specialty Biocide s 2004

- 2005, Volume II: United States , Kline and Company, Inc.

U.S. EPA, 2002. ECOTOX User Guide: ECOTOXicology Database System. Version 3.0. Available: http://www.epa.gov/ecotox/

U.S. EPA, 2008. Ecological Hazard and Environmental Risk Assessment Chapter: Formaldehyde, Paraformaldehyde. January 29, 2008

.
U.S. EPA, 2008

. Reregistration Eligibility Decision (RED) for Grotan (HHT)

. June, 27, 2008.

Voets, JP, Pipyn, P, Van Lanker, P, and Verstraete, W: 1975: De gradation of Microbiocides under Different Environmental Conditions, J. Applied Bact.: 40:67

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# Appendix A T oxicology Profile

Acute Toxicity for Product Labeling

As listed in Table A1 , HHT was found to be low toxic for both acute oral and dermal toxicity (Toxicity Category III for both) . An acute inhalation toxicity study is not available in the Agency ’s hazard database and is required. This study was identified as a data gap in the RED. HHT wa s shown to be corrosive to the eyes ( Toxicity Category I ) but mildly irritating to skin (Toxicity Category IV)

. HHT wa s shown not to be a dermal sensitizer.

Table A 1. Acute Toxicity Studies for HHT

Guideline No./ Study Type MRID No. Results Toxicity Category 870.1100 Acute oral toxicity

– Rat (Purity 79.4%) 41675206 LD 50 = 1250 mg/kg (males); LD 50 = 763 mg/kg (females) III

870.1100 Acute oral toxicity

– Mouse (Purity 78.5%) 00155959 LD 50 = 1.30 (1.14

– 1.48) ml/kg III

870.1200 Acute Dermal Toxicity

– Rabbit ( Purity 79.96%) 00155984 LD 50 > 2000 mg/kg III

870.1300 Acute Inhalation Toxicity

– Rat N/A N/A N/A

870.2400 Primary Eye Irritation

– Rabbit ( Purity 79.96%) 00155985 Corrosive I

870.2500 Primary Dermal Irritation

– Rabbit ( Purity 79.96%) 00155986 Mild irritant IV

870.2600 Dermal Sensitization

– Guinea

Pigs ( Purity 79.96%) 00155987 Not a sensitizer N/A

N/A=Not available 

## Subchronic Toxicity

Adequacy of Database for Subchronic Toxicity: The database for subchronic toxicity is considered incomplete. There are several studies in the Agency ’s hazard database to address subchronic oral toxicity , including a 90

- day oral (gavage) toxicity study (MRID 41 4830 01) , a 90

- day oral (drinking water) toxicity study (MRID 47300901), a 21

- day (MRID 00155989) dermal study, and two 90

- day dermal toxicity st ud ies (MRID s 41 4830 02 & 41858301 ) ; all theDocket Number EPA

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. A brief summary of each of the studies is provided below. A subchronic oral toxicity in non

- rodent species and a route

- specific subchronic inhalation toxicity study in

r oden ts are anticipat ed to be required for registration review risk assessment

, based on the currently registered uses for HHT and its high volatility; these studies were identified as data gaps in the RED and are part of the data required by the post

- RED DCI issued on May 12, 2010

.
870.3100 90

- Day (13

- Week) Oral Toxicity

– Rat

(1) In a subchronic oral toxicity study (MRID 41483001), groups of Crl:CD(SD)BR rats (10/sex/group) received the test material HHT (78.5% a.i.) via gavage at doses of 0, 10, 50, 100, or 250 mg/kg/day, 5 days/week for 13 weeks

. Mortalities were reported during the study period

. With the exception of one female from the high dose group, the decedents had gross changes in the lung and/ or thoracic cavity

. The lungs in these animals did not collapse upon opening the thoracic cavity and adhesions or fluid was present in the area

. Inflammatory changes and vascular congestions were present in the thoracic viscera

. In addition to the thoracic lesions, all decedents from the high dose group showed a white discoloration and prominence of the limiting ridge of the stomach.

There were significant differences in the body weight gain and food consumption between treated and control rats

. A bsolute ad renal weights were increased for females in the high

- dose
group , although i t was not statistically significant ; a n increase in splenic weight was also observed in all treated males

. The sponsor attributes this difference to unusually low weights in control animals.

Grossly, a white discoloration and/or prominence of the limiting ridge of the stomach were present in all high

- dose males and in the remaining high dose females which survived the duration of the study as well as in some of the animals in the sec ond high dose groups

. This gross finding was correlated with the treatment

- related microscopic lesions in the stomach , including erosion of the glandular mucosa of the stomach, e pithelial hyperplasia , and l ymphocytic infiltration

. Microscopic lesion i n the liver was also observed including increased incidence of hepatocyte vacuolation in high dose males.

A statistically significant reduction in hematocrit w as found in all treated males

. Additionally, males in second high

- dose group demonstrated a statistica lly

- significant decrease in hemoglobin concentration

. Although these findings were statistically significant, they were within the expected laboratory range.

Significant differences in blood chemistry values included reduced bilirubin in all treated groups

, reduced total protein in both sexes of rats in high

- dose group , and a statistically significant increase in sodium concentration for all treated males and for females in all treated groups except for the lowest dose group

. The bilirubin and total protein values were within the normal range and were not considered to be biologically significant

. A high frequency of peribronchiolar lymphoid hyperplasia was reported for all groups

. The incidence of this finding ranged from 70 to 100 percent on both sexes

. W hile this finding is notDocket Number EPA

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P age 37 of 74 believed to be treatment

- related, the significance of this finding was not addressed in the study report

. Based on the results of this study, the s ystemic NOAEL is 50 mg/kg/day, and the systemic LOAEL is 100 mg/kg/day, based on gas tric lesions (erosion of the mucosa and lymphocytic infiltration) in males and females.

This study is classified as Minimum

. (2) In a 90

- day oral toxicity study (MRID 47300901 )

, Hexahydro

- 1,3,5

- t ris

- (2

- h ydroxyethyl)

- S

t riazine (79% a.i. , Batch # 1157) was administered to 10 Wistar rats/sex/dose in water at dose levels of 0, 200

, 1000 , or 5000 ppm (equivalent to 0, 14.1

, 64.1

, or 285.2 mg/kg bw/day for males and 0, 21.2, 91.0, or 338.6 mg/kg bw/day for females ).

High

- dose animals showed reduced food and wat er consumption, reduced body weight (males only), reduced food efficiency, increased blood urea concentrations, and increased urine specific gravity as compared to the control group

. Decreased water consumption was also observed in mid

- dose (1000 ppm) anim als but was not considered an adverse effect, as urinary and blood chemistry did not suggest dehydration

. The mean absolute weight of the kidneys was significantly increased in the high

- dose group, while the relative weight of the kidneys was significantly increased in females of both the mid

- and high

- dose groups

. However, this change did not correlate to gross or microscopic changes in the kidney and therefore is not considered an adverse effect

. The mean relative weights of the brain and adrenal glands w ere significantly increased in high

- dose males, which are attributed to the weight loss observed in these animals.

The neurological observations revealed no signs of a dose

- dependent relationship

. However, no statistical tests were performed on the neurolo gical data

. Other clinical signs, such as alopecia and an injury of the right forelimb, were observed in the high

- dose group, but were isolated, spontaneous in nature, and reflected the natural range of variation.

The LOAEL is 5,000 ppm ( 285.2 mg/kg body w eight/day in males; 338.6 mg/kg body weight/day in females) , based on decreased body weight and body weight gain, food and water consumption, and changes in clinical urinary and blood chemistry

. The NOAEL is 1,000 ppm (64.1 mg/kg body weight/day in males; 91.0 mg/kg body weight/day in females)

.
This 90

- day oral toxicity study in the rats is A cceptable /G uideline and satisfies the guideline

r equirement for a 90

- day oral toxicity study (OPPTS 870.3100; OECD 408) in rats

.
870.3200 Subchronic (21

- day dermal) Toxicity

– Rat

In a subchronic dermal study ( MRID 00155989), Glokill 77 (78.5 % a.i., batch TS

- 704) was administered dermally to groups of Sprague

- Dawley rats (5/sex/dose) at concentrations of 0, 100, 500 or 1000 mg/kg/day for 21 days

. The test material was used undiluted as supplied

. One group served as controls and was treated with sterile distilled water in similar fashion as the test compound

- treated animals

. The test compound was held in contact with the skin by a porous
gauz e dressing covered with an Elastoplast elastic adhesive bandage

. At the end of each 6

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P age 38 of 74 All animals survived the study

. No toxic signs were reported except for local dermal irritation, which was dose

- related and increased in severity with the duration of the study

. All animals gained weight during the study; however, all groups of treated males gained less weight than corresponding controls

. Male weight gain in all groups was approximately 70% of the control male weight gain

. The weights observed for all three treatment groups was within the range normally observed in the investigators’ laboratory, and therefore were not considered treatment related

. Weight gain was comparable in control and treated females.

No difference in food consumption was reported between control and treated males, but during week 1, the mid and high

- dose female food consumption was significantly decreased (p < 0.05) when compared to the co ntrols

. No intergroup difference in water consumption was reported.

Mean corpuscular volume was reduced in the low and high

- dose males, and hemoglobin was reduced in the low and high

- dose females, but the reductions were not considered biologically signifi cant since they were within the normal range of values for Sprague

- Dawley rats in the investigators’ laboratory.

The following statistically significant changes were noted in the treated animals when compared to the controls (p <0.05) , including increased protein in mid and high dose males and high dose females, increased sodium in all treated males, increased phosphate in mid and high dose females, increased AST in mid and high dose males, decreased A/G ratio in mid and high dose males and high dose female s, and decreased chloride in mid and high dose females

. These changes were considered biologically insignificant because the values observed were within the range observed in the animals used in the hematology and biochemical screens

. Except for multifocal scab formation at treatment sites at all dose levels, no macroscopic abnormalities were reported

. Absolute adrenal weight was significantly decreased in low and high

- dose males and was reported to be due to two control animals with high adrenal weights

. No difference was reported in relative organ weights

. Treatment

- related histopathological changes were reported at treated skin sites, but in no other tissues examined

. Histopathological findings included dose

- related epidermal ulceration, acantho sis and dermal inflammatory infiltration with occasional necrosis and hemorrhage in the treated animals.

Based on the dose related epidermal ulceration and dermal inflammation reported at the treated skin sites , the dermal LOAEL i s 100 mg/kg /day , and the dermal NO A EL i s not obtained in this study

. The systemic NO A EL is 1000 mg/kg /day , the highest dose tested ( HDT ), and no LOAEL for systemic toxicity was established

. This study is classified Minimum

.
870.3250 Subchronic (90

- day dermal) Toxicity

– Rat

(1) In a sub chronic dermal study (MRID 4185 8 3 01), Vanicide TH (96.6% a.i.) was administered dermally to groups of Crl:CD BR (Sprague Dawley) rats at concentrations of 0, 10, 30 or 100 mg/kg/day

. A n area from the shoulders to the lumbar region was clipped and div ided into quadrants

. Daily applications were rotated from quadrant to quadrant

. After the material was applied with a gauze patch, the patch was wrapped with COBAN

. Material was left in contactDocket Number EPA

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P age 39 of 74 with the skin for 6 hours and afterward, the residual material was removed with a paper towel and was washed with reverse osmosis water

. This procedure was repeated for 5 days a week for the next 13 weeks

. The test material was applied to approximately 10% of the body surface area in one of the prepared quadrants.

