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Page 2 of 58 HHT Registration Review Team

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

Risk Management T ina Pham Rose Kyprianou Elizabeth Wa tkins Seiichi Murasaki

Office of General Counsel Phil ip RossDocket Number EPA

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Page 3 of 58 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 C hemical 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 Insecticid e, 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 par tition 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 og arithm 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 manufactur ing

- 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 Pr evention OECD Organization for Economic Co

- operation and DevelopmentDocket Number EPA

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Page 4 of 58 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 n egative base

- 10 logarithm of the acid

dissociation constant of a solution ppb p arts p er b illion
ppm p arts p er 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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Page 5 of 58 

### Table of Contents
1 Introduction

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

................................ ................................ ................. 7 1.2 Case Overview

................................ ................................ ................................ ................. 8 1.3 Chemical Identification and Properties

................................ ................................ ............ 9 1.4 Use/Usage Description

................................ ................................ ................................ ... 10 1.4.1 Re gistrations

................................ ................................ ................................ ........... 10 1.4.2 Summary of Registered Uses

................................ ................................ .................. 11 1.4.3 Summary of Risk Mitigation Measures from HHT RED

................................ ....... 11 1.4.4 Usage Information

................................ ................................ ................................ .. 12 1.5 Regulatory History

................................ ................................ ................................ ......... 12 1.5.1 Tolerance Information

................................ ................................ ............................ 12 1.6 Incidents

................................ ................................ ................................ ......................... 12 1.6.1 Human Health

................................ ................................ ................................ ......... 12 1.6.2 Ecological

................................ ................................ ................................ ............... 13 2 Anticipated Data Needs

................................ ................................ ................................ ........ 13 3 Human Health Risk Assessment

................................ ................................ ........................... 17 3.1 Existing Toxi cological Endpoints

................................ ................................ .................. 17 3.2 Dietary Exposure

................................ ................................ ................................ ............ 18 3.2.1 Drinking Water

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

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

................................ ................................ ............. 19 3.3.2 Occupational Post

- application Expos ures

................................ ............................... 19 3.3.3 Residential Handler

................................ ................................ ................................ . 20 3.3.4 Residential Post

- Application Exposures

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

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

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

................................ ................................ ............................ 20 4 Environmental Risk Assessment

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

................................ ................................ .................... 21 4.1.1 W ater Quality

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

................................ .......... 22 4.3 Ecological Effects Assessment

................................ ................................ ....................... 22 4.3.1 Mechanism of Action

................................ ................................ .............................. 23 4.3.2 Measures of Effect (Ecotoxicology Endpoints)

................................ ...................... 23 4.4 Exposure Analysis Plan

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

................................ ............ 24 4.4.2 Screening Level Down

- the

- Drain An alysis

................................ ............................ 25 4.5 Effects Analysis Plan

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

................................ ................................ 25 6 Label Changes

................................ ................................ ................................ ....................... 26 7 Guidance for Commenters

................................ ................................ ................................ .... 26 7.1 Preliminary Work Plan

................................ ................................ ................................ ... 26 7.1.1 Trade Irritants

................................ ................................ ................................ .......... 26 7.1.2 Water Quality

................................ ................................ ................................ .......... 27 7.1.3 Environmental Justice

................................ ................................ ............................. 27Docket Number EPA

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Page 6 of 58 7.1.4 Structure Activity Relationships

................................ ................................ ............. 27 7.1.5 Additional Information

................................ ................................ ........................... 28 8 Next Steps

................................ ................................ ................................ ............................. 28 9 References

................................ ................................ ................................ ............................. 29

Appendix A T oxicology Profile

................................ ................................ .............................. 30 Appendix B Environmental Fate

................................ ................................ ............................. 49 Appendix C Ecotoxicology Profile

................................ ................................ .......................... 51 Appendix D Screening Level Down

- the

- Drain Analysis

................................ ........................ 57 Appendix E Product Chemistry

................................ ................................ ............................... 58

List of Tables Table 1

– Summary of Anticipated Risk Assessm ents and Data Needs for Registration Review: HHT

................................ ................................ ................................ ................................ ................ 8 Table 2

– Anticipated Registration Review Schedule

................................ ................................ .... 9 Table 3

– Chemical Identif ication of HHT

................................ ................................ ..................... 9 Table 4

– Physical

- Chemical and Environmental Fate Properties for HHT

................................ . 10 Table 5

– Summary of HHT Registered Uses

................................ ................................ .............. 11 Table 6

– Risk Mitigation Measures Specified in the HHT RED

................................ ................. 12 Table 7

– Studies Anticipated as Needed for the Registration Re view of HHT

........................... 14 Table 8

– Data Required by the May 12, 2010 Data Call

- In for the HHT RED

.......................... 16 Table 9

– Existing Toxicological Endpoi nts for HHT

................................ ................................ .. 17 Table 10

– Occupational Handler Exposure Scenarios for HHT

................................ .................. 19 Table 11

– Ecological Effects Endpoints Selected for Assessing Risks from HHT Use

............. 23Docket Number EPA

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## 1 Introduction

This document is the United States Environmental Protection Agency ’s ( USEPA, EPA or “the Agency ”) Preliminary Work Plan (PWP) for hexahydro

- 1,3,5

- tris (2

- hydroxyethyl)

- s

- triazi ne , also herein referred to as HHT

. The PWP 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 especially useful to the Age ncy 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

. The Agency encourages all interested st akeholders to review the PWP and to provide comments and additional information that will help the Agency ’s decision

- making process for this chemical.

The antimicrobial agent in HHT is formaldehyde (HCHO); therefore, this registration review also addresse s 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 RED is available in the formaldehyde public docket at http://www.regulations.gov in docket number EPA

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- 0121.



