For reasons of economy, this document is pr inted in a limited number. Delegates are kindly asked to bring their copies to mee tings and not to request additional copies.

I:\MEPC\60\2-9.doc

INTERNATIONAL MARITIME ORGANIZATION

IMO 

# E

MARINE ENVIRONMENT PROTECTION COMMITTEE 60th session Agenda item 2 MEPC 60/2/9 28 August 2009 Original: ENGLISH

HARMFUL AQUATIC ORGANI SMS IN BALLAST WATER

Application for Basic Approval of the Severn Trent DeNora BalPure ® Ballast Water Management System

Submitted by Germany

SUMMARY
Executive summary: This document contains the non-conf idential information related to the application for Basic Approva l of the Severn Trent DeNora (STDN) BalPure® Ballast Water Management System under the “Procedure for approval of ballast water management systems that make use of Active Substances (G9)” adopted by resolution MEPC.169(57)
Strategic direction: 7.1
High-level action: 7.1.2
Planned output: 7.1.2.4

Action to be taken: Paragraph 5

Related documents: BWM/CONF/36; MEPC 57/21; BW M.2/Circ.13 and BWM.2/Circ.18

Introduction

1 Regulation D-3.2 of the International C onvention for the Control and Management of Ships’ Ballast Water and Sediments stipulates that ballast water management systems that make use of Active Substances to comply with the Convention shall be approved by the Organization.

2 The “Procedure for approval of ballast water management systems that make use of Active Substances (G9)” stipulates the required information (MEPC 57/21, annex 1, paragraph 4.2.1) and provisions for risk charac terization and analysis (MEPC 57/21, annex 1, section 5), which, according to section 6 of Procedure (G9), should be evaluated by the Organization.MEPC 60/2/9 - 2 -

I:\MEPC\60\2-9.doc 3 The receiving competent authority in Germ any has verified the application dossier and believes it to satisfy the data requ irements of Procedure (G9) adopted by resolution MEPC.169(57).

4 In accordance with BWM.2/Circ.18, Germany th erefore submits the non-confidential part of the manufacturer’s application dossier in the annex. The complete dossier will be made available to the experts of the GESAMP-B WWG with the understanding of confidential
treatment.

Action requested of the Committee

5 The Committee is invited to consider the pr oposal for approval and decide as appropriate.

***MEPC 60/2/9

I:\MEPC\60\2-9.doc ANNEX

NON-CONFIDENTIAL INFO RMATION ON THE BALPURE ®
BALLAST WATER MANAGEMENT SYSTEM

Contents

1 INTRODUCTION

1.1 Overview of the BalPure® Technology

2 IDENTIFICATION OF THE SUBSTA NCE OR PREPARAT ION (G9: 4.1)

2.1 Introduction to the chemical basis of BalPure ® Ballast Water Management System 2.2 Active Substances 2.3 Relevant Chemicals 2.4 Other chemicals

3 DATA ON EFFECTS ON AQUATIC PL ANTS, INVERTEBRATES AND FISH, AND OTHER BIOTA, INCLUDING SE NSITIVE AND REPRESENTATIVE ORGANISMS (G9: 4.2.1.1)

3.1 Acute aquatic toxicity 3.2 Chronic aquatic toxicity 3.3 Endocrine disruption 3.4 Sediment toxicity 3.5 Bioavailability/biomagnification/bioconcentration 3.6 Food web/population effects

4 DATA ON MAMMALIAN TOXICITY (G9: 4.2.1.2)

4.1 Acute toxicity 4.2 Effects on skin and eye 4.3 Repeated-dose toxicity 4.4 Chronic toxicity 4.5 Developmental and reproductive toxicity 4.6 Carcinogenicity 4.7 Mutagenicity/genotoxicity 4.8 Toxicokinetics

5 DATA ON ENVIRONMENTAL FATE AND EFFECT UNDER AEROBIC AND ANAEROBIC CONDITIONS (G9: 4.2.1.3)

5.1 Modes of degradation (biotic; abiotic) 5.2 Bioaccumulation, partition coefficient, octanol/water partition coefficient 5.3 Persistence and identification of the ma in metabolites in the relevant media 5.4 Reaction with organic matter 5.5 Potential physical effects on wildlife and benthic habitats 5.6 Potential residues in seafood 5.7 Any known inter active effectsMEPC 60/2/9 ANNEX Page 2

I:\MEPC\60\2-9.doc 6 PHYSICAL AND CHEMICAL PROPERTI ES OF THE ACTIVE SUBSTANCES, RELEVANT CHEMICALS AND TREA TED BALLAST WATER (G9: 4.2.1.4)

7 ANALYTICAL METHODS AT ENVIRONMENTALLY RELEVANT CONCENTRATIONS (G9: 4.2.1.5)

7.1 Analysis of Total Residual Oxidant (TRO) 7.2 Analysis of Disinfection By-Products (DBPs)

8 USE OF ACTIVE SUBSTANCE

8.1 The manner of application

9 MATERIAL SAFETY DAT A SHEETS (G9: 4.2.7)

9.1 Active Substances hazard classification 9.2 Hazard classification of Relevant Chemicals 9.3 Hazard classification of other chemicals