Th ree females died during the study (1 control, 1 mid and 1 high dose)

. The death in the control animal was caused by renal and urinary bladder problems and were believed to be associated with dosing procedures in the treatment groups

. In addition, all high dose animals were sacrificed

o n Study D ay 36 because of the severity of the dermal irritation , including erythema, eschar, desquamation, necrosis, exfoliation and ulceration

. Females in the high dose group were reported to have the most severe dermal lesio ns , and no healing was observed after the weekend periods when the test material was not administered

. S ome healing was present after the weekends i n animals in groups 2 and 3

. S evere erythema with eschar

, as well as e xfoliation, necrosis, desquamation and ulceration

, were reported in the mid dose animals , while slight erythema with eschar was observed for low

- dose animals

. A statistically significant decrease (7%) in body weight was reported for males in the high dose group when compared to controls

. Alth ough there was no statistical significance in mean body weights in the low and mid dose groups, there appeared to be a linear response to the dose of the test material when these groups were compared to controls

. No differences in mean body weights

were re ported for females in any of the treated groups when compared to controls.

In the mid and high dose males, mean weekly food consumption was statistically lower than controls on day 14

. This was the only reported difference in food consumption reported for males

. In females, a statistical significance was reported for animals in the mid and high dose groups on day 35; however, the significance was the result of an increase in food consumption reported for these animals.

I n the interim evaluation

, s tatistically significant differences in hematology were reported in both sexes in the high dose group , including lower hemoglobin

, hematocrit , and r ed blood cell counts

, when compared to controls ; platelet counts were higher

. In high

- dose females, white bl ood cell counts were also higher than controls during the interim evaluation

. At the end of the study, no significant differences were reported for any of the hematology parameters

. In all treated males, albumen, calcium, total protein, sodium and chlorid e values were statistically lower than controls at the interim evaluation

. Phosphorus values were statistically decreased in mid

- dose males

. Potassium, AST and ALT values were lower than controls in high

dose group animals

. In females, significant differen ces reported for albumen, BUN calcium, glucose, total protein, sodium, chloride and potassium during the interim

. Most of these values were higher than controls with the exception of albumen and total protein in high dose animals

. Terminal serum chemistry values revealed lower albumen, sodium, calcium and total protein values in low

- and mid

- dose males

. Low

- dose males also had lower chloride and potassium levels when compared to controls

. In females , a few clinical chemistry alterations were reported, inclu ding increased phosphorus and decreased bilirubin and cholesterol in mid

- dose group, when compared to controls

. D ecreases in total protein and cholesterol were reported in low

- dose females

.Docket Number EPA

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P age 40 of 74 No significant differences were reported for organ weights

. Relat ive increases were reported for liver and lung weights in females without statistical significance when compared to controls.

Histologically, there were no treatment related lesions reported with the exception of those reported for the skin

. T here was an i ncreased incidence in hyperplasia and hyperkeratosis of the epidermis of the skin in both sexes

. Sebaceous gland hyperplasia and denial inflammatory cell infiltrations were also increased; other lesions included eschar formation, focal dermal fibrosis, foc al epidermal necrosis and focal sebaceous gland necrosis.

T he d ermal NO AEL i s < 10 mg/kg /day , the lowest dose tested (LDT); the dermal LOAEL i s 10 mg/kg/day , based on dermal irritation and histological dermal lesions characterized by hyperplasia, hyperkeratosis, inflammation, sebaceous gland hyperplasia and eschar formation.

Observations made with regard to serum chemistry and hematology values in both sexes appear to be incidental findings that are not biologically significant

. No significant diff erences were reported for the hematology values at the end of the study and the serum chemistry values that were statistically significant at both the interim and terminal evaluations, were within the range of normal values

. The s ystemic NO A EL was 30 mg/kg and the systemic LO A L was 100 mg/kg

, based on severe dermal irritation in both sexes and decreased body weight reported in high dose males

. At the highest dose tested, both sexes of animals had to be sacrificed on day 36 due to the severity of the dermal lesions.

This study is classified Minimum

. (2) In a subchronic dermal study (MRID 41483002), groups of Crl:CD(SD)BR rats (10/sex/group) received the test material HHT (78 .5 % a.i.) to the clipped dorsal area at doses of 0, 5, 50, or 250 mg/kg/day 6 hours/da y, 5 days/week for 13 weeks

. During the exposure period, the application sites were covered with semi

- occlusive bandages and animals were fitted with Elizabethan collars to minimize ingestion of the test chemical

. The test site was not washed between doses

. No deaths were reported during the study

. The clinical signs observed during the study included periorbital, perianal, and cranial fur staining in all groups; yellow staining at the applications site in animals from low

- and mid

- dose groups

. S kin reacti ons at the applications site were observed in animals from mid

- and high

- dose groups , including erythema scored as very slight to severe with increase in incidence and severity in high

- dose animals

. Additionally, e dema was present and characterized as bein g very severe to moderate in high

- dose animals ; whereas, only isolated incidences of edema were reported in mid

- dose group

. Occasional incidences of erythema were observed in female s in the low

- dose group

. Statistically significant differences were found i n blood chemistry values from individual animals , including increased alanine aminotransferase ( one low

- dose male ), increased phosphorous ( one low

- dose female ) and increased chloride ( one low

- dose female )

. In mid

- dose
group, blood chemistry parameters were increased in two animals , o ne female had increased sodium and one male had increased chlorine levels

. In high

- dose group , one female had elevated glucose levels and one male had elevated sodium levels.Docket Number EPA

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P age 41 of 74 Microscopically, there was an increase in the inciden ce of epidermal hyperplasia and ulceration of the treated area of the skin in animals of the mid

- and high

- dose groups

. Eighty to 100 percent of both the treated and control animals were found to have peribronchiolar lymphoid hyperplasia in the lungs

. The significance of the lesions was not discussed.

The dermal NOAEL i s 5 mg/kg/day and the LOAEL i s 50 mg/kg/day, based on skin lesions observed at this level

. The systemic NOAEL i s greater than 250 mg/kg/day , highest dose tested (HDT) and the systemic LOAEL is not established

. This study is classified as Minimum

. 

## Developmental Toxicity

Adequacy of Database for Developmental Toxicity: The database for developmental toxicity is considered incomplete. There are two studies in the Agency ’s hazard database to address developmental toxicity ; both studies are in the rat (MRID s 41161801 & 41865701 ). A brief summary of each of the studies is provided below. A developmental toxicity study in non

- rodent species is anticipated to be required for registrat ion review risk assessment

, based on the currently registered uses for HHT; this study was identified as a data gap in the RED and is part of the data required by the post

- RED DCI issued on May 12, 2010

.
870.3700a Prenatal Developmental Toxicity (Gavage) Study

– Rat

(1) In a developmental toxicity study in rats (MRID 41161801), artificially inseminated female Sprague

- Dawley rats (24/group) were administered HHT (78.5%) by gavage at doses of 0, 250, 500, and 750 mg/kg/day in deionized water on gestation days 6 through 15

. One high

- dose dam was sacrificed on GD 14. Prior to sacrifice on that day the animal exhibited labored breathing, rales, emaciation, piloerect ion, red staining around the mouth and nose and pale extremities. All other animals survived the study.

High dose females exhibited post dosing salivation

. Rales, labored breathing, wheezing, and tachypnea were observed occasionally in the mid and high do se groups toward the end of the dosing period. No other clinical signs were reported.

Maternal body weight gain (p<0.01) and food consumption (p<0.001) was significantly lower in the high dose females during the dosing period than the controls

.
Stomach le sions, characterized by ulceration and/or scarring of the mucosa were observed in 14 of 20 high dose females, including the dam sacrificed on day 14. No gross abnormalities were reported in the other dosage groups.

The data did not demonstrate any differen ces between the control and treated dams with respect
to pregnancy rates, number of corpora lutea, implantation sites, number of live fetuses, or early and late res orptions . There were no abortions and no premature deliveries. In addition, those dosage lev els did not produce any developmental toxicity as measured by fetal pup weight,
externa l, or visceral, abnormalities. There were increased incidences of vestigial 14 th ribs andDocket Number EPA

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P age 42 of 74 retarded ossification of the vertebral thoracic centra whic h appeared to be dos e

- related

. However, since they were not statistically significant, and the incidence of these abnormalities is highly variable in rats, they are not considered treatment

- related.

The m aternal NOAEL is 500 mg/kg/day and the LOAEL is 750 mg/kg/day, based on decreased body weight gain, ulcerations and/or scarring of the stomach mucosa. The Developmental toxicity NOAEL is greater than or equal to 750 mg/kg/day , the highest dose tested (HDT)

. This study is classified as Minimum

. (2) In a developmental t oxicity study in rats (MRID 41865701), Vanicide TH (96.6% a.i.) was administered via gavage to groups of female Crl:CD BR Sprague

- Dawley rats (25/dose) at concentrations of 0, 10, 75 or 150 mg/kg/day from day 6 through 15 of gestation

. Mortality was obser ved in the control (1/25), mid

- dose (2/25) and high

- dose (6/25) groups but these were thought to be associated with incorrect gavaging technique.

Body weight gain was significantly reduced in the high

- dose group (39%) when compared with the control animal s

. Food consumption was higher for the low

- dose and mid

- dose groups through the dosing period and for the low

- dose group during the post dosing period

.
No treatment related lesions were observed in the maternal animals at sacrifice

. Lesions observed in th e cervical lymph nodes, liver, lung, kidney, skin and fur were reported as either single observances or at incidences that would not suggest an association with the compound.

Post implantation losses were higher in the treatment groups as compared with th e controls but only the low

- dose group was statistically significant.

No differences were reported for fetal body weights when control groups were compared with the treatment animals

. External variations reported included protruding tongue; swollen hind
p aws with hematoma and stunted fetuses but there was no statistical significance in the treatment groups for the above mentioned observations

. Malformations such as encephaly, micrognathia, omphalocele, and thread like tail were observed in the control, low

- dose and mid

- dose groups. No such malformations were seen in the high

- dose group

. An increase in the incidence of hypoplastic sternebrae was reported for animals in the low

- and mid

- dose groups

. There was a statistically significant increase in both the f etal and litter incidence of this anomaly

. The fetal incidence of unossified sternebrae was statistically increased in the mid

- dose group when compared to controls but this could be an incidental finding

. Visceral variations were observed for all groups wi th dilated renal pelvises and convoluted and/or distended ureters being the most common variations observed

. There was a statistically significant increase in this observation in the high

- dose pups and a treatment related trend was observed for the occurre nce of dilated renal pelvises and for bilateral convoluted ureters.

Based on the results of this study, the maternal NOAEL was 75 mg/kg/day and LOAEL was 150 mg/kg/day , based on decreased body weight gain. The developmental NOAEL was <10Docket Number EPA

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P age 43 of 74 mg/kg/day

, the lo west dose tested (LDT), based on the observed significant positive trend in the incidence of bilateral convoluted ureters and dilated renal pelvises.

This study is classified Supplementary

. However, historical control data should have been provided to det ermine whether the renal observations in the pups were treatment

- related. 

## Reproductive Toxicity

Adequacy of Database for Reproductive Toxicity: The database for reproductive toxicity is considered incomplete

. There are no reproductive toxicity studies in the Agency ’s hazard database; such a study is anticipated to be required for registration review risk assessment

, based on the currently registered uses for HHT

. This study was identified as a data gap in the R ED and is part of the data required by the post

- RED DCI issued on May 12, 2010

. 