## 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 mu st 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 a bility to assess risk evolves and as policies and practices change, all registered pesticides continue to meet the statutory standard of no unreasonable adverse effects to human health or the environment. Changes in science, public policy, and pesticide us e practices will occur over time. Through the registration review program, the Agency periodically reevaluates pesticides to make sure that as change occurs, products in the marketplace can be used safely. Information on this program is provided at http://www2.epa.gov/pesticide

- reevaluation

.
The Agency is implementing the registration review program pursuant to Section 3(g) of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) and will review each registered pesticide every 15 years to determine whether it continues to meet the FIFRA standard for registration. The regulations governing registration review begin at 40 CF R 155.40. The Agency will consider benefits information and data as r equired by FIFRA. The public phase of registration review begins when the initial docket is opened for each case. The docket is the Agency ’s opportunity to state what it knows about the pesticide and what additional risk analyses and data or information it believes are needed to make a registration review decision. After reviewing and responding to comments and data received in the docket during this initialDocket Number EPA

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Page 8 of 58 comment period, the Agency will develop and commit to a F inal W ork P lan (FWP) and anticipated schedu le for the HHT case

. 

## 1.2 Case Overview
The docket for HHT (case #3074) has been established at http://www.regulations.gov in docket number EPA

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. Documents associated with this regis tration review can be viewed at this docket

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

.

Table 1

– Summary of Anticipated Risk Assessments and Data Needs for Re gistration 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 detai l s)

Dietary (food) No (See s ection 3.2 ) 6/30/2008 None None

Dietary (drinking water) Yes N/A New None (See section 3.2.1 )

Occupational Handler Yes 6/30/2008 Updated Immunotoxicity Indoor Inh alation Exposure

Residential Handler No (See s ection 3.3.3 ) 6/30/2008 None None

Resident i al Post Application

- Incidental Oral and Dermal No (See s ection 3.3.4 ) 6/30/2008 None None

Resident i al Post Application

- Inhalation No (See s ection 3.3.4 ) 6/30/2008 None None

Aggregate No N/A None None

Cumulative No (See section 3.4.2 ) N/A None None

Tolerance Review No (See section 1.5.1 ) N/A None None

Ecological Yes 1/7/2008 Updated Vascular plant growth Algae growth Fish early life stage Aquatic invertebrate life cycle Hydrolysis Aerobic aquatic metabolism Adsorption/desorption Waste water treatmen t 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.Docket Number EPA

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Page 9 of 58 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 Close Public Comment Pe riod 2015

- 02 Phase 2: Case Development Issue Final Work Plan 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 Clos e 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).



## 1.3 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 discuss ed in Appendix E and B, respectively

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

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Page 10 of 58 degradate 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 st udies 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 ( o C) 155 o C

830.7550 Octanol

- water partition coefficient at 25 o C (Log K ow ) - 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 (g /L) 3 g/L (EPI Suite, version 4.11)

830.7950 Vapor pres sure (mmHg) 11.2 mm Hg, at 20 C ; 15.15 mm Hg at 25 C; 61.1 mm Hg at 50 C) ; (MRID # 47284307) Mobility Organic Carbon Partitioning Coefficient Log K oc 10 L/kg (EPI Suite, version 4.11) Persistence in Water (half

- life)

Hydrolysis 1 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) Aerobic aquatic metabolism ( half

- life in days) Data ga p (See table 7 ) Adsorption/ desorption study Data gap (See table 7) Persistence in WWTP (% removed) Data gap (See table 7) [Ready biodegradation,....] 2 Data gap (See table 7) 1

. 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 limited the utility of the study. 2

. A published study ( Voet et al, 1975 ) indicated primary bi odegradation; however, this study did not follow the degradation past the first degradate.



## 1.4 Use/Usage Description

1.4.1 Registrations

There are 11 EPA

- registered products that contain HHT as an active ingredient (a.i.) and used primarily as material preservative, bactericide and fungi c ide 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, formulation intermediate, emulsifiable co ncentrate, and wettable powder. Three of these products contain other active ingredients such as sodium pyrithione or 1, 2

- benzisothiazolin

- 3

-Docket Number EPA

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Page 11 of 58 one . There is one formulation intermediate product (EPA Registration Number 1839

- 59 ) that contains 7 8.5 percent he xahydro

- 1,3,5

- tris (2

- hydroxyethyl)

- s

- triazine

.


### 1.4.2 Summary of Registered Uses

Table 5 presents a summary of the registered uses of HHT that will be assessed for registration review

. HHT can be applied by the following application methods: liquid pour, liquid pump, solid pour, and solid place. Several uses, application methods and application rates are to be mitigated as per the RED (see Table 6 )

.

Table 5

– Summary of HHT Registered Uses

Use Application Method Application Rate *

Material Preservatives

C oatings, inks, dyes and slurries Liquid Pour 1100 ppm Liquid Pump

Adhesives, dispersions, and emulsions Solid Pour 2358 ppm Solid Place Liquid Pour 1650 ppm Liquid Pump Hydrogen Sulfide Scavenger Systems Unknown 6.44 lb ai/lb H 2 S

Fuel Oils Liquid Pour 1180 ppm Liquid Pump 1180 ppm

Reagents Used in Medical Fluids Liquid Pour 393 ppm Liquid Pump 393 ppm

Metal working fluids Liquid Pou r 500 ppm Liquid Pump 500 ppm Industrial Processes and Water Systems

Coolants Liquid Pump 1180 ppm Liquid Pour

Oilfield Uses Solid Pour 1200 ppm
Solid Place 1200 ppm Liquid Pump 1200 ppm Liquid Pour 1200 ppm * Only m aximum application rates are captured

.



### 1.4.3 Summary of Risk Mitigation Measures from HHT RED

The Agency notes that there are existing labels not yet in compliance with these risk mitigation measures. The Agency is active ly taking steps to bring these labels into compliance. During

re gistration review, t he Agency does not anticipate needing to assess th o se uses that were found to be in eligible for reregistration

. 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 ).Docket Number EPA

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Page 12 of 58 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 inhalatio n 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 applicati on rate fro m 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 Recove ry 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.