10 RISK CHARACTERIZATION AND ANALYSIS

10.1 Screening for persistence, bioaccumulation and toxicity (G9: 5.1)

11 EVALUATION OF THE TREATED BALLAST WATER (G9: 5.2)

11.1 Total Residual Oxidants 11.2 Water quality parameters 11.3 Disinfection By-Products (DBP s) in treated ballast water 11.4 Toxicity testing of treated ballast water 11.5 Determination of holding time 11.6 Reaction with organic matter 11.7 Characterization of degrada tion route and rate (G9: 5.3.5) 11.8 Prediction of discharge and environmental concentrations 11.9 Assessment of potential for bioaccumulation 11.10 Effects assessment 11.11 Effects on aquatic organisms 11.12 Effects on sediment 11.13 Comparison of effect assessm ent with discharge toxicity

12 RISK ASSESSMENT

12.1 Risk to Safety of Ship 12.2 Risks to Human Health 12.3 Risks to the Aquatic Environment

13 ASSESSMENT REPORT

13.1 Overview of the Data and Endpoints

14 REFERENCESMEPC 60/2/9 ANNEX

Page 3

I:\MEPC\60\2-9.doc LIST OF TABLES

Table 1 - Overview of chemical identification Table 2 - Acute aquatic toxicity data Table 3 - Acute aquatic toxicity-treated ballast water Table 4 - Overview of chronic aquatic toxicity Table 5 - Chronic aquatic toxicity-treated ballast water Table 6 - KOCs for Active Substances and Relevant Chemicals Table 7 - BCFs for Active Substances and Relevant Chemicals Table 8 - Acute mammalian toxicity data Table 9 - Skin, eye, and dermal effects Table 10 - Repeated-dose toxicity Table 11 - Chronic mammalian toxicity Table 12 - Summary of developmen tal and reproductive toxicity data Table 13 - Summary of data on carcinogenicity Table 14 - Summary of data on mutagenicity and genotoxicity Table 15 - Summary of uptake, ab sorption, and excretion of chemicals Table 16 - Modes of degradation Table 17 - Physical and chemical properties of Active Substances Table 18 - Physical and chemical properties of Relevant Chemicals-THMs Table 19 - Physical and chemical properties of Relevant Chemicals-HAAs Table 20 - Physical and chemical propertie s of Relevant Chemicals and other chemicals Table 21 - Physical and chemical properties of treat ed ballast water Table 22 - Analytical methods Table 23 - Overview of BalPure® system alarms Table 24 - PBT criteria evaluation Table 25 - Total Residual Oxidant (TRO) measured as Total Chlorine Table 26 - Summary of water quality parameters Table 27 - Levels of Trihalomethanes (T HMs) and Haloacetic acids (HAAs) in treated fresh water Table 28 - Levels of Trihalomethanes (T HMs) and Haloacetic acids (HAAs) in treated brackish water Table 29 - Levels of Trihalomethanes (T HMs) and Haloacetic acids (HAAs) in treated
seawater Table 30 - PNEC value summary Table 31 - Summary of COPC exposure point concentrations Table 32 - Summary of dermal intake factors Table 33 - Summary of oral intake factors Table 34 - Summary of toxicity criteria Table 35 - Summary of cancer risks and non-cancer hazard s for BalPure® technicians Table 36 - Summary of cancer risks and non-cancer hazards for ship’s crew/dock workers Table 37 - Summary of cancer risks for the general public Table 38 - Summary of non-can cer hazards for general public Table 39 - Estimated concentrations of THMs in vent air and comparisons to PELsMEPC 60/2/9 ANNEX Page 4

I:\MEPC\60\2-9.doc LIST OF FIGURES

Figure 1 - Distribution of chlori ne species at different pH values Figure 2 - Distribution of bromin e species at different pH values Figure 3 - Example of chlorine demand/d ecay at three different TRO (measured as Cl 2 /L) Figure 4 - TRO concentration and effect of sodium bisulfite addition to treated water Figure 5 - Overview of the BalPure® ballasting process Figure 6 - Overview of the BalPure® de-ballasting process Figure 7 - Conceptual expos ure model, BalPure® System

LIST OF ABBREVIATIONS

Abbreviation Description Abbreviation Description

AIS Aquatic Invasive Species L/kg Liters per kilogram ATSDR Agency for Toxic Substances and Disease Registry LD50 Lethal Dose for 50% of test population AWWA American Water Works Association LEL Lower Explosive Limit

BDCM Dichlorobromomethane LOAEL Lowest Observable Adverse Effect Level BCF Bioconcentration Factor m Meters BMD Benchmark dose-response method MCA Monochloroacetic acid

Br 2 Bromine MEPC Marine Environment Protection Committee BWMS Ballast Water Management System mmHg Millimeters of Mercury
CAS Chemical Abstract Service mV Millivolts CCOHS Canada’s National Centre for Occupational Health and Safety μg/L micrograms per liter