## Chronic Toxicity

Adequacy of Database for Chronic Toxicity: The database for chronic toxicity is considered incomplete

. There are no chronic toxicity studies in the Agency ’s hazard database; these studies are anticipated to be required for registration review risk assessment

, based on the currently registered uses for HHT

. The chronic studies were identified as data gaps in the RED and are part of the data required by the post

- RED DCI issued on May 12, 2010

. 

## Carcinogenicity

Adequacy of Database for Carcinogenicity : The database for carcinogenicity is considered incomplete

. There are no carcinogenicity studies in the Agency ’s hazard database; these studies are anticipated to b e required for registration review risk assessment

, based on the currently registered uses for HHT

. The carcinogenicity study were identified as data gaps in the RED and

are part of the data required by the post

- RED DCI issued on May 12, 2010

. 

## Mutagenicity

Adequacy of D atabase for Mutagenicity : The database for mutagenicity is considered adequate for the currently registered uses

. Several mutagenicity studies have been submitted and reviewed by the Office of Pesticide Programs (OPP) ; some of the in vitro studies were not acceptable

. HHT is found to be mutagenic in a few in vitro studies ( e.g. , chromosome aberration assay and UDS assay ) but not in the corresponding in vivo studies

. The summaries of these studies are provided below.

Gene Mutation

(1) In a bacterial reve rse mutation assay (MRID 41231702 ), following a range

- finding toxicity testing with TA98 (only) at doses up to 5000 μg/plate, triplicate cultures of all five Ames strains (TA1535, TA1537, TA1538, TA98, TA100) were exposed to Triazine (78.5%) for 72 hours (extended from the customary 48 hours because triazine is bacteriostatic, thus slowing growth of revertent colonies) at each of five dose levels (0.32, 1.6, 8, 40, and 200 μg/ plate), both in theDocket Number EPA

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P age 44 of 74 absence and presence of a mammalian me tabolic activation system consisting of the liver microsomal fraction (S9) prepared from male Fischer 344 rats induced by Aroclor 1254 and containing 44 mg protein per mL, plus generating cofactors

. The entire experiment was repeated.

In both experiments, the test substance consistently reduced background lawns at the HOT, 200 μg/mL, in the absence of activation but with one exception did not induce significantly increased revertent colony counts

. The exception was a significant F

- value (p < 0.05) calculate d for inactivated TA1537 cultures, derived from a wider

- than

- usual variation in mean numbers of revertents on test plates (3.3 to 7.7) compared to the concurrent mean control value (4.7)

. This was discounted by the investigator because there was neither a doubling of revertent rate nor evidence of a dose

- relationship

. Hence, the investigator concluded that triazine was not mutagenic in the Ames testing at levels up to levels of toxicity.

(2) In a reverse gene mutation assay in bacteria (MRID 47284305) , stra ins TA1535, TA100, TA102, TA 1537, TA 98 of S. typhimurium and E. c oli WP2 urvA were exposed to Protectol HP (purity not provided, Batch No.: PK95/222) in water at the following concentrations: 0, 100, 500, 2500, 5000, 7500  g/plate in the presence and absence of mammalian metabolic activation using the plate

- incorporation method for strains TA1535, TA 100, TA1537, and TA98 ; and 0, 25, 50, 100, 200, and 400  g/plate in the presence and absence of mammalian metabolic activation using the plate

- incorporatio n and preincubation method for strains TA1535, TA 100, TA1537, TA98

, TA102 and E.Coli WP2 uvrA.
Protectol HP didn’t induce any dose

- dependent increases in mean revertants/plate in any strains over all concentrations and conditions

. The positive controls ind uced the appropriate responses in the corresponding strains

. There was no evidence of induced mutant colonies over background

. There was inconsistency in the results reported for the TA102 plate incorporation test; TA102 +S9 did not have a clear positive c ontrol response, thus, a new test was conducted that elicited an appropriate increase in revertant colonies

. This study is classified as Una cceptable /Non

- g uideline (upgradable) due to the lack of detailed description of the test material (such as no purity information provided).

(3) In a mammalian cell gene mutation assay at the HPRT locus (MRID 47284303 )

, Chinese Hamster V79 cells cultured in vitro were exposed to Protectol HT (79% aqueous solution , batch # 1157) in deionized water at concentrations of 1.6, 3.1, 6.3, 12.5, 25, or 30 μg/mL,

- S9 and at concentrations of 1.6, 3.1, 6.3, 12.5, 20, or 25 μg/mL, + S9 for a period of 4 hours.

Cultures exposed to the test material did not induce dose

- dependent increases in the number of
mutan t colonies per 10 6 cells and the mutant frequencies remained within the range of historical negative and solvent control values

. The positive controls induced the appropriate response. Mutation frequencies three times above the historical/corresponding neg ative and/or solvent controls, a criterion for a positive response, were not observed in cell cultures treated with Protectol HT at any concentration

. Therefore, Protectol HT was considered non

- mutagenic under the conditions of this assay.

This study is cl assified as A cceptable /G uideline and satisfies the guideline requirement for Test Guideline OPPTS 870.5300, OECD 476 for in vitro mutagenicity (mammalian forward gene mutation) data

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P age 45 of 74 Cytogenetics

(1) In a n in vitro mammalian cell cytogenetics chromosome aberration assay (MRID 47284302), Chinese H amster V79 C ells (CHC) were exposed to Protectol HT (79 g per 100g aqueous solution, Batch No. Partie

- Nummer 1157) dissolved in deionized water (10% v/v) at concentrations of 0, 2.5, 5, 10, 20, 30, or 40 μg/mL with and without metabolic activation (S9

- mix) for 4 hours.

Protectol HT was tested up to cytotoxic concentrations, as determined by an initial range

- finding study . The range

- finding study indicated that the test substa nce had clastogenic effects after 4 hours ; therefore, a second experiment with a longer exposure time was not performed. At concentrations of 20, 30, and 40 μg/mL the number of cells was reduced to 58, 56, and 33% of the concur rent solvent controls

– S9; an d 61, 45, and 41% of solvent controls +S9, respectively. At a concentration o f 30 μg/mL

- S9, the number of cells in metaphase was decreased to 45% of the control value. Greater test concentrations (30 μg/mL and 40 μg/mL +S9 and 40 μg/mL

- S9) were not evalua ted for mitotic indices, chromosomal aberrations, or polyploidy since the test substance was highly toxic at these levels, as shown by the decreased number of metaphase cells.

Dose

- dependent increases in aberrant cells, excluding gaps, occurred at doses of 10, 20, and 30 μg/mL

- S9 and at a concentration of 20μg/mL, +S9. A biologically relevant increase in the number of cells exhibiting exchanges occurred at concentrations of 10, 20, and 30 μg/mL

– S9 and 20 μg/mL +S9, as compared to solvent controls. No dose

- related increases in the rate of polyploidy were found after test substance treatment at any concentration ± S9. Positive controls induced the appropriate response. There was evidence of chromosome aberrations induced by the test material over background incidence rates

. Protectol HT is considered to be clastogenic in in vitro chromosomal aberration assay in Chinese H amster V79 C ells in the absence as well as presence of S9 activation.

This study is classified as A cceptable/Guideline and satisfies the guid eline requirement for Test Guideline i n vitro mammalian cytogenetics chromosome aberration OPPTS 870.5375; OECD 47 in cultured Chinese hamster V79 in vitro cytogenetic mutagenicity data

. (2) In an NMRI mouse bone marrow micronucleus assay (MRID 47284301), 6 males/dose were treated once, intraperitoneally, with Protectol HT (79% a.i., Partie

- Nummer 1157) at doses of 0, 37.5, 75, or 150 mg/kg body weight

. Bone marrow cells were harvested at 24 hours post

treatment for all dose levels, and 48 hours post

- treat ment for the negative control and 150 mg/kg treatment dose

. The vehicle was deionized water

. The dose range used for the micronuclei was informed by a preliminary study in which unscheduled deaths were noted in male and female mice at doses ≥ 200 mg/kg

. In the main study, there were signs of toxicity in treated animals (i.e., reduction of spontaneous activity, eyelid closure, apathy, abdominal position

, and death)

. A slight increase in micronuclei was noted in the high

- dose animals, indicating cytotoxic e ffects of the test substance

. This increase

, however, was not higher than the range of negative control values and was not found to be statistically significant using the Mann

- Whitney test

. Protectol HT was tested at an adequate maximum dose based on clini cal signs of toxicity, lack of unscheduled deaths, and evidence of bone marrow cytoxicity

. The positive control induced the appropriate response

. A significantDocket Number EPA

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P age 46 of 74 increase in the frequency of micronucleated polychromatic erythrocytes in bone marrow was not ob served at either analysis time point.

This study is classified as Acceptable/Guideline and satisfies the guideline requirement for Test Guideline OPPTS 870.5395; OECD 474 for in vivo cytogenetic mutagenicity data

. (3) In a mammalian bone marrow chromosome aberration test in mouse (MRID 41231701), Triazine (78.5%) was administered to groups of CD

- 1 mouse (5/sex/group) by gavage in a single dose at 200, 400, and 800 mg/kg, following a rang

- finding study (at doses up to 2000 mg/kg) and

were sacrificed 24, 48, or 72 hours later

. Control groups received 0.9% saline (solvent control), or cyclophosphamide (CPA, positive control) ; the latter sacrificed only at 24 hours

. In the range

- find ing test, deaths occurred at 1200 and 2000 mg/kg (respectively, 2 of 3 animals and 3 of 3 of each sex), but no mortalities at 800 mg/kg or below, providing an estimated LD 50 of approximately 1000 mg/kg

. Based upon the (OECD) convention of applying 80 % of the LD 50 as the MTD for this type of study, the HDT for the main study was sele cted as 800 mg/kg, with two lower doses of 400 and 200 mg/kg.

In the micronucleus assay itself, two high

- dose (800 mg/kg) animals (one of each sex) died before their scheduled sacrifice at 48 hours

. No cytotoxic i ty, as measured by reduction in PCE/NEC ratios, was found in any nabam

- treated group

. Random variation around control values for numbers of micronuclei was found at all sacrifice times and in both sexes of test animals

. A singular statistically significant increase in micronuclei over concurrent control was calculated for the 400 mg/kg male group sacrificed at 72 hours (0.14 vs. 0.02)

. This increase was considered to be of no biological significance since it fell within solvent control ranges at other sampling times (notably 24 hours when optimal ly most of the micronuclei would be visualized).

Hence, the investigator concluded that triazine did not induct micronuclei in bone marrow cells of mice treated orally up to a nominal dose (800 mg/kg) considered a MTD by OECD (80% of the LD 50 ).

Other Geno toxicity

(1) In a primary rat hepatocyte unscheduled DNA synthesis (UDS) assay (MRID 41262301 ), hexahydro

- l, 3, 5

- tris(2

- hydroxyethyl)

- s

- triazine was tested at nine dose levels ranging from 0.0001 to 1.0 μL/mL in three replicate plates, based on the resul ts of the preliminary cytotoxicity the test article

. Two UDS and parallel cytotoxicity assays were conducted with nine doses of the test material (0.0001 to 1.0 μL /mL)

. Because of unspecified technical difficulties, the first assay was terminated.