### 1.4.4 Usage Information No usage information is available for HHT in the Kline Biocides Report for the period spanning 2004

- 2013

. 

## 1.5 Regulatory Histor y

The first product containing HHT as an active ingredient was regist ered in the United States in 197 2. The Agency completed a Re registration Eligibility Decision (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

.


### 1.5.1 Tolerance Information EPA has not established tolerances in raw agricul tural 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 has not been cleared as a food additive by the US F ood and Drug Administration (US FDA) under FFDCA Section 409

. 

## 1.6 Incidents

1.6.1 Human Health

In the case of HH T, there are three aggregate incidents listed in the IDS for the year 2008 , when the RED was written, to the present , based on the search conducted on O ctober 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

, andDocket Number EPA

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Page 13 of 58 burning of the lungs w ere experienced two weeks after occupational exposure to the aqueous product

.


### 1.6.2 Ecological No HHT incidents have been reported in the Agency ’s Ecological Incident Information System (EI I S) as of October 30, 2014

. 

## 2 A nticipated Data Needs

The Agency anticipates reviewing any data received 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 r egistration review

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

- RED DCI issued on May 12, 2010

.Docket Number EPA

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Page 14 of 58 Table 7

– Studies Anticipated as Needed for the Registration Review of HHT

GLN Study Name Test Substance 1 Time Frame (Me asured 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

- trimeth yl

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

835.3110 2
835.3220 2
835.3240 2
835.3280 2 Ready biodegradability

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

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

850.3300 3 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 on the degradate.

2 The results of the Activated Sludge, Respiration Inhibition Test (ASRI), GLN 850.3300, will determine which of the four biodegredation tests would be required.

 If the AS RI test results in an EC50 of less than or equal to 20 mg/L, then either the (i) Simulation tests to assess the biodegradability 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) Rea dy Biodegradability, GLN 835.3110, (ii) Simula tion tests to assess the biodegradability of chemicals discharged in wastewater, (iii) the Simulation Test

– Aerobic Sewage Treatment

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

 If the Ready Biodegradability Study is conducted and passes, th en 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 Treatm ent

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

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

- HQ

- OPPT

- 2009

- 0154

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

- ilibrary.org/content/book/9789264070080

- en

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Page 15 of 58 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 R equirement Applicable Exposure Scenario

850.1300 Aquatic invertebrate

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

850.1400 Fish early life stage TGAI 12 Ecological Assessment Industrial Wate r 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 Indust rial Water Processes, Material Preservatives Down the drain

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

870.7800 Immunotoxicity TGAI 12 All All All

875.2500 5 Inhalation Exposure Degradate: formaldehyde 24 Occupational Post

application Manufacturing Settings of

Material Preservati ves Post

- application Exposure Bystanders

__________________________ 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 guidel ine studies. See “Final Test Guidelines; OCSPP 850 Series; Notice of Availability” 77 FR 38282, June 27, 2012. http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

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- 2009

- 0154

- 0028

. 5 Formaldehyde inhalation exposures are expected to occur to worker bystanders as a result of HHT material preservative applica tions in industrial settings. Currently, no data are available to assess these worker bystander exposures

, and therefore, monitoring data are anticipated to be needed.Docket Number EPA

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Page 16 of 58 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 Ant icipated Data Requirement Applicable Exposure Scenario Status

870.1300 Acute inhalation toxicity TGAI, MP Occupational Handlers All Human Health Exposure Unsatisfied

870.3150 90

- day oral toxicity in non

- rodents TGAI Occupational Handlers; Drinking Water All Human Health Exposure Unsatisfied 1

870.3465 90

- day inhalation toxicity TGAI Occupational Handlers All Human Health Exposure Unsatisfied 1

870.3700 Prenatal developmental toxicity

study in non

- rodent TGAI Occupational Handlers; Drinking Water All Human Health Exposure Unsatisfied 1

870.3800 Reproduction and fertility effects TGAI Occupational Handlers; Drinking Water All Human Health Exposure Unsatisfied 1

870.4100 Chronic toxicity TGAI Occupational Handler; Drinking Water All Human Health Exposure Uns atisfied 1

870.4200 Carcinogenicity TGAI Occupational Handler; Drinking Water All Human Health Exposure Unsatisfied 1

875.1100 Dermal exposure

-- outdoor EP Occupational Handler Material Preservatives; Industrial Processes and Water Systems Pour Liquid Pour Solid Unsatisfied

875.1200 Dermal exposure

-- Indoor EP N/A N/A N/A Waived 2

875.1300 Inhalation exposure

-- outdoor EP Occupational Handler Material Preservatives; Industrial Processes and Water Systems Pour Liquid Pour Solid Unsatisfied

875.1400 Inhalation exposure

-- indoor EP N/A N/A N/A Waived 2

875.1600 Application exposure monitoring

data reporting EP N/A N/A N/A Waived 3

875.1700 Product Use Information EP Occupational Handler Material Preservatives; Industrial Processes and Water Systems Pour Liquid Po ur 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 Oc cupational 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 3 *N/A = not applicable 1

. Registrants ma y submit a guideline hydrolysis study (anticipated to be called

- in under registration review) to confirm the appropriate test substa nce for these toxicity studies. 2. Only outdoor exposure data is required. 3. The Agency will waive the data reporting studi es (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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## 3 Human Health Risk Assessment

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

, formal dehyde

, upon receipt of the additional data 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 an immunotoxicit y and an inhalation exposure study (as noted in Table 7 ).