CEM Conceptual Exposure Model μ g/ml micrograms per milliliter CFR Code of Federal Regulations (United States) μ S/cm microsiemens per centimeter

Cl - Chloride ion mA Milliamp

Cl 2 Chlorine mg/kg bw milligrams per kilogram body weight CO 2 Carbon dioxide mg/kg-d milligrams per kilogram per day CO Carbon monoxide mg/L Milligrams per liter
COPC Chemical(s) of Potential Concern MSDS Material Safety Data Sheet
CSF Cancer Slope Factor NaHSO 3 Sodium bisulfite
C x t Concentration x time NaOCl Sodium hypochlorite

DC Direct Current NIOZ Netherlands Institute for Sea Research

DBAA Dibromoacetic acid NOAEL No Observed Adverse Effect Level

DBPs Disinfection By-products NOEC No Observed Effect Concentration

DCA Dichloroacetic acid NOEL No Observed Effect Level (used interchangeably with NOAEL)MEPC 60/2/9 ANNEX

Page 5

I:\MEPC\60\2-9.doc Abbreviation Description Abbreviation Description

DOC Dissolved Organic Carbon OBr - Hypobromite ion
DPD N,N-diethyl-p-phenylenediamine OCl - Hypochlorite ion EC50 Effective concentration of substance that causes 50% of maximum response OECD Organization for Economic Co-operation and Development

EDCs Endocrine Disrupting Chemicals OIT Operator Interface Terminal EEC European Economic Community O&M Operational/Maintenance

EPI Estimation Programs Interface OSHA Occupational Safety and Health Administration ETV Environmental Technology Verification OPPTS Office of Prevention, Pesticides, and Toxic Substances FAC Free Available Chlorine ORP Oxidation Reduction Potential

FAB Free Available Bromine PBT Persistence, Bioaccumulation, Toxicity FDA Food and Drug Administration (United States) PEL Permissible Exposure Limit

FHSA Federal Hazardous Substance Act (United States) PFD Process Flow Diagram

GESAMPBWWG Joint Group of Experts on the Scientific Aspects of Marine Environmental Protection – Ballast Water Working Group pKa Dissociation Constant

H + Hydrogen ion PLC Programmable Logic Controller

H 2 Hydrogen gas PNEC Probable no Effect Concentration
H 2 O 2 Hydrogen peroxide POC Particulate Organic Carbon
HSDB Hazardous Substances Data Bank PPE Personal Protective Equipment
HSO 3 Bisulfite ion ppm Parts per million
HSO 4 Bisulfate ion PM Phase Rotation Monitor
HAA Haloacetic acid PSU Practical Salinity Units HOBr Hypobromous acid RfD Reference Dose HOCl Hypochlorous acid SIDS Screening Information Data Set IARC International Agency for Research on Cancer SCR Silicon Controlled Rectifier

IC Ion chromatography SGPT Serum Glutamate-Pyruvate Transaminase ILCR Incremental Lifetime Cancer Risk STDN Severn Trent De Nora

IMDG International Maritime Dangerous Goods Code TCA Trichloroacetic acid

IMO International Maritime Organization THMs Trihalomethanes

ICSC International Chemical Safety Card TOC Total Organic Carbon

IPCS International Programme on Chemical Safety TOXNET Toxicology Data Network

IRIS Integrated Risk Information System TRO Total Residual OxidantMEPC 60/2/9 ANNEX Page 6

I:\MEPC\60\2-9.doc Abbreviation Description Abbreviation Description

IUPAC International Union of Pure and Applied Chemistry TSS Total Suspended Solids

Koc Organic Carbon Partition Coefficient UCL Upper Confidence Level

Kow Octanol/Water Partition Coefficient US EPA United States Environmental Protection Agency Kpa Kilopascal vdc Volts of continuing current L/min Liters per minute WHO World Health Organization wt % Weight PercentMEPC 60/2/9 ANNEX

Page 7

I:\MEPC\60\2-9.doc 1 INTRODUCTION

Severn Trent De Nora (STDN), LLC, www.severntr entdenora.com, is a joint venture subsidiary of Severn Trent Services and Gr uppo De Nora. STDN is the world l eader in the supply of on-site generation of biocide solutions to the power, marine, offshor e industrial water disinfection markets. Over the past 30 years, STDN organi zations have supplied 65% of the worldwide installed base of on-site generated hypochlorit e. STDN draws upon th e strength and global resources of Severn Trent Services and Gruppo De Nora. The joint venture was formed in 2001 by the combination of their respective subsid iaries, Exceltec International Corporation and DeNora SeaClor, S.r.l. The history of STDN includes: 1974 – Diamond Shamrock Corporation, Electrolytic Sy stems Division formed; 1986 – Acquisition of OMNIPURE by Diamond Shamrock Corporation; 1987 – Formati on of Eltech International Corporation (Sugar Land, TX); 1997 – Eltech International Corpora tion name changed to Exceltec International Corporation; 1998 – Exceltec International Corp oration acquired by Severn Trent Services; 2001 – Severn Trent Services enters into joint venture with Gruppo De Nora; and 2002 – Severn Trent De Nora is formed. Established in 1923 by Or onzio De Nora, Gruppo De Nora is recognized worldwide as a leading supplier of electrochemical technologies for the production of chlorine, caustic soda and derivatives, as well as the largest worldwide supplier of noble metal-coated electrodes for the chlor-alkali industry and for th e electrochemical industry in general. Energy saving and environmental protection are the Group's distinguishing t echnologies. Gruppo De Nora has built more than 500 plants for the pr oduction of chlorine, caustic soda and for the electrochlorination of industrial waters in 60 different countries.