In the repeat assay, the two highest test doses (0.3 and 1.0 μL/mL) induced marked cytotoxicity (85%) and were, therefore, not scored for UDS activity

. The five levels selected for the evaluation of UDS induction were 0.001, 0.003, 0.01, 0.03, and 0.1 μL/mL

. The highest test material dose scored for UDS (0.10 μL/mL) induced a marked increase in the net nuclear grain count compared to the solvent control

. Similarly, the percentage of cells with ≥5 net nuclearDocket Number EPA

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P age 47 of 74 grains (NGC) was markedly higher at this level compared to the solvent control value

. However, the evidence for a genotoxic effect was confined to the 0.10 μL/mL dose group

. The mean net nuclear grain counts and percentage of cells with ≥5 net nuclear grains for hepatocytes exposed to 0.001, 0.003, 0.01, or 0. 03 μL/mL of the test material, although higher than the solvent control, did not indicate a positive or a dose

- related response

. The study author concluded that hexahydro

- l,3,5

- tris(2

- hydroxyethyl)

- s

- triazine induced an equivocal positive response in this test system.
This study is classified as Una cceptable

. In the absence of a dose

- related effect, confirmation of a “significant” increase at a single dose is necessary to conclude a positive response in the UDS
assay

. However, the positive response was not dose related, was limited to the 0.l0 μL/mL treatment group, and the results were not confirmed

. In addition, the study author failed to provide information on test material purity, the batch number of the assayed sample, and analytical data to verify act ual concentrations used in the assay

. Therefore, it was concluded that the study is unacceptable, but the test material is classified as presumptively genotoxic in this test system.

(2) In an unscheduled DNA synthesis (UDS) study (MRID 43020001), following preliminary cytotoxicity testing (by lactate dehydrogenase, LDH, release determinations), triplicate coverslip cultures (per does) of rat hepatocytes were exposed to hexahydro

- l, 3, 5

- tris(2

- hydroxyethyl)

- s

triazine for 18

- 20 hours to a series of nine con centrations of test article (ranging from 0.001 to 0.30 μg/mL), together with a constant dose of tritiated thymidine (HTdr, 10 μCi/mL)

. Other cultures were treated with 7, 12

- dimethylebenz(a)anthracene (DMBA, 3 and 10 μg/mL), to serve as positive controls

. For the UDS assay, examination of the fixed and stained cells which had been treated at doses of 0.15, 0.2, 0.25, and 0.3 μg/mL revealed that these could not be evaluated for UDS “because of excessive cytotoxicity”, resulting in relative cell survivals l ess than 35%

. Although none of the remaining (lower) test article doses (0.01, 0.03, 0.05, 0.07, and 0.1 μg/mL) caused a significant increase in mean NNC, according to the authors’ criteria established for declaring a positive result (i.e., an increases of at least five counts over solvent control), there was a dose

- related increase trend in both mean NNC, as well as percentage of cells showing DNA repair

. On the other hand, DMBA

- treated cultures showed a definitively significant mean NNC (>5 counts) in 99% of cells in repair

. Hence, the authors concluded that the test article was not considered positive according to their criterion for a genotoxic response.

This study is classified as Unacceptable , though “ equivocally (presumptively) positive ”, because of slight, but dose

- related, increases in both mean net nuclear silver grain counts (NNG), as well as percentage of cells in repair at the usable three highest dosage levels 0.05, 0.07 and 0.10 μg/mL, but which did not reach the criterion for significance pro posed by the investigators, namely, increase of 5+ counts over control

. Higher doses were stated to be “too toxic to be evaluated for UDS” (as suggested by LDI4 release at 30

- 60% lethality), necessitating in the reviewer’s opinion a repeat assay to resolve this equivocation, together with (l) a finer sequence of dosages to recover viable cultures at higher concentrations, (2) testing hepatocytes from a female rat, and (3) providing the purity (% a.i.) of the test article.Docket Number EPA

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P age 48 of 74 In reviewing the data base for this substance, revealed a previous UDS study (MRID 41262301 ), conducted by the same laboratory

. This study was also classified Unacceptable , requiring confirmation of a positive response (increased NNG >5, and increased % cells in repair) at the HDT of 0.10 μ g /ml.

Thus, these two comparable assays, when considered together, constitute the recommended confirmation of singularly positive results, and may be re

- classified ACCEPTABLLE, in ratifying that HHT induces UDS (repair) above background at mildly toxic do ses (<20% lethality).

(3) In an in vivo unscheduled DNA synthesis assay in rat hepatocytes (MRID 47284304), based on the mortality observed in a preliminary acute toxicity test, male and female Wistar rats were orally (gavage) exposed to Protectol HT ( 79% aqueous solution, batch # 1157 ), in deionized water, at concentrations of 0, 375 , or 750 mg/kg (male) and at 0, 250, or 500 mg/kg (female) for a period of either 2 or 16 hours

. Rats were also exposed to two positive mutagens (DMH and 2

- AAF) at concentratio ns of 40 mg/kg and 100 mg/kg, respectively

. Oral administration of Protectol HT did not induce an increase in unscheduled DNA synthesis in male and female rat hepatocytes at any concentration tested

. No increase in the nuclear grains or net grains was obs erved in hepatocyte cultures exposed for a period of 2 or 16 hours

. The positive controls elicited the appropriate responses

. There was no evidence of induction of unscheduled DNA synthesis in rat primary hepatocytes over the background.

This study is clas sified as Acceptable/Guideline and satisf ies the guideline requirement for Test Guideline OPPTS 870.5550; OECD 486 for other genotoxic mutagenicity data.

Neurotoxicity

870.6200 Neurotoxicity Screening Battery

Adequacy of Database for Neur o to xicity: The database for neur otoxicity is considered incomplete . There are no neur o to xicity studies in the Agency’s hazard database. A neurotoxicity screening was conducted in a 90

- day oral (drinking water) toxicity study (MRID 47300901)

, which was summar ized belo w for the neu rotoxicity screening portion of the study.

B ased on the currently registered indirect food (countertop disinfection and dish detergent) and metal

- working fluid (MWF) uses for HHT , an acute neurotoxicity study in rats is anticipated to be requi red for registration review risk assessment

.
In a 90

- day oral toxicity study (MRID 47300901 )

, h exahydro

- 1,3,5

- t ris

- (2

- h ydroxyethyl)

- S

t riazine (79% a.i. , Batch # 1157) was administered to 10 Wistar rats/sex/dose in water at dose levels of 0, 200

, 1000 , or 5000 ppm (equivalent to 0, 14.1

, 64.1

, or 285.2 mg/kg bw/day for males and 0, 21.2, 91.0, or 338.6 mg/kg bw/day for females ).

The following neurotoxicity screening assessment was carried out:

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P age 49 of 74 A functional observation al battery was performed in all animals. The FOB started with passive observations without disturbing the animals, followed by removal from the home cage, open field observations in a standard arena and sensorimotor tests as well as reflex tests. A detaile d description of the methods used , the ranking and documentation system were provided in the study report

. Home cage observations:

The test animals were observed in their closed home cages . A ny disturbing activities (touching the cage or rack, noise)

, during these examinations

, were avoided to eliminate any influence to the behavior of the animals. Attention was paid to:

1) posture 2) tremor 3) convulsions 4) abnormal movements 5) impairment of gait 6) other findings

Open field observations:

The test animals were transferred to a standard arena (50 x 50 cm with sides of 25 cm high) and observed for at least 2 minutes. Following parameters were examined:

1) behavior when removed from cage 2) fur 3) skin 4) salivation 5) nose discharge 6) lacrimati on 7) eyes/pupil size 8) posture 9) palpebral closure · 10 ) respiration 11) tremors 12) convulsions 13) abnormal movements· 14) impairment of gait 15) activity/arousal level 16) feces: (number of fecal pellets/appearance/consistency) within two minutes 17) urine· within two minutes 18) number of rearings within two minutes

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P age 50 of 74 The test animals were removed from the open field and subjected to the following sensorimotor or reflex tests:

1) approach response 2) touch response 3) vision ("visual placing response") 4) pupillary reflex 5) pinna reflex 6 ) audition ("startle response") 7) coordination of movements ("righting response") 8) behavior during "handling" 9) vocalization 10 ) pain perception ("tail pinch") 11) grip strength o f forelimbs 12) grip strength of hindlimbs 13) landing foot

- splay test 14) other findings

Motor activity assessment

Motor activity was measured on the same day as FOB was performed. The measurement was performed in the dark using the Multi

- Varimex

- System ( Columbus Instruments Int. Corp., Ohio, USA) with 4 infrared beams per cage. During the measurement the test animals were placed, in a randomized order , and kept in Polycarbonate cages with absorbent material. The measurements started at about 2.00 p.m. a nd the number of beam interrupts were counted over 12 intervals, each lasting 5 minutes. Measurement did not comme nce at the same instant for all cages; the period of assessment for each animal started when the first beam was interrupted by pushing the cag e into the rack (staggered start). Measurements ended exactly 60 minutes thereafter. N o food and no water were provided to the test animals during the
measurement

. The neurological observations conducted in this 90

- day drinking water study revealed no signs of a dose

- dependent relationship. However, no statistical tests were performed on the neurological data.

The LOAEL is 5,000 ppm ( 285.2 mg/kg body weight/day in males; 338.6 mg/kg body weight/day in females) , based on decreased body weigh t and body weight gain, food and water consumption, and changes in clinical urinary and blood chemistry

. The NOAEL is 1,000 ppm (64.1 mg/kg body weight/day in males; 91.0 mg/kg body weight/day in females)

.
This 90

- day oral toxicity study in the rat is A cce ptable /G uideline and satisfies the guideline

r equirement for a 90

- day oral toxicity study (OPPTS 870.3100; OECD 408) in rats

. The neurotoxicity screening conducted in this 90

- day drinking water study was deemed acceptable and provided qualitative information of potential neurotoxicity of HHT

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P age 51 of 74 

## Immunotoxicity

870.7800 Immunotoxicity Study

Adequacy of Database for Immunotoxicity: The database for i mmunotoxicity is considered incomplete

. There are no i mmunotoxicity studies in the Agency ’s hazard databa se

. An i mmunotoxicity study is anticipated to be required for registration review risk assessment

, based on the currently registered uses for HHT. 

## Other Toxicological Effects

There are two in vitro studies submitted for HHT, one is a dermal penetration st udy using human skin and the other is a metabolism study using human blood and cryopreserved hepatocytes

. Although the in vitro dermal penetration study using human skin is an acceptable study, only one in vitro study does not adequately address dermal absorption of HHT

. However, there are subchronic dermal studies; therefore, an in vivo dermal penetration study is not expected to be required

. Brief summary of these studies are provided below.

(1) In a n in vitro dermal penetration study ( MRID 47353801; also MRID 47281206 ) , [ 14 C]

- formaldehyde

- Hexahydro

- 1,3,5 tris(2

- hydroxyethyl)

- s

- triazine (HHT) ( 99.0% ; batch # 209740/05 [ non

- radiolabelled ] ; 84.30%, 210405/011 [radiolabelled] ) was administered at a dose of 9.91 mg/cm 2 to 8 human skin preparations (split

- th ickness membranes) (Experiment 1) or 7.95 mg/cm 2 to 4 human skin preparations (Experiment 2) that were mounted in diffusion cells occluded with a carbon trap and carbon filter

. Receptor fluid was collected at hourly intervals from 0

- 8 hours post

- application (Experiments 1 and 2), and every 2 hours from 8

- 24 hours post

application (Experiment 1 only)

. The skin was washed and wiped 8 hours after application (Experiments 1 and 2)

. The skin was tape stripped 8 hours after application (Experiment 2) or 24 hours following application (Experiment 1)

. Carbon filters and traps were removed and extracted at 8 hours (Experiments 1 and 2) and 24 hours (Experiment 1).