The Agency is currently reevaluating the carcinogenic potential of formaldehyde and completing

a new Integrated Risk Information System ( IRIS ) assessment and unit risk valu e 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 Endp oints

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 considere d 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 d escription 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 Facto r (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 41 161801)

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

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Page 18 of 58 Exposure Scenario Dose Used in Risk Assessment (mg/kg/day) Target Margin of

Exposure (MOE), Uncertainty Facto r (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 41161 801)

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 ( M RID 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 fo r 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 mo nths) 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 than 250 mg/kg/day (the HDT); systemic LOAEL was not established.
Inh alation (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 pr ominence of the limiting ridge of the stomach in males



## 3.2 Dietary Exposure
A dietary food exposure assessment is not required for HHT at this time

. The registered uses are not expected to result in direct or indirect dietary (food) exposure

. The Agency anti cipates 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 only) assessment during Registration Review

.


### 3.2.1 Drinking Water A drinking water ass essment 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, th e magnitude of these potential residues concern, their mammalian toxicities, how persistent they will be, and 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 PWP.Docket Number EPA

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Page 19 of 58 These data will be useful to determine the extent residues of HHT and its primary triazine degradate pass through a W WTP. The Agency anticipates using these data and/or other sources of information during the ris k assessment stage of registration review to determine the potential for discharged residues of HHT to result in human dietary exposure via drinking water, which may trigger a drinking water asse ssment

. 

## 3.3 Occupational and Residential Exposures

The Agency ant icipates the need to update the occupational and residential exposure (ORE)
assessment done for the 2008 RED during registration review. The assessment from the RED will need to be re vised to account for scenarios that were identified after th e RED 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 : liquid pour, liquid pump, solid pour, and solid place

. The Agency expects to conduct new assessments for liquid pour and solid pour

scen arios in registration review upon receipt of the anticipated exposure data expected from the post

- RED DCI

.

The Agency also anticipate s that it will need to reassess the exposure of metalworking fluid and the oil field secondary recovery uses in registrati on review . The scenario may need a new risk assessment if there will be a change in the toxicological endpoint or if new exposure data are submitted in registration review. Table 10 lists the scenarios that the Agency anticipates that will be need to be as sessed in registration review.

Table 10

– Occupational Handler Exposure Scenarios for HHT

Scenario Exposure Route(s) Duration
Occupational Exposures Liquid Pour for Material Preservatives and Industrial Processes and Water Syst ems Dermal Inhalation Short and Intermediate

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

Term Occupational Handler Inhalation Exposure to Formaldehyde in Metal Working Fluids and Oil Fields Inhalation Short and Intermediate Term



### 3.3.2 Occupational Post

- application Exposures Metalworking Exposure

The RED assessed the dermal and inhalation exposure when a worker handles treated metalworking fluids with HHT and the HHT degrada te formaldehyde. According to the RED, the dermal MOE value for HHT is above the target MOE of 1,000 for intermediate

- and long

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Page 20 of 58 term exposures (MOE = 3,100). The inhalation MOE values for ST/IT/LT exposures to HHT are above the target MOE o f 100 (ST/IT/LT MOE = 44,000).

In the RED, the Agency assessed inhalation risks from HHT by using an inhalation monitoring study which measured formaldehyde levels in a metalworking facility. For the monitoring study (D346240), there were two studies presented: Linnainm aa, 2003 and Cohen, 1993 measured formaldehyde exposures from production operations. The majority of the concentrations exceeded the 100 ppb level of concern. As a result, the Agency specified reduced maximum application rates for HHT metalworking in the R ED from concentrations of 1600 ppm to 500 ppm since some samples were above the OSHA PEL ceiling value. During registration review,
t he need to revise the existing inhalation assessment for formaldehyde will be revisited once data have been received

.
Indu strial Bystander Inhalation Exposure

Formaldehyde inhalation exposures are expected to occur to bystanders as a result of HHT material preservative applications in oil fields . Currently, no data are available to assess the bystander exposures and therefore , monitoring 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 to fulfill the post

- application exposure data (Series 875.2500) in registration review.



### 3.3.3 Resident ial 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. T here are no other residential uses for HHT

. 

### 3.3.4 Residential Post

- Application Exposures

All of the HHT residential post

- application 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 risk concerns identified

. 

## 3.4 Aggregate and Cumulative Exposure

3.4.1 Aggregate Exposures

The Agency does not anticipate conduct ing an aggregate assessment as only drinking water exposures will need to be considered. R esidential an d dietary uses ( such as paint, cleaners and detergents) were identified in the RED as risks of concern and found ineligible for reregistration.



### 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 toxicityDocket Number EPA

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Page 21 of 58 with other substances. For information regarding EPA’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the c umulative 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 a quatic environment. The risk assessment also 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 as sessment 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 Environmen tal 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 ac idic pHs, shows slow hydrolysis 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 rat e of the hydrolysis reaction is 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

- trimet hyl triazine

. A second h ydrolysis 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 materia l 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 t he triazine degradate.

HHT is a volatile substance with a measured vapor pressure of 15.15 mm Hg at 25 o C and volatility is possibly due to slow release of formaldehyde from HHT

. HHT is also highly water soluble, and as water vapor volatilize s from surface water, it is likely to volatilize along with the water vapor. However, since the estimated half

- life in air is about 16 minutes, HHT is not expected to persist in air.

Although a p ublished literature study (Voets, et al , 1975 ), concluded that HHT is a rea dy 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

.Docket Number EPA

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Page 22 of 58 The e stimated K oc of HHT is 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 co mplete environmental fate database for formaldehyde, but at this time, has no environmental 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, Biod egradability) of HHT and 1,3,5

- trimethyl triazine. These studies are anticipated to 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 Cle an 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 environmental fate data presented in Table 4 and Appendix B, the Agency has determined that some exposure of a quatic 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 fluids / completion fluids ; and in

service use of in

- can preservation of adhesives, chain lubri cants, 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 i ncluded in “Conceptual Models for Environmental Exposure Pathways of Antimicrobial Pesticides” (EPA

- HQ

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

HQ

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. For industrial use of m etal 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 lubricants, 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 performs a risk assessment for HHT and it s degradates.



## 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 patterns, which indicated that rele ase

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

2 http://iaspub.epa.gov/tmdl_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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Page 23 of 58 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 determine potential acute and chronic r isks 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 com pound’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 contribute 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 a nd 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 bel ow, 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 receptor 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 11

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

, and these data are anticipated to be required

.