The BalPure ® system and the testing for the installation on board the S/R American Progress has been acknowledged/approved by the states of Fl orida, Texas, and Louisiana (United States). Additionally, all requirements for acceptance of the BalPure ® system on the S/R American Progress into the United States Coast Gu ard Shipboard Technology Evaluation Program (STEP) have been completed, with issu ance of the acceptance certi ficate anticipated by the third quarter of 2009.

STDN is submitting this dossier to obtain the International Maritime Organization’s Basic Approval under Procedure (G9) (M EPC 57/21, resolution MEPC.169(57)) for the BalPure ® Ballast Water Management System (BWMS).

1.1 Overview of the BalPure ® technology

The BalPure ® system provides a safe and economic elect rolytic process for th e on-site generation of a biocide solution from seawater to disinfect aquatic invasive species (AIS) in ballast water. Ballast water treatment is accomplished using a simp le three-step process of filtration, injection of a biocide solution, and residua l oxidant neutralization. The first phase is filtration using a 40 micron stainless steel mesh filter to remove large particles and sediment. The second phase of the treatment process is electrochemical gene ration of the biocide solu tion. This involves passing a small supply (1/100 of total ballast flow) of seawater, either from the incoming ballast water line or sea cooling water, through electrolytic cells. The re sulting disinfecta nt solution is injected directly into the incoming ballast wate r line where it will oxidize potential AIS. The third and final treatment process phase is residual oxidant neutralization to ensure environmental acceptability. When the treated ballast water is ready to be discharged, sodium bisulfite is injected directly into the ballast water discharge line.MEPC 60/2/9 ANNEX Page 8

I:\MEPC\60\2-9.doc The BalPure ® system is designed to require minimal i nput from the ship’s crew, with the system starting and stopping the necessary treatment steps based on electr onic signals. This minimizes added duties to ship’s crew and ensures proper ballast water treatment.

STDN’s experience with electrochlorination in a variety of industrial and maritime settings has proven the effectiveness of the technology. As discussed further in section 2.2.1, the concentration of the biocide solution needed for e ffective ballast water treatment is related to the total organic carbon (TOC) present in th e water. Efficacy of the BalPure ® system has been tested in both pilot-scale and full-scale scenarios. A pilot-scale efficacy study was conducted by the University of Washington. Results indicat ed that a residual chlorine concentration of 2.95 mg/L-3.71 mg/L with ballast seawater containing <2 mg/L TOC was sufficient to inactivate 98%–100% mesozooplankton, 99% –99.9% phytoplankton and 99%–100% bacteria in 5 hours (Herwig et al. , 2006). Within 24 hours of treatment, the residual chlorine was neutralized with the addition of sodium bisulfite . Testing of the treated water was shown to be non-toxic to aquatic organisms following th e sodium bisulfite neutralization step.

A full-scale study was conducted at the Naval Research Laboratory (NRL) in Key West, Florida (United States). The tests were part of the Environmental Tec hnology Verification (ETV) Program instituted by the United States Environmental Protection Agency (US EPA) to verify environmental treatment performance as well as operational performance. The test protocol evaluated biological treatment performance ba sed on IMO Guidelines (G8) and ETV test criteria, operation and maintenance, reliability, cost factors, environmental acceptability, and safety. Examination of these factors was accomplished thr ough two types of tests: Operational/maintenance (O&M) tests and chal lenge water tests. Based on the test results, it was found that a residu al chlorine concentratio n of 14 mg/L-16 mg/L tr eating ballast seawater containing 10 mg/L TOC effectively inactivated >99% zooplankton, phyt oplankton and bacteria after a 24-hour treatment period (NRL, 2007).

During 2009, efficacy of the BalPure ® system will be tested accordi ng to IMO Guidelines (G8) at the Royal Netherlands Institute for Sea Resear ch (NIOZ) land-based te sting facility under the direction of Dr. Marcel Veldhuis. La nd-based testing of a commercial BalPure ® system will also be conducted at the Maritime Environmental Resource Center in Maryland, USA during the fall of 2009.

2 IDENTIFICATION OF THE SUBSTA NCE OR PREPARAT ION (G9: 4.1)

2.1 Introduction to the chemical basis of BalPure ® Ballast Water Management System

Chlorine, often in the form of sodium hypochlor ite, has been used for many years in a wide variety of water treatment applications. A few examples include drinking/potable water disinfection, biofouling contro l, recreational water (swimmi ng pools/spa) treatment, and wastewater treatment. When chlorine is used fo r water treatment, hypochlorous acid is instantly produced over the pH 4-8 range. Therefore, regardless of what form of chlorine (gaseous or sodium hypochlorite) is being used to treat ball ast water, effective disinfection is largely the result of the properties of hypochlorous acid.