Recovery of the applied dose (mass balance) in Experiment 1 was unacceptable, ranging fro m 63.75% to 78.57%

. T hi s low mass balance was possibly due to volatile ( [ 14 C]

- formaldehyde) components in the carbon trap

. A second experiment (Experiment 2) was performed to investigate the possible volatility of the test article

. Recovery of the applied dose in Experiment 2 was acceptable in 3 out of the 4 samples

. In both experiments, the highest levels of radioactivity were recovered from the tissue swabs after 8 hours of exposure : 38 and 40% of the applied dose in Experiment 1 and Experiment 2, respec tively

. The next highest levels of radioactivity were present in the skin wash and carbon filter and trap

. These three metrics combined with the radioactivity of the pipette tip, cell wash, unexposed skin, and tape strips yielded the total unabsorbed dose: 68 and 90% of the applied dose in Experiment 1 and 2 respectively

. The total absorbed dose was 2.87% of the applied dose in Experiment 1 and 2.54% percent of the applied dose in Experiment 2

. Based on radio Mass

- Spectrometry, HHT completely degraded in re ceptor fluid and skin samples.

This in vitro study using human skin is A cc eptable/Non

- guideline

. This study is considered

non

- guideline because it is an in vitro human skin penetration and distribution study. This studyDocket Number EPA

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P age 52 of 74 does not satisfy the guideline requirements of OPPTS 870.7600 in vivo dermal penetration study conducted on rats.

(2) In a n in vitro metabolism study ( MRID 47353802; also MRI D 47281205 ) , hexahydro

- 1,3,5

- tris(2

- hydroxyethyl)

- S

- triazine (radiolabeled test i tem batch #: 210405/012, 81.72% radiochemical purity; non

- radiolabeled test item batch #: 209740/05, 99% purity) was administered to male human blood and cryopreserved hepatocytes at dose levels of 0 or 6.6 μM.

Radioactive recoveries were determined

. The c hromatograms of non

- radiolabeled hexahydro

1,3,5

- tris(2

- hydroxyethyl)

- S

- triazine confirmed retention time of the parent compound as 10 minutes and radiolabelled hexahydro

- 1,3,5

- tris(2

- hydroxyethyl)

- S

- triazine showed an impurity eluting at 3.5 minutes

. Sinc e this impurity was greater than the impurity observed in the parent compound, it suggests instability in this study (incubation matrix, stopping solution, and/or mobile phase)

. Chromatograms for media, hepatocytes, and blood showed no parent compound

rema ining, which suggests instability in this study; therefore, it is not possible to establish whether any metabolism of hexahydro

- 1,3,5

- tris(2

- hydroxyethyl)

- S

- triazine occurred

.
This in vitro metabolism study in human cells is classified Unacceptable / Nong ui deline and does not satisfy the guideline requirement [OPPTS 870.7485, OECD 417] because the test system was not appropriate, the test material was unstable, and the study greatly deviated from the guidelines

.
Table A 2

– Toxicity Profile for HHT

Guideline No./ Study Type MRID No./

Study Classification Dosing and Animal Information Results Subchronic Toxicity 870.3100 90

- day Oral (Rat) MRID 41483001

Minimum Purity 78.5%

Rats (10/sex/group)
0, 10, 50, 100, or 250 mg/kg/day (gavage) NOAEL = 50 mg/kg/day LOAEL = 100 mg/kg/day , based on lymphocytic infiltration in females and erosion of gastric mucosa and prominence of limiting ridge of the stomach in males.
870.3100 90

- day Oral (Rat) MRID 47300901

Acceptable/Guideline Purity 79%

Rats (10/sex/group)
0, 200, 1000, or 5000 ppm (equivalent to ♂ : 0, 14.1, 64.1, or 285.2 mg/kg bw/day; ♀ : 0, 21.2, 91.0, or 338.6 mg/kg bw/day) in drinking water NOAEL = 1000 ppm (♂ : 64.1 mg/kg/day; ♀ : 91.0 mg/kg/day) LOAEL = 5000 ppm (♂ : 285.2 mg/kg/day; ♀ : 338.6 mg/kg/day) , based on lymphocytic infiltration in females and erosion of gastric mucosa and prominence of limiting ridge of the stomach in males.Docket Number EPA

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P age 53 of 74 Guideline No./ Study Type MRID No./

Study Classification Dosing and Animal Information Results

870.3200 21

- day Dermal (Rat) MRID 00155989/260195

Minimum Purity 78.5%

Rats (5/sex/group)

0, 100, 500, or 1000 mg/kg/day NOAEL dermal < 100 mg/kg/day LOAEL dermal = 100 mg/kg/day (lowest dose tested [LDT]), based on multifocal scab formation at treatment sites and histological dose

related findings ( epidermal ulceration, acanthosis and dermal inflammat ory infiltration with occasional necrosis and hemorrhage ).

NOAEL systemic ≥ 1000 mg/kg/day (highest dose tested [HDT]) LOAEL systemic not established

. 870.3250 90

- day Dermal Subchronic (Rat) MRID 41858301

Minimum Purity 96.6%

Rats (13sex/group)

0, 10, 30, or 100 mg /kg/day NOAEL dermal < 10 mg/kg/day (lowest dose tested [LDT]) LOAEL dermal = 10 mg/kg/day , based on the gross observation of dermal irritation and histological dermal lesions (hyperplasia, hyperkeratosis, inflammation, sebaceous gland hyperplas ia and eschar formation).

NOAEL systemic = 30 mg/kg/day LOAEL systemic = 100 mg/kg/day

, based on severe dermal irritation in both sexes and decreased body weight reported in high dose males.
870.3250 90

- day Dermal Subchronic (Rat) MRID 41483002

Minimum Purity 78.5%

Rats (10/sex/dose)

0, 5, 50, 250 mg/kg NOAEL dermal = 5 mg/kg/day LOAEL dermal = 50 mg/kg/day , based on the presence of skin lesions

NOAEL systemic > 250 mg/kg/day (highest dose tested [HDT]) LOAEL systemic not established

. 870.3465 90

- day In halation (Rat) A subchronic inhalation toxicity study is not available for HHT.

Developmental Toxicity 870.3700a Developmental Toxicity (Rat) MRID 41161801

Minimum Purity 78.5%

Rats (24/dose)

0, 250, 500, or 750 mg/kg/day, GD 6

- 15 NOAEL maternal = 500 mg/kg/day LOAEL maternal = 750 mg/kg/day

, based on decreased body weight gain, ulcerations, and scarring of the stomach mucosa.

NOAEL developmental ≥ 750 mg/kg/day (highest dose tested [HDT]). LOAEL developmental not established

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P age 54 of 74 Guideline No./ Study Type MRID No./

Study Classification Dosing and Animal Information Results

870.3700a Developmen tal Toxicity (Rat) MRID 41865701

Supplementary Purity: 96.6%

25 rats/group

0, 10, 75 and 150 mg/kg/day, GD 6

- 15 NOAEL maternal = 75 mg/kg/day LOAEL maternal = 150 mg/kg/day

, based on decreased body weight gain

LOAEL developmental ≤ 10 mg/kg/day (lowest dose tested [LDT]), based on the observed significant positive trend in the incidence of bilateral convoluted ureters and dilated renal pelvises. NOAEL developmental not established

. Reproduction Toxicity 870.3800 Reproduction A reproduction toxicity study is not available for HHT.

Chronic Toxicity 870.4100 Chronic Toxicity A chronic toxicity study is not available for HHT.

Carcinogenicity 870.4200 Carcinogenicity A carcinogenicity study is not available for HHT.

Mutagenicity 870.5100 Bacterial reverse mutation test MRID 41231702

Acceptable/Guideline Purity 78.5%

TA1535, TA1537, TA1538, TA98, or TA100

up to 200 μg/plate with and without S9 metabolic activation Negative

870.5100 Bacterial reverse mutation test MRID 00155991/260195

Unacceptable Purity 78.5%

Strains TA1535, TA1537, TA1538, TA98, or TA100

0, 0.16, 0.8, 4, 20, or 100 μg/plate; ±S9 Positive

Positive in TA96 with activation and in TA1538 with and without activation.

870.5100 Bacterial reverse mutation test MRID 47284305

Unacceptable/Non

guideline (upgradable) Purity not provided

strains TA98, TA100, TA1535, TA1537, of S. typhimurium : 0, 100, 500, 2500, 5000, 7500
 g/plate; ±S9

strains TA98, TA100, TA102; TA1535, TA1537, of S. typhimurium and E.coli WP2 urvA: 0, 25, 50, 100, 200, and 400  g/plate; ±S9 NegativeDocket Number EPA

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P age 55 of 74 Guideline No./ Study Type MRID No./

Study Classification Dosing and Animal Information Results

870.5300 Mammalian cell gene mutation test MRID 47284303

Acceptable/Guideline Purity 79% aqueous solution

Chinese Hamster V79 cells (CHC)

1.6, 3.1, 6.3, 12.5, 25, or 30 μg/mL,

- S9; 1.6, 3.1, 6.3, 12.5, 20, or 25 μg/mL, +S9; for a period of 4 hours Negative

870.5375 In vitro mammalian chromosome aberration test MRID 47284302

Acceptable/Guideline Purity 79%

Chinese Hamster V79 Cells (CHC)

0 , 2.5, 5, 10, 20, 30, or 40 μg/mL; ±S9 for 4 hours Positive

870.5385 In vivo mammalian bone marrow chromosome aberration test (mouse) MRID 41321501

Unacceptable Purity 78.5%

5/sex/group
0, 200, 400, or 800 mg/kg (80% of LD50) Negative

870.5385 In vivo mammalian bone marrow chromosome aberration test (mouse) MRID 41231701

Acceptable/Guideline Purity 78.5%

5/sex/dose
0, 200, 400, and 800 mg/kg; gavage; marrow cells were harvested at 24, 48, or 72 hours post

- treatment. Negative

870.5385 In vivo mammalian bone marrow chromosome aberration test (mouse) MRID 47284301

Acceptable/Guideline Purity 79%

6 male mice/dose
0, 37.5, 75, or 150 mg/kg; intraperitoneally; bone marrow cells were harvested at 24 hours post

- treatment for all dose levels, and 48 hours post

- treatment for the negative control and 150 mg/kg treatment dose

. Negative

870.5385 In vivo mammalian bone marrow chromosome aberration test (rat) MRID 00155992

Unacceptable Rat micronucleus cells

Purity 78.5%
Intragastric intubation, dermal application, or subcutaneous injection ; 2 equal doses, separated by 24 hours, of 15, 60, 240, or 960 mg/kg NegativeDocket Number EPA

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P age 56 of 74 Guideline No./ Study Type MRID No./

Study Classification Dosing and Animal Information Results

870.5550 Unscheduled DNA synthesis MRID 41262301

MRID 43020001

Una cceptable Purity 78.5%

Rat hepatocyte cells

0, 0.001, 0.003, 0.01, 0.03, and 0.10 μ L/mL

0, 0.001, 0.03, 0.01, 0.03, or 0.1
μ g/mL Positive

These 2 studies, when considered together, constitute the recommended confirmation of a singularly positive results, based on HHT inducing UDS (repair) above background at mildly toxic doses (<20% lethality).