Table 11

– 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 9 6

- 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

-- Freshwater invertebrates Daphnia magna Acute 48

- h EC 50 = 26.1 mg/L MRID# 43175401 Chronic Data gap 2

-- Fres hwater 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

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Page 24 of 58 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 chr onic 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 Wildl ife Exposure Estimates

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 Ind ustrial 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 wastewater treatment; (2) environmental releases of antimicrobial to sur face 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 re moval of antimicrobial 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 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 = 83.8 mg/kg MRID# 43154301

Mallard duck Subacute

dietary LC 5 0 >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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Page 25 of 58 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 treatment facilities. Since HHT has the potential to be released to both d omestic 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 Analysi s Plan

Additional open literature studies will be identified through EPA’s ECOTOXicology (ECOTOX) database. The ECOTOX database will be searched during the risk assessment phase of HHT and the data reviewed following the Agency ’s policy for reviewing publi c 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 11 and / or which satisfy an existing data gap(s) w ill be used during the risk assessment phase of HHT to update Table 11 . ECOTOX was created and is maintained by the USEPA, Office of Research and Development, and the National Health and Environmental Effects Resea rch Laboratory's Mid

- Continent Ecology Division.


## 5 Endocrine Disruptor Screening Program (EDSP)

As required by FIFRA and FFDCA, EPA reviews numerous studies to assess potential adverse outcomes from exposure to chemicals. Collectively, these studies include acute, subchronic and chronic toxicity, including assessments of carcinogenicity, neurotoxicity, developmental, reproductive, and general or systemic toxicity. These studies include endpoints which may be susceptible to endocrine influence, including effec ts on endocrine target organ histopathology, organ weights, estrus cyclicity, sexual maturation, fertility, pregnancy rates, reproductive loss, and sex ratios in offspring. For ecological hazard assessments, EPA evaluates acute tests and chronic studies th at assess growth, developmental and reproductive effects in different taxonomic groups. As part of its reregistration decision for HHT , EPA reviewed these data and selected the most sensitive endpoints for relevant risk assessment scenarios from the existi ng hazard database. However, as required by FFDCA section 408(p), HHT is subject to the endocrine screening part of the Endocrine Disruptor Screening Program (EDSP).

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

a ctive and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator

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

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Page 26 of 58 may designate.” The EDSP employs a two

- tiered approach to making th e statutorily required determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal systems. Chemicals that go through Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA will determine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2 testing is designed to identify any adverse endocrine

- related effects caused by the substance, and establish a dose

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

Under FFDCA section 408(p), the Agency must screen all pesticide chemicals. Between October 2009 and February 2010, EPA issued test orders/data call

- ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. A second list of chemicals identified for EDSP screening was published on June 14, 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 po licies 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

As noted in section 1.4. 3 , the Agency is taking steps to bring outstanding HHT labels into complianc e 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 household cleaning p roduct uses need to be removed to address RED mitigation. In addition, t he Agency invites comment on any label amendments that could be considered to eliminate the anticipated need for EPA to require certain data, reduce the possibility that EPA’s planned risk assessments overestimate risk due to reliance on conservative assumptions, and/or improve label clarity.


## 7 Guidance for Commenters

7.1 Preliminary Work Plan
The public is invited to comment on EPA’s PWP and rationale. The Agency will carefully consider all comments as well as any additional information or data provided in a timely manner prior to issuing a final work plan for the HHT registration review case.



### 7.1.1 Trade Irritants Through the registration review process, the Agency intends to solicit information on trade irritants and, to the extent feasible, take steps toward facilitating irritant resolution. The Agency

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

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- 2009

- 0477

- 0074 for the final second list of che micals.

6 http://www.epa.gov/endo/Docket Number EPA

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Page 27 of 58 will work to harmonize tolerances and international maximum residue limits (MRLs) and may modify tolerance levels to do so, when possible. Stakeh olders are asked to comment on any trade irritant issues resulting from lack of MRLs or disparities between U.S. tolerances and MRLs in key export markets, providing as much specificity as possible regarding the nature of the concern.



### 7.1.2 Water Quality

The Age ncy invites submission of water quality data for this pesticide. To the extent possible, data should conform to the quality standards in Appendix A of the OPP Standard Operating Procedure: Inclusion of Impaired Water Body and Other Water Quality Data in OP P’s Registration Review Risk Assessment and Management Process 7 in order to ensure they can be used quantitatively or qualitatively in pesticide risk assessments.



### 7.1.3 Environmental Justice EPA seeks to achieve environmental justice, the fair treatment and mea ningful involvement of all people, regardless of race, color, national origin, or income, in the development, implementation, and enforcement of environmental laws, regulations, and policies. To help address potential environmental justice issues, the Agen cy seeks information on any groups or segments of the population who, as a result of their location, cultural practices, or other factors, may have atypical, unusually high exposure to HHT compared to the general population. Please comment if you are aware of any sub

- populations that may have atypical, unusually high exposure compared to the general population.



### 7.1.4 Structure Activity Relationships EPA must rely upon information of appropriate quality and reliability for each decision made by the Agency . In the OPP, the evaluation process for a pesticide chemical traditionally begins with the applicant’s submission of a set of studies conducted with the specific pesticide chemical of interest. The use of the results of such testing (measured data) is a logical, scientifically rigorous process that identifies the physical, chemical, and environmental fate properties of the pesticide, as well as the dose and endpoints at which an adverse effect can occur in various animal species.

Today, there is significant inter est in alternative techniques, i.e., techniques other than data generation that could significantly inform the Agency ’s decision

- making process. Recently, OPP has made increasing use of structure activity relationship (SAR) as part of its regulatory decisi on

- making process. In the SAR process, a chemical's molecular structure is compared to that of other chemicals for which data are available. These structural similarities are then used to make predictive judgments about a chemical’s physical, chemical, and biological properties. Thus, the chemical’s physical, chemical, and biological properties are a function of (or directly related to) the chemical’s molecular structure. Quantitative SAR is referred to as QSAR. To develop a QSAR, a selected set of measured data on a single physical, chemical, or biological property is used to derive a model (an equation) to predict the value of that property.