The BalPure ® system uses electrolysis of seawater to safely generate sodium hypochlorite on board, and immediately injects the solution directly into ballast water while it is pumped on board. Seawater is passed throug h electrolytic cells (electrolyzer s) where it is subjected to medium amperage, low voltage direct current. Electrolytic cells are co mprised of a cathode and an anode. At the outlet of the electrochemical generator, the s eawater contains a mixture ofMEPC 60/2/9 ANNEX

Page 9

I:\MEPC\60\2-9.doc chemicals produced at both the anode and cathode. Because seawater contains bromide ions, the electrochemical process results in the formation of hypochlorous acid (HOCl) in equilibrium with hypochlorite ion (OCl - ), hyprobromous acid (HOBr) in equi librium with hypobromite ion (OBr - ), and hydrogen gas (H 2 ) produced as a by-product . During the electrochemical process a small amount of caustic (OH - ) is also produced, which is effectively neutralized within the cell and causes no apparent increase in the pH of treated ballast water.

Further, chlorine and bromine can undergo reactions with ammonia naturally present in seawater and may result in the formation of low concen trations of chloramines and bromamines. Chloramine and bromamine are important to di scuss in disinfection chemistry because these species are also considered disinfectants. Ho wever, they are less effective disinfectants (i.e. weak oxidants) and the amount of disinfecti on attributed to chlora mine and bromamine is not considered relevant. The amount of chlora mine and/or bromamine potentially formed is related to the amount of ammonia present, amount of oxidant pres ent, reaction time, and pH of the water treated. The amount of chloramine a nd bromamine formed during use of the BalPure ® system is expected to be insignificant because of the neutral pH at which treatment is occurring and the long reaction time required for their formation. In treated discharge, chloramines and bromamines have not been determined analytic ally for several reasons. First, the methods capable of measuring combined halogen species are not readily available, thereby making individual quantification of chloramine and brom amine difficult. Second, combined chlorine and bromine species are included in the TRO measurement. Lastly, the BalPure ® system applies a dechlorination/dechloamination pro cess upon discharge of the treated ballast water. The use of sodium bisulfite or sodium metabisulfite is an effective means of removal for any combined chlorine or bromine species acco rding to the following reactions:

NaHSO 3 + NH 2 Cl + H 2 O → NaHSO 4 + NH 4 Cl Na 2 S 2 O 5 + NH 2 Cl → 2NaHSO 4 + NH 4 Cl

The measurable oxidant species present durin g chlorine-based wate r disinfection, and the terminology used, is often a point of confusi on. By industry standards and convention, all chlorine present in a water sample, regardless of form, is referred to as Free Available Chlorine (FAC). FAC includes Cl 2 , HOCl, and OCl - . When including chlorami nes, the term Combined (or Total) Chlorine is used. Similarly, all bromine species present in a water sample are referred to as Free Available Bromine (FAB). When including br omamine, the term us ed is Combined (or Total) Bromine. The common fact or in all of these chemical spec ies is that they are considered oxidants, and due to the difficu lty in measuring each chemical species individually in a complex water sample, the term Tota l Residual Oxidants (TRO) is used to include all of these oxid ants when present in a sample. To clarify:

Free Available Chlorine (FAC) = Cl 2 + HOCl + OCl Combine (or Total) Chlorine = FAC + chloramine Free Available Bromine (FAB) = Br 2 + HOBr + OBr Combine (or Total) Bro mine = FAB + bromamine Total Residual Oxidants (TRO) = FA C + chloramine + FAB + bromamine

Therefore, in this dossier, TRO is used to describe the total of all oxidants present in treated water. The chemical methods used for analysing TRO of ten state that the measurement is calculated as mg Cl 2 /L. However, these methods include all of the oxidants present in a water sample as mentioned above, not just Cl 2
.MEPC 60/2/9 ANNEX Page 10

I:\MEPC\60\2-9.doc Using the definitions of Active Substances, Relevant Chemicals, and other components as provided in the Methodology for information gathering and the conduct of work of the GESAMP-BWWG (23 May 2008), Severn Trent De Nora classifies the substances associated with the BalPure ® system as follows:

Active Substances : As described above, the electrochemical process generates a mixture of chemicals. Of these, hypochlorous acid (HOCl) in equilibriu m with hypochlorite ion (OCl-) and hyprobromous acid (HOBr) in equ ilibrium with hypobromite ion (OBr-) are the actual disinfecting agents. These Activ e Substances are disc ussed in detail in section 2.2. In this dossier, these Active Substances are considered together and referenced as Total Residual Oxidants (TRO).

Relevant Chemicals : Disinfection of water with chlori ne-based disinfectants can result in the formation of trihalomethanes (THMs) , haloacetic acids (HAAs), and bromate that may be present at detectable levels in treated ballast water. THMs and HAAs are discussed as Relevant Chemicals in s ection 2.3.1, while bromate is discussed in section 2.3.2.