870.5550 ( I n vivo ) Unscheduled DNA synthesis MRID 47284304

Acceptable/Guideline Purity 79% aqueous solution

Rat hepatocyte cells ♂: 0, 375, or 750 mg/kg; ♀: 0, 250, or 500 mg/kg; gavage ; either 2 or 16 hours

. Negative

Metabolism 870.7485 (I n vitro ) M etabolism and Pharmacokinetics MRID 47353802 MRID 47281205

Unacceptable Purity 81.72% (radiolabeled);

99% (non

- radiolabeled)

H uman blood and cryopreserved hepatocytes

0

, 6.6 μM T he test system was not appropriate, the test material was unstable, and the study greatly deviated from the guidelines

.

Dermal Penetration 870.7600 ( In vitro ) Dermal penetration MRID 47353801 MRID 47281206

Acceptable/ Non

g uideline Purity 99.0% ( non

- radiolabelled );

84.30% (radiolabelled)

Human skin

9.91 mg/cm 2 to 8 human skin preparations (split

- thickness membranes) (Experiment 1) or 7.95 mg/cm 2 to 4 human skin preparations (Experiment 2) The total absorbed dose was 2.87% of the applied dose in Experiment 1 and 2.54% percent of the applied dose in Experiment 2

. Based on radio Mass

- Spectrometry, HHT completely degraded in receptor fluid and skin samples

.
However, an in vitro study only does not adequately address dermal absorption of HHT.Docket Number EPA

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P age 57 of 74 Toxicology References

MRID

00155959 Consultox Laboratories (1973) Glokill 77: Acute Oral Toxicity. Unpublished

study. 4 p.

00155984 Hughes, T. (1985) Acute Dermal Toxicity Screen in Rabbits of Bioban GK:

Project No. 85

- 0866

- 21. Unpublished study p repared by Hill Top Research, Inc. 10 p.

00155985 Collier, T.; Axelrad, J. (1984) OECD Eye Irritation Test: Determination of the

Degree of Ocular Irritation Caused by Glokill 77 in the Rabbit: Experiment No. 371/8408. Unpublished study pre

- pared by SafePh arm Laboratories Ltd. 11 p.

00155986 Guest, R.; Collier, T. (1985) Primary Dermal Irritation Test: Determination of the

Degree of Primary Cutaneous Irritation Caused by Glokill 77 in the Rabbit: Experiment No. 317/8505. Unpublished study prepared by SafePh arm Laboratories Ltd. 11 p.

00155987 Thomas, M. (1974) Glokill 77: Draize Skin Test, Guinea Pig Sensitization Test:

CL 74: 74: 1024. Unpublished study prepared by Consultox Laboratories Ltd. 8 p.

00155989 Thomas, M.; Brooks, P. (1984) Glokill 77: 21 Day De rmal Toxicity Study in the

Rat: Experiment No. 508/8411. Unpublished study prepared by SafePharm Laboratories Ltd. 89 p.

00155991 Johnson, I. (1984) Ames Test (O.E.C.D.) Product Glokill 77: Project No.

1047/8/J. Unpublished stud y prepared by SafePharm Labora tories Ltd. 25 p.

00155992 Urwin, C.; Richardson, J.; Palmer, A. (1976) An evaluation of the mutagenicity of

the cutting oil preservative Grotan BK. Mutation Research 40:43

- 46.

41161801 Ridgway, P. (1989) Triazine Rat Teratology Study: Reference No. LEF/8/89.

Unpublished study prepared by Toxicol Laboratories Ltd. 117 p.

41231701 Asquith, J.; Phil, M. (1989) Triazine Joint Venture: Mouse Micro

- nucleus Test:

Triazine: Toxicol Study No. M/MMN/10659. Unpublished study prepared by Toxicol Laboratories Lt d. 40 p.

41231702 Asquith, J.; Phil, M. (1989) Triazine Joint Venture: Bacterial Reverse Mutation

Assay: Triazine: Toxicol Study No. M/AMES/10658. Unpublished study prepared by Toxicol Laboratories Ltd. 35 p.

41262301 Curren, R. (1988) Unscheduled DNA Synt hesis in Rat Primary Hepatocytes:

Hexahydro

- 1,3,5

- tris (2

- hydroxyethyl)

- s

- triamine: Study No. T8102.380. Unpublished study prepared by Microbiological Associates, Inc. 27 p.Docket Number EPA

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P age 58 of 74 41321501 Putman, DL and Melhorn, JM (1989) Micronucleus Cytogenetic Assay in Mice.

Final Report. Lab Study No. T8796.122010. Unpublished study prepared by Microbiological Associates, Inc. (Bethesda, MD). November 28, 1989. 28 p.

41483001 Hill, R.; Newman, A. (1990) Triazine: 13 Week Oral (Gavage) Toxicity Study in

the Rat: Lab Project N umber: LEF/3/90: LEF/3/89. Unpublished study prepared by Toxicol Laboratories, Ltd. 253 p.

41483002 Hill, R.; Newman, A. (1990) Triazine: 13 Week Dermal Toxicity Study in the

Rat: Lab Project Number: LEF/4/89. Unpublished study prepared by Toxicol Laborato ries, Ltd. 415 p.

41675206 Rush, R. (1989) Acute Oral Toxicity Study in Rats with Busan 1060: Lab Project

Number: 3138.47. Unpublished study prepared by Springborn Laboratories, Inc. 59 p.

41858301 Trimmer, GW (1991) 90

- Day Subchronic D ermal Toxicity Study in R ats

(VANCIDE TH) . Toxicology Laboratory, Exxon Biomedical Sciences, Inc. (East Millstone, NJ). Exxon ID: MRO

- 89

- 408. Submitted by R. T. Vanderbilt Company, Inc. (Norwalk, CT)

. April 19, 1991. MRID 41858301. Unpublished. 429 pages.

41865701 Beyer, BK ( 1991) Developmental Toxicity Study in Rats with VANCIDE TH.

Toxicology Laboratory, Exxon Biomedical Sciences, Inc. (East Millstone, NJ). Exxon ID: MRO

- 89

- 408. Submitted by R. T. Vanderbilt Company, Inc. (Norwalk, CT). April 26, 1991. MRID 41865701. Unpubli shed. 274 pages.

43020001 San, R.; Raabe, H. (1993) Unscheduled DNA Synthesis in Rat Primary

Hepatocytes: Final Report: Lab Project Number: TC836.380: SPGT380. Unpublished study prepared by Microbiological Associates, Inc. 25 p.

47281205 Findlay, K.; Toner , F. (2007) (Carbon 14)

- HHT: Tissue and Species Variation in

the in

- vitro Metabolism of (Carbon 14)

- HHT. Project Number: 778242. Unpublished study prepared by Charles River Laboratories. 44 p.

47281206 Roper, C.; Stupart, L. (2007) The in

- vitro Percutaneou s Absorption of

Radiolabelled HHT Through Human Skin. Project Number: 776706. Unpublished study prepared by Charles River Laboratories. 70 p.

47284301 Honarvar, N. (2002) Micronucleus Assay in Bone Marrow Cells of the Mouse

with Myacide HT T Application: O nce, Intraperitoneally. Project Number: 711403, 26M0229/009062. Unpublished study prepared by RCC Umweltchemie Gmbh & Co. Kg. 27 p.

47284302 Schulz, M. (2002) In Vitro Chromosome Aberration Test in Chinese Hamster

V79 Cells with Myacide HT T. Project Numbe r: 32M0229/009063, 711401. Unpublished study prepared by RCC Umweltchemie Gmbh & Co. Kg. 35 p.Docket Number EPA

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P age 59 of 74 47284303 Wollny, H. (2002) Gene Mutation Assay in Chinese Hamster V79 Cells in vitro

(V79/HPRT) with Myacide HT T. Project Number: 711402, 50M0229/009064. Unpubl ished study prepared by RCC Umweltchemie Gmbh & Co. Kg. 39 p.

47284304 Honarvar, N. (2002) In vivo Unscheduled DNA Synthesis in Rat Hepatocytes

with Myacide HT T. Project Number: 711404, 80M0229/009065. Unpublished study prepared by RCC Umweltchemie Gmbh & Co. Kg. 34 p.

47284305 Engelhardt, G. (1997) A Study of Myacide HT T Ames Test: (Ames

Salmonella/Mammalian

- Microsome Mutagenicity Test and E.coli/ Mammalian

Microsome Reverse Mutation Assay). Project Number: 40M0271/954200, D/67056. Unpublished study prep ared by BASF Ag Research Station. 43 p.

47300901 Kaspers, U. ( 2002 ) Protectol HT

– Subchronic oral toxicity study in Wistar rats

Administation in drinking water for 3 months. Experimental Toxicology and Ecology, BASF Aktiengellschaft ( Rhein, Germany ). Proj ect No. 52S0229/00128

, March 19, 2002. MRID 47300901

. Unpublished.

47353801 Roper, C.; Stupart, L. (2007) The in vitro Percutaneous Absorption of

Radiolabelled HHT Through Human Skin. Project Number: 776706, 26844. Unpublished study prepared by Charles Riv er Laboratories. 70 p.Docket Number EPA

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P age 60 of 74 

# Appendix B Environmental Fate

Environmental Fate and Transport Properties of Hex a h y dro

- 1, 3, 5,

- tris(hydroxyethyl)

- s - triazine (HHT)

Water and Sediment

Hydrolysis

EPA has reviewed four hydrolysis studies. These studies which are summarized below give conflicting results regarding hydrolysis at high pH. EPA anticipates the need to require a new hydrolysis study to address the rate of degradation and formation of any pertinent degradates

.
1. MRID#: 47353803 : This stud y was conducted as the pH changed from 1 to 11.35 to estimate the stability of HHT at different pHs. The degradation of HHT was followed by changes observed in the HPLC chromatograms. It was o bserved that: 1) At pH 1, HHT deg rad ed instantly (t= 0)

. 2) HHT was stable at pH 11. 35 but less so at pH 7.2

. 3) At pH 4, HHT degraded fast but the degradation time was measureable. The temperature of the study was kept at about 37  ̊ C

. This study showed that HHT is not stable in acidic cond itions, but is stable under basic conditions.

2. MRID# 43181001 : In this study, hydrolytic half

- life measured were: 3.2 days at pH 5, 50 days at pH 7, and 302 days at pH 9. The temperature of the study was 25  ̊ C. Hydrolytic degradate characterized was formald ehyde. Maximum level (percent) of formaldehyde was 100, 31 and 20 at pHs 5, 7, and 9.

3. In the third study (Bakke et al study, 2001 ; open literature study ): it was shown that HHT degraded fast at

–

pH 9.5 and 10.9 and 8.0 (T = 22 C).The half

- life were: 3.4 hours, 16 minutes and 32 seconds respectively. When measured at 60 C, the half

- life were: 3.1 minutes and 6.3 seconds for pH 10.9 and 9.5 respectively. Thus both studies indicate short hydrolytic half

- lives at neutral to basic pHs.

4. MRID# 48741001 : This hy drolysis study in aqueous metalworking f luid was conducted to determine the rate of release of formaldehyde when HHT was used in an aqueous MWF mixture containing 4, 5, and 6% HHT. The study was conducted at 25, and 35  ̊ C. About 30% loss of released formald ehyde was noted. No rationale was provided for 30% loss of formaldehyde. Incomplete study

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P age 61 of 74 Ready Biodegradability:

A ready b iodegradability study by Voets et. al, 1975 on HHT showed that a when 70 mg/L of HHT was tested with 1/10th of its intended use concentration, under aerobic and anaerobic conditions. 100% of HHT biodegraded; f ormaldehyde was the only degradation product measured

. In order to characterize the potential for biodegradation, th e A gency anticipates needing additional information on the formation and decline of degradates.