7 http://www.epa.gov/oppsrrd1/registration_review/water_quality_sop.htmDocket Number EPA

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Page 28 of 58 If stakeholders believe that submission of predicted data can fulfill one of the data needs for the HHT ca se, then the Agency invites submission of this information. The submitter would be expected to supply a rationale describing the utility of the information and provide documentation on the scientific validity of the information. The determination that the predicted data fulfills the data requirement would be at the sole discretion of the Agency . Pre

- submission consultation with the Agency is encouraged.



### 7.1.5 Additional Information

Stakeholders are also specifically asked to provide available information and data that w ill assist the Agency in refining its risk assessments, including any species

- specific ecological effects determinations. The Agency is interested in receiving the following information:

1. Confirmation on the following label information: A. Sites of applicati on B. Formulations C. Application methods and equipment D. Maximum application rates E. Frequency of application, application intervals and maximum number of applications F. Geographic limitations on use 2. Use or potential use distribution 3. Use history 4. Usage/use inf

ormation for non

- agricultural uses (e.g., materials preservation) 5. Typical application interval 6. State or local use restrictions 7. Ecological incidents (non

- target plant damage and avian, fish, reptilian, amphibian and mammalian mortalities) not already rep orted to the Agency 8. Monitoring data



## 8 Next Steps

After the 60

- day comment period closes in February 2015 , the Agency will review and respond to any comments received in a timely manner, and then issue a Final Work Plan for the HHT

case.Docket Number EPA

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Page 29 of 58 

## 9 References

Holtzma n, 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 Fluids,” Markuu Linnainmaa, Han nu 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, July/August 2003.

NIOSH 2003 . NIOSH Method 2016 Issue 2: Formaldehyde, NIOSH Manual of Analytical Methods (NMAM), Fourth Edition, NIOSH, March 15, 2003.

U.S. EPA, 1999. Compendium of Methods for the Determination of Toxic Organic Compounds in Ambient Air, Second Edition, Compendium M ethod TO

- 11A, Determination of Formaldehyde in Ambient Air Using Adsorbent Cartridge Followed by High Performance Liquid Chromatography (HPLC, EPA/625R

- 96/010b, EPA ORD, January 1999.

U.S. EPA, 2002. ECOTOX User Guide: ECOTOXicology Database System. Versio n 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: Degradation of Microbiocides under Different Environmental Conditions, J. Applied Bact.: 40:67

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Page 30 of 58 

# Appendix A T oxicology Profile

Acute Toxicity for Product Labeling

As listed in Table A1 , HHT wa s 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 w a 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 Categ ory 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

– Ra bbit ( 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 M ild 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 stud ies (MRID s 41 4830 02 & 41858301 ) ; all the studies are in the rat

. 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 inDocket Number EPA

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Page 31 of 58 r oden ts are anticipated 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 requir ed 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 thoraci c 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 adrenal weig hts 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 second high d ose 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 statistically

- signif icant 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 we re 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

. While this finding is not believed to be treatment

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

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Page 32 of 58 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 gastric lesio ns (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 administe red 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 water consump tion, 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) animals but wa s 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 were signif icantly 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 neurological 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 weight/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

– Ra t
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 undilu ted 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 gauze dressing covered with an Elastoplast e lastic adhesive bandage

. At the end of each 6

- hour exposure period the bandage and dressing were removed and the exposure site washed and dried.

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

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Page 33 of 58 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 fe males.

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 controls

. No intergroup difference in wate r 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 significant since they were within the normal r ange 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 h igh 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 females, and decreased chloride in mid and hig h 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 a ll 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 o rgan weights

. Treatment

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

. Histopathological findings included dose

- related epidermal ulceration, acanthosis and dermal inflammatory infiltration with occ asional 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 s ystemic 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), Vanic ide 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 divided into quadrants

. Daily applications were rota ted from quadrant to quadrant

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

. Material was left in contact with the skin for 6 hours and afterward, the residual material was removed with a paper towel and was washed wi th reverse osmosis water

. This procedure was repeated for 5 days a week forDocket Number EPA

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Page 34 of 58 the next 13 weeks

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

Three females died during the study (1 control, 1 m id 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 beca use 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 lesions , 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 animal s , 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

. Although there was no statistical significance in mea n 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 reported for females in any of the treated groups w hen 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 wa s 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 r eported 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 blood 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 chloride values were statistically lower than controls at the in terim 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 differences reported for albumen, BUN calcium, glucose, total pro tein, 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, including increased phosphorus and decreased bilirubin and cho lesterol in mid

- dose group, when compared to controls

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

- dose females

. No significant differences were reported for organ weights

. Relative increases were reported for liver and lung weights in females without statistical significance when compared to controls.Docket Number EPA

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Page 35 of 58 Histologically, there were no treatment related lesions reported with the exception of those reported for the skin

. T here was an increased incidence in hyperplasia and hyperkeratosis of t he 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, focal 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 esch ar 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 differences 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 irrit ation 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 der mal 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/day, 5 days/week for 13 weeks

. During the exposure period, the appli cation 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 ob served 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 reactions at the applications site were observed in animals from mid

- an d 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 being very severe to moderate in high

- dose animals ; whereas, only isol ated 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 in blood chemistry values from individual animals , including increa sed 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.

Microscopically, there was an increase in the incidence of epidermal hyperplasia and ulceration of the treated area of the skin in animals of the mid

- and high

- dose groups

. Eighty to 100 percentDocket Number EPA

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Page 36 of 58 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

. 