During the electrolysis process, hydr ogen gas is produced. For this reason, H 2 gas is classified as a Relevant Ch emical, however, the hydrogen gas produced is immediately separated from the solution and removed fr om the system via ventilation to the atmosphere. Though it is not present in treated ballast water, but need s to be considered when evaluating the BalPure ® system, hydrogen gas is considered a Relevant Chemical and is discussed in more detail in section 2.3.3.

Other chemicals (i.e. Other components) : Sodium bisulfite is used with the BalPure ® system to neutralize residual oxidants that may be present in treated ballast water discharge. Because sodium bisulfite is not an Active Substance, and is not produced during or after employment of the treatment sy stem (Relevant Chemical), it is considered to fall under the ‘other chemicals’ category. S odium bisulfite is discussed in more detail in section 2.4.1.

Table 1 below provides a summary of the s ubstances associated with the BalPure ® treatment system. Table 1 : Overview of Chemical Identification

Ingredient (IUPAC Name) Identification Numbers Chemical Formula Structural Formula Molecular Weight Percentage (by weight)

Active Substances in Ballast Water Hypochlorous acid CAS # 7681- 52-9 HOCl 52.46 <25 mg/L TRO Hypobromous acid CAS # 13824-96-9 HOBr 96.91 <25 mg/L TRO Relevant Chemicals
Bromate (as sodium bromate) CAS #7789- 38-0 NaBrO 3 150.89 By-product

Hydrogen CAS #133- 74-0 H 2 H---H 2.016 By-productMEPC 60/2/9 ANNEX Page 11

I:\MEPC\60\2-9.doc Ingredient (IUPAC Name) Identification Numbers Chemical Formula Structural Formula Molecular Weight Percentage (by weight)

THMs
Chlorodibromomet hane CAS #124- 48-1 CHBr 2 Cl 208.29 By-product

Dichlorobromomet hane CAS #75-27- 4 CHBrCl 2 163.8 By-product

Tribromomethane (bromoform) CAS #75-25- 2 CHBr 3 252.77 By-product

Trichloromethane (chloroform) CAS # 67- 66-3 CHCl 3 119.38 By-product

HAAs
Bromochloroacetic acid CAS # 5589- 96-8 C 2 H 2 BrCl O 2 173.39 By-product

Monochloroacetic acid CAS #79-11- 8 C 2 H 3 ClO 2 94.50 By-product

Dibromoacetic acid CAS #631- 64-1 Br2CHCO OH 217.84 By-product

Dichloroacetic acid CAS # 79- 43-6 CHCl 2 CO OH 128.9 By-product

Trichloroacetic acid CAS # 76- 03-9 CCl 3 COO H 163.4 By-product

Monobromoacetic acid CAS # 79- 08-3 C 2 H 3 BrO 2 138.95 By-product

Other Chemicals

Sodium Bisulfite CAS # 7631- 90-5 NaHSO 3 104.06 Neutralizing agentMEPC 60/2/9 ANNEX Page 12

I:\MEPC\60\2-9.doc 2.2 Active Substances

2.2.1 Hypochlorous acid and hypobromous acid

To avoid storing Active Substances on board, the BalPure ® system safely generates sodium hypochlorite by electrolysis of se awater. Seawater is passe d through electrolytic cells (electrolyzers) where it is s ubjected to medium amperage, low voltage direct current. Electrolytic cells are comprised of a cathode and an anode. The primary cell reactions occurring in the electrochemical generation pr ocess in seawater are as follows:

Reaction 1 (at the cathode): 2H 2 O + 2e - → H 2 + 2OH Reaction 2 (at the anode): 2Cl - + 2H 2 O → Cl 2 + 2e -

------------- --------------- ----------- Reaction 3 (overa ll reaction): 2Cl - + 2H 2 O → Cl 2 + H 2 + 2OH When the chlorine is produced, it is ra pidly hydrolyzed and fo rms hypochlorous acid:

Reaction 4: Cl 2 + H 2 O ↔ HOCl + H + + Cl Hypochlorous acid (HOCl) undergoes a pH-depende nt dissociation to form hypochlorite ion (OCl - ):

Reaction 5: HOCl ↔ OCl - + H +

Thus, depending on pH, one observes a distribution of chlorine sp ecies as shown in figure 1 below.

Figure 1 : Distribution of Chlorine Species at Different pH Values

As discussed in section 2.1, the combination of Cl 2 , HOCl, and OCl - are commonly referred to as free available chlorine (FAC). As figure 1 de picts, at the pH of s eawater (typically ranges from 7.5-8.4), the solution generated by the BalPure ® system will consist of a near equal mixture of hypochlorous acid (HOCl) and hypochlorite ion (OCl - ).MEPC 60/2/9 ANNEX Page 13

I:\MEPC\60\2-9.doc Because seawater also contains br omide ion, it is important to note that reactions similar to those occurring with chloride ion can al so be occurring with bromide ion in the electrolytic cell. For example, bromide ion will form Br 2 :

Reaction 6 (at the anode): 2Br - + 2H 2 O → Br 2 + 2e When bromine is produced, it is rapidl y hydrolyzed and forms hypobromous acid:

Reaction 7: Br 2 + H 2 O ↔ HOBr + H + + Br Hypobromous acid (HOBr) undergoe s a pH-dependent dissociation to form hypobromite ion (OBr - ):

Reaction 8: HOBr ↔ OBr - + H +

Thus, depending on pH, one observes a distribution of bromine sp ecies as shown in figure 2 below.