Adsorption

MRID#: 47281203: A study on the Estimation of Adsorption Coefficient (K oc ) for Mergal KM 200 (a product containing 76% HHT) was conducted on soil and Sewage Sludge using High Performance Liquid /chromatography (HPLC). The study was designed based on th e OECD Test Guideline No. 121.

The results of the study indicated Log K oc could not be determined for Mergal KM 200 ( which contains 76% HHT) as the a ctive showed a peak with the retention that was smaller than the retention time of urea which was used as a reference substance. Study authors made a tentative interpretation: Test substance is highly water soluble, and with very small retention time on th e HPLC column, it could indicate that test substance is very mobile on soil and sewage sludge systems.

Fate and Transport in WWTP

Activated Sludge Sorptio n Isotherm: No data was submitted. Data gap.

Activated Sludge Biodegradation : No Data was submitted. Data gap.

Ready Biodegradability: No Data was submitted. Data gap.Docket Number EPA

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P age 62 of 74 Environmental Fate References

MRID 43181001. Cohen, S. (1994) Hexahydro

- 1,3,5

- tris (2

- hydroxyethyl)

- s

- triazine (Triazine): Hydrolysis of Triazine in Buffered Aqueous Solutions: Final Report: Lab Project Number: ME 9300156. Unpublished study prepared by Univ. of Pittsburgh Applied Research Center. 86 p.

MRID 47281203. Schouten, A.; Verhoef, A. (2005) Estimation of the Adsorption Coefficient (Koc) of Mergal KM200 on Soil and Sewag e Sludge Using HPLC. Project Number: 010/46064/20/01. Unpublished study prepared by TNO Analytical Services. 24 p.

MRID 47353803 Cairns, S.; Davidson, J. (2005) Development and Application of an Informal Method for the Determination and Stability of HHT ( Triazine). Project Number: 25987, TC1916, 209159. Unpublished study prepared by Inveresk Research International. 18 p.

MRID 48741001. Schuchardt, S. (2011) Troyshield B2: Hydrolysis in Aqueous Metal Working Fluid (MWF)

- Determination of the Formaldehyde Release. Project Number: 15G10032. Unpublished study prepared by Fraunhofer Institute of Toxikology and Experimental Medicine. 48p.

Voets, JP, Pipyn, P, Van Lanker, P, and Verstraete, W: 1975: Degradation of Microbiocides under Different Environmental Co nditions, J. Applied Bact.: 40:67

- 72

Bakke, JM, Buhaung J

. and Riha, J, 2001. Hydrolysis of, 1, 3, 5

- Tris(2

- hydroxyethyl)hexahydro

s

- triazine and its reaction with H 2 S. Department of Chemistry, Norwegian University of Science and Technology. Ind. Eng. Chem . Res. 40: 6051

- 6054Docket Number EPA

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P age 63 of 74 

# Appendix C Ecotoxicology Profile

Toxicity to Terrestrial Organisms

Birds

The Agency requires one acute

- oral study to establish the toxicity of HHT (technical grade active ingredient, TGAI) to birds for all registered uses. The preferred test s pecies is either the mallard ( Anas platyrhynchos ) or the northern bobwhite ( Colinus virginianus ). One available acute

- oral study indicates that HHT is only slightly toxic to birds (Table C 1). Avian dietary toxicity studies (northern bobwhite and mallard) are conditionally required for antimicrobial pesticides

, depending on the avian acute

- oral toxicity, pertinent environmental fate characteristics , and a potential for exposure

. Although not required for the current uses of HHT
, the three dietary studies pr eviously submitted categorize HHT as being practically nontoxic when ingested (Table C 1). An avian precautionary statement is not required on product labels. The guidelines for avian
acute

- oral toxicity (OPPTS 850.2100) and avian dietary toxicity OPPTS 850 .2200) are satisfied

.
Tabl e C 1

. Acute

- oral and Dietary Toxicity of HHT to Birds

Test Species Test

Type % ai

tested Toxicity

Endpoint Toxicity

Category Study

Status MRID

No.

Northern bobwhite acute

oral 83.8 LD50 = 1520 (mg ai/kg bw) slightly

toxic core 43154301

dietary 83.8 LC50 >5620 (ppm) practically

nontoxic core 43154302

dietary ?

(Milidin X

- 2) LC50 >5000

(ppm) practically

nontoxic supplemental 240113101

Mallard dietary ? (Milidin X

- 2) LC50 >5000

(ppm) practically

nontoxic supplemental 240113101

Nontarget Insects

Toxicity data for beneficial insects (850.3020, 850.3030) are not expected to be required for the use patterns of HHT

. Mammals, Acute and Chronic

Refer to the human toxicology chapter for details on the available mammalian toxicity studies

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### Toxicity to Aquatic Organisms

Freshwater Fish

Freshwater Fish, Acute

The Agency requires o ne acute toxicity study with the TGAI to establish the toxicity of HHT to freshwater fis h. The preferred test species would be either the rainbow trout ( Oncorhynchus mykiss ), a coldwater fish, or the bluegill ( Lepomis macrochirus ), a sunfish. For HHT , acute studies are available for both the rainbow trout and the bluegill and for the channel catfish ( Ictalurus punctatus ) (Table C 2). The acute toxicity values from these studies categorize HHT as being slightly toxic to freshwater fish

. A precautionary label statement is not required. The guideline ( OPPTS 850.1075 ) is satisfied.

Table C 2

. Acute Toxicity of HHT to Freshwater Fish

Test Species % ai

tested 96

- h LC50

(mg ai/L) Toxicity

Category Study Status MRID No.

Rainbow trout 65.9

(Triadine 10) 41.7 slightly toxic supplemental 247712

? (Milidin X

- 2) 71 slightly toxic supplemental 248883

? (Milidin X

- 2) 74 slightly toxic supplemental 240113106

Bluegill ?

(Milidin X

- 2) 39 slightly toxic supplemental 248883

? (Milidin X

- 2) 39.8 slightly toxic supplemental 240113106

65.9 (Triadine 10) 77 slightly toxic supplemental 247712

Channel catfish 65.9

(Triadine 10) 36 slightly toxic supplemental 247712

The Agency requires one study with the TGAI to establish the chronic toxicity of HHT to freshwater fish. The preferred test species is the bluegill sunfish. Data are anticipated to be needed for this guideline ( fish early

- life stage, 850.1400).

Freshwater Invertebrates

The Agency requires one study with the TGAI to establish the acute toxicity (EC50) of HHT to freshwater invertebrates. The preferred test species is Daphnia magna , a water flea . Results ofDocket Number EPA

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P age 65 of 74 two submitted studies a re presented in Table C 3. Based solely on mortality (LC50), the two studies categorize HHT as being moderately toxic to freshwater invertebrates

. However, an EC50 (based on immobility of exposed daphnids) was not determined in either study. The guideline r equirement (OPPTS 850.1010) is satisfied for current uses. However, a study establishing an EC50 (based on mortality and immobility) may be required to support any new uses in which HHT is expected to reach the aquatic environment at exposure levels of con cern for aquatic organisms

.
Table C 3

. Acute Toxicity of HHT to Freshwater Invertebrates

Test Species % ai

tested 48

- h LC50

(mg ai/L) Toxicity

Category Study Status MRID No.

Water flea 65.9

(Triadine 10) 5.4 moderately toxic supplemental 247712

83.8 26.1 slightly toxic supplemental 43175401

The Agency requires one study with the TGAI to establish the chronic toxicity of HHT to freshwater invertebrates. The preferred test species is Daphnia magna , a water flea . Data are anticipated to be needed for this guideline ( aquatic invertebrate life cycle, 850.1300 ).

Estuarine/Marine Fish and Invertebrates

The Agency requires a cute toxicity testing with estuarine and marine organisms using the TGAI when the end

- use product is intended for direct application to the marine/estuarine environment or the active ingredient is toxic to aquatic organisms and is expected to reach this en vironment via other transport pathways. The preferred test species are the sheepshead minnow ( Cyprinodon variegatus ), mysid shrimp ( Mysidopsis bahia ), and Eastern oyster ( Crassostrea virginica )

. Table C 4 presents available data support ing HHT drilling uses in the estuarine/ marine environment. Test results indicate that HHT is practically nontoxic to moderately toxic to estuarine/marine organisms. The guidelines (OPPTS 850.1025, OPPTS 850.1035, OPPTS 850.1075) are satisfied

.
Table C 4

. Acute Toxicity of HHT to Estuarine/Marine Invertebrates

Test Species % ai

tested 96

- h LC50 or EC50

(mg ai/L) Toxicity Category Study Status MRID No.

Sheepshead minnow 83.8 >118 practically nontoxic supplemental 43143102

Eastern oyster 83.8 2.3 (shell deposition) moderately toxic core 43175402

Mysid shrimp 83.8 12 slightly toxic supplemental 43143103

Aquatic PlantsDocket Number EPA

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P age 66 of 74 No data are available. Studies expected to be needed for the TGAI including a toxicity test with a vascular species (850.4400) and a lgal toxicity test s (850.45 00 and 850.4550 ) with four species ( Anabaena flos

- aquae, Navicula pelliculosa, Skeletonema costatum, and Selenatrum capricornutum ).

Sediment T oxicity

Based on the environmental

- fate properties of HHT , sediment toxicity data are not expected to be required.

Ecotoxicology ReferencesDocket Number EPA

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P age 67 of 74 Campbell, S.; Beavers, J. (1994) Triazine: An Acute Oral Toxicity Study with the Northern Bobwhite: Lab Project Number: 210/119. Unpublished study prepared by Wildlife International Ltd. 19 p

. MRID Campbell, S.; Beavers, J. (1994) Triazine: A Dietary LC50 with the Northern Bobwhite: Lab Project Number: 210/118. Unpublished study prepared by Wildlife International Ltd. 18 p

. MRID 43154302

Carson, S. (1968) The Effect of Grotan on Fingerling Fish: Laboratory No. 88 969. (Unpublished study received Jun 13, 1977 under 1839

- 59; prepared by Food and Drug Research Laboratories, Inc., submitted by Onyx Chemical Co., Jersey City, NJ; CDL:240105

- A)

. MRID 123909

Davis, J.; Kernaghan, N. (1994) Acute Toxicity to Rainbow Trout, Oncorhynchus mykiss , under Flow

- Through Test Conditions: Triazine: Lab Project Number: J9306004E. Unpublished study prepared by Toxikon Environmental Sciences. 46 p

. MRID 43143101

Davis, J. (1994) Acute Toxicity to the Water Flea, Daphnia magna , under Flo w

- Through Test Conditions: Triazine: Lab Project Number: J9306004D. Unpublished study prepared by Toxikon Environmental Sciences. 47 p

. MRID 43175401

Davis, J. (1994) Acute Toxicity to the Mysid, Mysidopsis bahia , under Flow

- Through Test Conditions: Triazine: Lab Project Number: J9306004F. Unpublished study prepared by Toxikon Environmental Sciences. 46 p

. MRID 43143103

Davis, J. (1994) Acute Effect on New Shell Growth of the Eastern Oyster, Crassostrea virginica : Triazine: Lab Project Number: J930600 4H. Unpublished study prepared by Toxikon Environmental Sciences. 49 p

. MRID 43175402

Fletcher, D. (1975) Report to De Mille Chemical Corporation: 8

- Day Dietary LCI50^ Study with Milidin X

- 2 in Mallard Ducklings: IBT No. 651

- 07430. (Unpublished study recei ved Sep 29, 1975 under 21167

- 2; prepared by Industrial Bio

- Test Laboratories, Inc., submitted by Demille Chemical Corp., Jersey City, N.J.; CDL: 227630