## Developmen tal 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 411 61801 & 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 registration review risk assessment

, based on the currently registered uses for HHT; this st udy was identified as a data gap in the RED

.
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, piloerection, 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 dose 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 lesions, 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 de monstrate any differences 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 addi tion, those dosage levels 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 and retarded ossification of the vertebral thoracic centra wh ic h appeared to be dose

- 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.Docket Number EPA

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Page 37 of 58 The m aternal NOAEL is 500 mg/kg/day and the LOAEL is 75 0 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 de velopmental toxicity 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

.
Mortal ity was observed 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 c ontrol animals

. 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 o bserved in the 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 com pared with the 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; s wollen hind paws 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 fetal 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 with 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 fo r the occurrence 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 <10 mg/k g/day

, the lowest 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
pr ovided to determine whether the renal observations in the pups were treatment

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Page 38 of 58 

## 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 RED and is part of the data require d 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 be 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 is part of the data required by the post

- RED DCI issued on May 12, 2010

. 

## Mutagenicity

Adequacy of D atabase for Mutageni city : 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) and found acceptable, except for a few in vitro studies

. 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 ass ay (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 th e 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 the absence and presence of a mammalian metabolic 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 increasedDocket Number EPA

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Page 39 of 58 revertent colony counts

. The exception was a significant F

- value (p < 0.05) calculated for inactivated TA1537 cultures, deri ved 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) , strains TA1535, TA100, TA102, TA 1537, TA 9 8 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 u sing 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

- incorporation and preincubation method for strains TA15 35, 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 induced the appropriate responses in the corre sponding 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 control response, thus, a new test was condu cted 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 a t 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 mutant 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 negative 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 classified as A cceptable /G uideline and satisfies the guideline require ment for Test Guideline OPPTS 870.5300, OECD 476 for in vitro mutagenicity (mammalian forward gene mutation) data

. Cytogenetics

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

- Nummer 1157) dissolved in deionized water (10% v/v) atDocket Number EPA

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Page 40 of 58 concentrations of 0, 2.5, 5, 10, 20, 30, or 40 μg/mL with and without metabolic activation (S9

- mix) for 4 hours.

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

- finding study . The range

- finding study indicated that the test substance had clastogenic effects after 4 hours ; therefore, a second experiment with a longer exposure time was no t 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; and 61, 45, and 41% of solvent controls +S9, respectively. At a concentration o f 30 μg/mL

- S9, the number of ce lls 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 evaluated 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

nu mber 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 Ch inese 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 guideline requirement for Test Guideline i n vitro mammalian cytogenetics chromosome aberration OPPTS 870.5375; O ECD 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 dose s 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

- treatment 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 effects of the test substance

. This increase

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

- Whitney test

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

. The positive contr ol induced the appropriate response

. A significant increase in the frequency of micronucleated polychromatic erythrocytes in bone marrow was not observed at either analysis time point.

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

.Docket Number EPA

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Page 41 of 58 (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/g roup) 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 approximat ely 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 selected 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 w as 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

co nsidered to be of no biological significance since it fell within solvent control ranges at other sampling times (notably 24 hours when optimally most of the micronuclei would be visualized).

Hence, the investigator concluded that triazine did not induct m icronuclei 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 Genotoxicity

(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 results of the preliminary cytotoxicity the test article

. Two UDS and parallel cytotoxicity assays were conducted wi th 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, theref

ore, 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 nuclear 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

- r elated response

. The study author concluded thatDocket Number EPA

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Page 42 of 58 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, t he study author failed to provide information on test material purity, the batch number of the assayed sample, and analytical data to verify actual concentrations used in the assay

. Therefore, it was concluded that the study is unacceptable, but the test m aterial 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 concentrations of test article (ranging from 0.001 to 0.30 μg/mL), together with a constant dose of tritiated thymi dine (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 less 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 a s 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 t o 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 proposed by the investigators, namely, increase of 5+ counts over control

. Higher doses were stated to be “too toxi c 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 concentra tions, (2) testing hepatocytes from a female rat, and (3) providing the purity (% a.i.) of the test article.

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 μg /ml.Docket Number EPA

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Page 43 of 58 Thus, these two comparable assays, when considered together, constitute the recommended confirmation of s ingularly positive results, and may be re

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

(3) In an in vivo unscheduled DNA synthesis assay in rat hepatocytes (MRID 47284304), base d 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 concentrations of 40 mg/kg and 100 mg/kg, respectively

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

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

. The positive controls elicited the appropria te responses

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

This study is classified as Acceptable/Guideline and satisf ies the guideline requirement for Test Guideline OPPTS 870.5550; OECD 4 86 for other genotoxic mutagenicity data. 

## Immunotoxicity

870.7800 Immunotoxicity Study

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

. There are no Immunotoxicity studies in the Agency ’s hazard database

. An Immunotoxicity 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 study 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 absorp tion 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 M RID 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

- thicknes s 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 (Exp eriments 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 applicationDocket Number EPA

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Page 44 of 58 (Experiments 1 and 2)

. The skin was tape stripped 8 hours after application (Experiment 2) or 24 hours follow ing 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 from 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, respectively

. The next high est 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 ap plied 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 receptor fluid and skin samples.

This in vitro study using human skin is ACCEPTABLE , NON

- GUIDELINE

. This study is considered non

- guideline because it is an in vitro human skin penetration and distribution study. This study 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 item batch #: 210405/012, 81.72% radiochemical purity; no n

- 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 chromatograms 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

. Since this impurity was greater than the impurity observed i n 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 remaining, which suggests instability in this study; therefo re, 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 uideline and does not satisfy the guideline requirement [O PPTS 870.7485, OECD 417] because the test system was not appropriate, the test material was unstable, and the study greatly deviated from the guidelines

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Page 45 of 58 Table A 2

– Toxicity Profile for HHT

Guideline No./ Study Type MRID No./

Study Classification Dosin g 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 infilt ration 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) (gavage) 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. 870.3200 21

- day Dermal (Rats) 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 inflammatory 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 (Rats) 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 hyperplasia and eschar formation).