Figure 2 : Distribution of Bromine Species at Different pH Values

As figure 2 depicts, at the pH of seaw ater, the solution generated by the BalPure ® system will also consist of a mixture of hypobro mous acid (HOBr) and hypobromite ion (OBr - ), with a slightly greater proportion of the mixture existi ng as HOBr. As discussed in section 2.1, the combination of Br 2 , HOBr, and OBr - are commonly referred to as fr ee available bromine (FAB).

When using chlorine-based water disinfection, it is important to discuss the concept of oxidant demand and decay. Oxidant demand is defined si mply as the difference between the amount of oxidant added to the water and th e amount of oxidant re maining at the end of a specified contact period (i.e. total residual oxidant (TRO)). When an oxidant is added to natural waters, it will react (oxidize) with organisms and other components (e.g., natural minerals, organic matter) normally present in natural waters. Th ese components, often measured as total organi c carbon (TOC), determine the oxidant demand of the water. As su ch, for effective treatment, an oxidant needs to be applied at a concentration that exceeds the demand of the water source and also maintain a residual concentration within the ballast tanks. This allows for sufficient concentration and contact time (C x t) to ensure complete and effective inactivation of AIS.MEPC 60/2/9 ANNEX Page 14

I:\MEPC\60\2-9.doc Regardless of the initial oxidant dose, the resi dual oxidant concentration will decay over time. Studies on the demand/decay of TRO (measured as mg Cl 2 /L) produced by the BalPure ® system have been performed by the University of Wa shington. Figure 3 provides an example of the decay of TRO observed over time for varying concentrations of hypochlorite treatment.

Figure 3 : Example of Chlorine Demand/Decay at three different TRO (measured as Cl 2 /L) treatments over time . (Treatment 4.A, 3.6 mg/L; Treatment 4.B, 3.6 mg/L with immedi ate neutralization; Treatment 4.C, 6.0 mg/L) (Herwig, et al. , 2006).

Because water qualities where ballast water upta ke may occur can vary by location and season, this can impact the oxidant demand. This in tu rn can affect the hypochl orite dose needed to adequately inactivate AIS. Therefore, the e ffective concentration generated by the BalPure ® system is adjusted based on the TOC concentra tion. Low TOC concentra tions require a lower hypochlorite dose, while higher TOC concentrations require a higher hypochlorite dose to meet the oxidant demand of the incoming ballast water and also maintain a residual concentration. Empirical studies with pilot and full-scale systems over a wide ra nge of water qualities and TOC levels have shown that an appl ied concentration of <20 mg/L of hypochlorite solution results in effective ballast water treatment. This relations hip can be seen in the brief summary of the efficacy results presented above in section 1.1 re garding testing conducted at the University of Washington and the Naval Re search Laboratory.

The BalPure ® system is designed to control the am ount of Active Substances produced to effectively treat AIS, yet avoid over-production of the biocide to control the level of potential DBPs. This is accomplished using oxidati on-reduction potential (O RP) technology. Although ORP technology does not directly measure the amount of Active Substances present in the water the way a chlorine residual measurement would, OR P allows for an in-line rapid determination of the amount of oxidation (or reduction) potential of the water. ORP is measured in millivolts (mV) and increases or decrease s in direct relation with the oxidant residual in the water (Kelley, 2004). For example, natural waters havean ORP reading in th e range of 100 mV–150 mV. When hypochlorite is added to water, the ORP read ing increases with an increase in hypochlorite dosage. Similarly, when a de-chlorinating agent such as sodium bisulfite is added to the water, the ORP reading will drop to a low mV level (K elley, 2004). In this way, ORP measurements provide a practical and efficient method of c ontrolling biocide dosage and ensure optimized water disinfection. More disc ussion of how the BalPure ® system incorporates ORP technology is presented in section 8.1.1.MEPC 60/2/9 ANNEX Page 15

I:\MEPC\60\2-9.doc 2.3 Relevant Chemicals

2.3.1 Trihalomethanes (THMs) and Haloacetic Acids (HAAs)

During the disinfection process, re actions with organic materials present in water may result in the formation of THMs and HAAs. The amount of THMs and/or HAAs formed as disinfection by-products (DBPs) will vary depending on water quality. During pilot scale testing of the BalPure ® system at the University of Washingt on, the following THMs have been measured in treated ballast water: chloroform, dich lorobromomethane, chlorodibromomethane, and bromoform. The following HAAs have b een measured in treated ballast water: bromochloroacetic acid, monochloroacetic acid, dichloroacetic acid, trichloroacetic acid, monobromoacetic acid, and dibromo acetic acid. The presence of tribromoacetic acid was not determined analytically during pilot studies and is not anticipated to be present at environmentally relevant concentrations. Howe ver, tribromoacetic acid will be evaluated during land-based testing and reported in a subsequent a pplication for Procedure (G9) Final Approval. The existing data for Relevant Chemicals in treated ballast water is presented in section 11, “Evaluation of Treated Ballast Water”.