- A)

. MRID 47528

Fletcher, D. (1973) Report to Demille Chemical Corporation: 8

- Day Dietary LCI50^ Study wi th Milidin X

- 2 in Bobwhite Quail: IBT No. 651

- 03691. (Unpublished study received May 14, 1964 under 21167

- 2; prepared by Industrial Bio

- Test Laboratories, Inc., submitted by Demille Chemical Corp., Jersey City, N.J.; CDL: 227619

- A)

. MRID 47532

LeBlanc, G.; Sousa, J. (1982) Acute Toxicity of Triadine 10 to Channel Catfish ...: Bionomic s Report #BW

- 82

- 2

- 1120. (Unpub lished study received Jun 11, 1982 under 1258

- 990; prepared by EG & G, Bionomics, submitted by Olin Corp., Stamford, CT; CDL: 247712

- A)

. MRID 1051 89

LeBlanc, G.; Sousa, J. (1982) Acute Toxicity of Triadine 10 to Rainbow Trout ...: Report #BW

82

- 2

- 1117. (Unpublished study received Jun 11, 1982 under 125 8

- 990; prepared by EG & G, Bio nomics, submitted by Olin Corp., Stamford, CT; CDL:247712

- B)

. MRID 10 5190Docket Number EPA

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P age 68 of 74 LeBlanc, G.; Sousa, J. (1982) Acute Toxicity of Triadine 10 to Bluegill ...: Bionomics Report #BW

- 82

- 2

- 1119. (Unpublished study received Jun 11, 1982 under 1258

- 990; prepared by EG & G, Bionomics, submitted by Olin Corp., Stamford, CT; CDL:247712

- C)

. MRID 105191

LeBlanc, G.; Sousa, J. (1982) Acute Toxicity of Milidin X

- 2 to Rainbow Trout ...: Bionomics Report #BW

- 82

- 4

- 1145. (Unpublished study received Nov 8, 1982 under 21167

- 2; prepared by EG & G, Bionomics, submitted by Demille Chemical Corp., Jersey City, NJ; CDL:248883

- A)

. MRID 118414

L eBlanc, G.; Sousa, J. (1982) Acute Toxicity of Milidin X

- 2 to Bluegill ...: Bionomics Report #BW

- 82

- 4

- 1144. (Unpublished study received Nov 8, 1982 under 21167

- 2; prepared by EG & G, Bionomics, submitted by Demille Che mical Corp., Jersey City, NJ; CDL:248883

- B)

. MRID 118415

LeBlanc, G.; Surprenant, D. (1982) Acute Toxicity of Triadine

- 10 to the Water Flea ...: Bionomics Report #BW

- 82

- 2

- 1122. (Un

- published study received Jun 11, 1982 under 1258

- 990; prepared by EG & G, Bionomics, submitted by Olin Corp., Stamford, CT; CDL:247712

- D) MRID 105192

Rausina, G. (1973) Report to Demille Chemical Corporation: Four

- Day Static Fish Toxicity Studies with Milidin X

- 2 in Rainbow Trout and Bluegills: IBT No. 665

- 03509. (Unpublished st udy received May 14, 1974 under 21167

- 2; prepared by Industrial Bio

Test Lab

- oratories, Inc., submitted by Demille Chemical Corp., Jersey City, N.J.; CDL:227625

- A)

. MRID 73588

Ward, S. (1994) Acute Toxicity to the Sheepshead Minnow, Cyprindon variegatus

, under Flow

Through Test Conditions: Triazine: Lab Project Number: J9306004G. Unpublished study prepared by Toxikon Environmental Sciences. 44 p

. MRID 43143102Docket Number EPA

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# Appendix D Screening Level Down

- the

Drain Analysis

A screening

- level Down

- the

- Drain analysis of HHT was not performed since such an analysis is appropriate only for uses of antimicrobial pesticides that would be released solely to domestic wastewater treatment facilities. Uses of HHT include industrial use s that would be expected to have the potential to release HHT and/or its degradates to industrial wastewater treatment facilities and in

- service uses that would be expected to have the potential to release HHT and/or its degradates to domestic wastewater treatment facilities. Since HHT has the potential to be released to both domestic and industrial wastewater treatment facilities, results from a screening

level down

- the

- drain analysis would provide incomplete information on the potential for risks to aquatic organisms from exposure to HHT and/or its degradates.Docket Number EPA

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# Appendix E Product Chemistry

The HHT product chemistry information is summarized in Table E1 (source: MRIDs 47284307, 47284308, and EPI Suite v4.1).

Table E1

– Prod uct Chemistry of HHT

Guideline Numbers HHT Source

830.1800 Enforcement analytical method HPLC

830.6302 Color The color varies from a light transparent brown to an almost clear colorless liquid. The product darkens with age.

830.6303 Physical State Physical State

- The TGAI and end

- product are one in the same and this is a liquid. The TGAI is not a solid.

830.6304 Odor Aldehyde like 830.6313 Stability Stable for one year 830.6314 Oxidation/Reduction Not applicable 830.6315 Flammability Not applicable 830.6316 Explodability Not applicable 830.7000 pH at25 °C Not applicable 830.7050 UV/Visible Absorption Not applicable

830.7200 Melting point (°C) 155 o C

830.7220 Boiling point (°C) Not applicable 830.7300 Density ( g/cm 3 ) 1.2

- 1.6 g/L

830.7370 Dissociation Constant pKa(s) Not applicable

830.7550 Log Kow - 2.3 (pH 5, 24 C)

- 2.0 (pH 7, 24 C); 1.3 (pH 9, 24 C) MRID# 47284308

830.7840 Solubility in water (mg/L) at 25 o C 3 g/L (EPI Suite) 830.7860 Solubility in organic solvents Not applicable

830.7950 Vapor pressure (mm Hg) at 25 °C 11.2 mm Hg at 20 C; 15.15 mm Hg at 25 C; 61.1 mm Hg at 50 C MRID 47284307Docket Number EPA

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P age 71 of 74 Product Chemistry References

EPI Suite v4.1

MRID 47284307. Hutmacher, H. (2002) Vapour Pressure of Myacide HT T: Final Report. Project Number: 01L00575. Unpublished study prepared by BASF Aktiengesellschaft, Labor fuer Oekotoxicologie. 12 p.

MRID 47284308. Dogemuller, A. (2002) Partition Coefficient n

- Octanol/Water [log P(ow)] of Myacide HT T: Final Report. Project Number: 01L00533. Unpublished study prepared by BASF Aktiengesellschaft, Labor fuer Oekotoxicologie. 15 p.Docket Number EPA

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# Appendix F Agency Response to Public Comments

During the public comment period for the PWP for HHT the Agency received two public
comments summarized below.

1. Comment submitted by Adrian Krygsman ( Director, Product Registration, Troy Chemical Corporation )

.

General comments on the PWP :

a. Appendix E: Product Chemistry: i. Color

- our product is not brown to dark brown in color. The color varies from a light transparent brown to an almost clear colorless liquid. The product darkens with age. ii. Physical State

- The TGAI and end

- product are one in the same and this is a liquid. The TGAI is not a solid.

Agency Response: The Agency confirms the information in its files and changes have been added to Appendix E.

b. Occupational Exposure: i. Table 8, footnote 2

- the footnote incorrectly indicates outdoor data are required to address this issue.

Agency Response: The agency accepts the comment and will require only in door data for dermal and inhalation exposure data as this would bracket the worst case
exposure.

Risk Assessment Concerns :

a. New data and models are now available which should be incorporated into a new and updated risk assessment for this compound.

Agency Response: The Agency will conduct its r isk a ssessment using the most recently accepted models and data.

b. New data have been developed which addressed formaldehyde inhalation toxicity.

Agency Response: Thank you for the additional information

. The Agency will consider all available data

, including the Biologically Based Dose Response Model (BBDR) by the Hamner Institute referenced in the comment

, during the registration review risk assessment

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P age 73 of 74 c. There are weaknesses with the methods used for the two studies on occupational exposure with metal working fluids associated with HHT.

Agency Response: The Agency agrees that the risk assessment could be improved if updated exposure studies on metal working fluid were submitted. At the time of the RED t he Agency used a monitoring study submitted by the registrants (MRID 4733544

- 02) and a study from the literature ( Li nnainmaa

, 2003 )

. Th ese studies w ere the best available at the time. The specific limitation s mentioned in the Agency

r eview of these studie s were outlined in revised occupational and residential risk assessment (D 346240). We encourage the registrant to submit new exposure data during registration review to refine the risk estimates made in the RED

. In addition to the information provided in the ORE assessment for the RED , it might be helpful if air sampling were conducted using a filter in front of the sampling tube to prevent the creation of formaldehyde artifacts from the derivatization of MWF aerosols containing unhydrolyzed HHT.

d. The Agenc y and industry need to meet and agree on methods of analysis for data development of HHT and it ’ s degradates.

Agency Response: The Agency is prepared to meet with registrants and stakeholders to receive information that impacts registration review and recommends submitting a protocol prior to data generation

.

2. Comment submitted by William Woods ( Business Manager, Tri

- K Industries Performance Products )

.

a. The commenter q uestioned the 100% conversion of “bound” and “free” formaldehyde in the RED. Tri

- K intends to provide the Agency with simulated exposure studies that can be used to determine the appropriate formaldehyde conversion factor for HHT.

Agency Response: The Agency will consider the simulated exposure studies; in addition, EPA will call in da ta on hydrolysis to be submitted in registration review

. The data should help refine the initial conservative assumptions of 100 percent conversion of HHT to formaldehyde which were made in the 2008 RED.

b. The commenter r equest ed a temporary hold on toxicolo gy data requirements 870.3150; 870.3465; 870.3700; 870.3800; 870.4100 and 870.4200 until the hydrolysis behavior of HHT is fully and adequately characterized.

Agency Response: Required studies identified in a DCI notice must be submitted according to the specified timeframes

, and the timeframes for the toxicology requirements will not be exten d ed at this time

. The Agency notes that t hese studies were identified as data gaps in the RED, and the post

- RED Data

- Call

- In (DCI) was issued on May 12, 2010 and none of these studies have been submitted to the Agency. These studies are required, based on the currently registered uses of HHT, and criticalDocket Number EPA

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for conducting risk assessment for HHT Registration Review

, otherwise the worst case scenario may be used. Alth ough a hydrolysis study provides information on how HHT behaves in the environment, it does not necessarily indicate that HHT will behave similarly in the biological system. Therefore, the Agency is revising the anticipated data needs to include a metaboli sm/pharmacokinetics study to evaluate the absorption, distribution, biotransformation, and excretion of pesticides, to help better understand how HHT behaves in the mammalian system.

c. The commenter r equest ed that the Agency include in the FWP the use of HHT in residential detergent and cleaning products.

Agency Response: These uses were not included in the preliminary work plan because the Agency expected all registrants to comply with the mitigation specifie d in the RED, including the removal of uses and the reduction of application rates. The registrant of the last remaining product with uses that were ineligible for reregistration has indicated its intent not to add the mitigation measures specified in the RED but to now support the uses; therefore, the currently

- registered uses that were found to be ineligible for reregistration as well as the related newly anticipated data needs and related newly anticipated risk assessments have been added to this work pl an. The Agency will also continue to pursue label compliance through regulatory or other action during this registration review.