NOAEL systemic = 30 mg/kg/day LOAEL systemic = 1 00 mg/kg/day

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

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Page 46 of 58 Guideline No./ Study Type MRID No./

Study Classification Dosin g and Animal Information Results

870.3250 90

- day Dermal Subchronic (Rats) MRID 41483002

Minimum Purity 78.5%

Rats (10/sex/dose)

0, 5, 50, 250 mg/kg NOAEL dermal = 5 mg/k g/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

. Developmental Toxicity 870.3700a Developmental Toxicity (Rat) MRID 41161801

Minimum Purit y 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

. 870.3700a Developmental 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 LOAE L 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. NOAE L 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 me tabolic 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 an d in TA1538 with and without activation.Docket Number EPA

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Page 47 of 58 Guideline No./ Study Type MRID No./

Study Classification Dosin g and Animal Information Results

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 Negative

870.5300 Mammalian cell gene mutation test MRID 47284303

Acceptable/Guideline Purity 79% aqueous solution

Chines e 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. Neg ative

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

. NegativeDocket Number EPA

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Page 48 of 58 Guideline No./ Study Type MRID No./

Study Classification Dosin g and Animal Information Results

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

Unacceptable Rat micronucleus cells

Purity 78.5%

Intraga stric intubation, dermal application, or subcutaneous injection ; 2 equal doses, separated by 24 hours, of 15, 60, 240, or 960 mg/kg Negative

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 no t 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% (r adiolabelled)

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% perce nt 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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Page 49 of 58 

# Appendix B Environmental Fate

Environment al 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 hi gh 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 raded 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 conditions, 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 stu dy was 25  ̊ C. Hydrolytic degradate characterized was formaldehyde. 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

–

p H 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 ha lf

- lives at neutral to basic pHs.

4. MRID# 48741001 : This hydrolysis 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 con ducted at 25, and 35  ̊ C. About 30% loss of released formaldehyde was noted. No rationale was provided for 30% loss of formaldehyde. Incomplete study

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Page 50 of 58 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 1/10 th 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, t he 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 active 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 t he 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 submi tted. Data gap.

Ready Biodegradability: No Data was submitted. Data gap.Docket Number EPA

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Page 51 of 58 

# 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 species 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 die tary 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 previously submitted categorize HHT as being practically nontoxic when ingested (Table 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

ora l 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 c hapter for details on the available mammalian toxicity studies

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Page 52 of 58 

### 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 pref

erred 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 ( Ic talurus 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

(Triadin e 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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Page 53 of 58 two su bmitted studies are presented in Table 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 requirement (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 exposu re levels of concern 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 24 7712

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 a pplication to the marine/estuarine environment or the active ingredient is toxic to aquatic organisms and is expected to reach this environment via other transport pathways. The preferred test species are the sheepshead minnow ( Cyprinodon variegatus ), mysi d shrimp ( Mysidopsis bahia ), and Eastern oyster ( Crassostrea virginica )

. Table 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
estu arine/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 MR ID 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 43143103Docket Number EPA

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Page 54 of 58 Aquatic Plants

No data are available. St udies 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 f our 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.Docket Number EPA

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Page 55 of 58 References

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. Unpublis hed study prepared by Wildlife International Ltd. 18 p

. MRID 43154302

Carson, S. (1968) The Effect of Grotan on Fingerling Fish: Laboratory No. 88969. (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 Flow

- Through Test Conditions: Triazine: Lab Project Number: J9306004D. Unpublished study prepared by Toxikon Envi ronmental 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 43 143103

Davis, J. (1994) Acute Effect on New Shell Growth of the Eastern Oyster, Crassostrea virginica : Triazine: Lab Project Number: J9306004H. 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 received Sep 29, 1975 under 21167

- 2; prepared by Industrial Bio

- Test Laboratories, Inc., submitted by Demille Chemical Co rp., Jersey City, N.J.; CDL: 227630

- A)

. MRID 47528

Fletcher, D. (1973) Report to Demille Chemical Corporation: 8

- Day Dietary LCI50^ Study with Milidin X

- 2 in Bobwhite Quail: IBT No. 651

- 03691. (Unpublished study received May 14, 1964 under 21167

- 2; prepare d 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 105189

LeBlanc, G.; Sousa, J. (1982) Acute Toxicity of Triadine 10 to Rainbow Trout ...: Report #BW

82

- 2

- 1117. (Unpublis hed study received Jun 11, 1982 under 125 8

- 990; prepared by EGDocket Number EPA

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Page 56 of 58 & G, Bio nomics, submitted by Olin Corp., Stamford, CT; CDL:247712

- B)

. MRID 105190

LeBlanc, G.; Sousa, J. (1982) Acute Toxicity of Triadine 10 to Bluegill ...: Bionomics Report #BW

- 82

- 2

- 1119. (U npublished 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

- 8 2

- 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 ...: Bi onomics Report #BW

- 82

- 4

- 1144. (Unpublished study received Nov 8, 1982 under 21167

- 2; prepared by EG & G, Bionomics, submitted by Demille Chemical Corp., Jersey City, NJ; CDL:248883

- B)

. MRID 118415

LeBlanc, G.; Surprenant, D. (1982) Acute Toxicity of Triadi ne

- 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 Che mical Corporation: Four

- Day Static Fish Toxicity Studies with Milidin X

- 2 in Rainbow Trout and Bluegills: IBT No. 665

- 03509. (Unpublished study 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 E nvironmental 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 t reatment 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 aquat ic 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

– Product Chemistry of HHT

Guideline Numbers HHT Source

830.1800 Enforcement analytical method HPLC 830.6302 Color Brown to dark brown 830.6303 Physical State TGAI is a solid 830.6304 Odor Aldehyde like 830.6313 Stability Stable for one year 830.6314 Oxidation/Reduction Not applic able 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 app licable 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 (EP I 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 47284307