2.3.2 Bromate

Formation of bromate may occur during the electr ochemical treatment process. The following equation shows the reaction o ccurring at the electrode:

Reaction 9: Br - + 3H 2 O → BrO 3 - + 6e The amount of bromate formed will vary based on the amount of bromide ion present in the incoming seawater and the overall production rate of the treatment system. The concentration of bromate in brackish water and seaw ater treated with the BalPure ® system has been analysed and this data is presented in Tables 28 and 29, respec tively. This constituent will be evaluated further during land-based testing.

Other potential mechanisms for bromate ion form ation include the seconda ry reaction of HOCl with bromide ion to form hypobromite ion, whic h can further react to form bromate ion.

Reaction 10: HOCl + Br - → HOBr + Cl Once formed either by the generation of bromine at the anode or from a subsequent secondary reaction, hypobromous acid (HOBr) dissociates and depending on the pH of the media, will react to form a low level intermedia te species (bromite ion, BrO 2 - ), and eventually bromate ion (BrO 3 - ). The reactions of this secondar y pathway of bromate formation are:

Reaction 11: HOBr ↔ OBr - + H + Reaction 12: OBr - + OBr - → BrO 2 - + Br - Reaction 13: BrO 2 - + OBr - → BrO 3 - + Br As mentioned previously, reactions 10-13 are dependent on the concen tration of available bromide ion and pH. At the conc entrations of oxidant produced in the electrolytic cell, the formation of bromate ion through these secondary pathways is expected to be minimal.MEPC 60/2/9 ANNEX Page 16

I:\MEPC\60\2-9.doc 2.3.3 Hydrogen gas

Hydrogen (H 2 ) is formed as a by-product of seawater el ectrolysis, as can be seen in Reaction 3 above. Due to the explosive properties of hydrogen gas, allowing it to accu mulate in the ballast tanks would create an unsaf e situation. Therefore, th e hydrogen produced during the electrochlorination process is separated immedi ately upon exit from the electrolytic cell using a patented cyclone separation de vice and is not allowed to enter the ballast water piping. The separated hydrogen gas is diluted to less than 1% hydrogen by forced air blowers and vented to the atmosphere outside of the ship. The lower explosive limit (LEL) for hydrogen is 4% and the operating limitations of the system pr ovide a four-fold safety factor.

As an atmospheric gas that will not be present in treated ballast discharge it is not evaluated in regards to aquatic toxicity. Fu rther, exposure to atmospheric ga ses typically are not included in risk assessment, and the safety f eatures inherent in the BalPure ® BWMS as well as the physical properties of hydrogen gas suggest it is unlikely to accumulate to levels of concern (e.g., above the LEL). Therefore, hydrogen gas is not consider ed relevant for the assessment of human health risks.

2.4 Other chemicals

2.4.1 Sodium bisulfite

During discharge of treated water (de- ballasting), sodium bisulfite (NaHSO 3 ) is injected into the ballast water discharge line to ne utralize residual oxidants that may be present. Bisulfite addition is monitored and controlled to apply a small exce ss (<2 mg/L) to ensure that no residual oxidants (HOCl/HOBr) are present in the effluent water. The bisulfite chemically reduces any free chlorine and/or hypochlorous acid to chloride ion, the bromine and/or hypobromous acid to bromide ion.

Reaction 14: HSO 3 - + Cl 2 + H 2 O → HSO 4 - + 2HCl Reaction 15: HSO 3 - + Br 2 + H 2 O → HSO 4 - + 2HBr

Because the pKa for the bisulfat e ion/sulfate ion equilibrium is approximately 2, any residual bisulfate ion is expected to be in the form of sulfate ion and not cont ribute to a measurable change in the residual sulfate ion concentration.

Reaction 16: HSO 4 - ↔ SO 4 2- + H +

Seawater typically contains approximately 3 g/L (or 3,000 mg/L) of sulfate ion. On a theoretical basis the reaction of the residual oxidant will require 1.77 parts of sodium bisulfite by weight to remove one part of halogen (prese nt at mg/L concentrations). T hus, the concentration of sodium bisulfite that might be added to the discharged ballast water is negligible and by definition the increased sulfate ion concentration in the seawat er will also be negligible. The reaction might produce up to 5 mg/L of acid, which in the hi ghly buffered ballast water will not lower the apparent pH of the treated water.MEPC 60/2/9 ANNEX Page 17

I:\MEPC\60\2-9.doc Sodium bisulfite is a very effective oxidant neutralizing agent and an example of TRO concentrations after addition to water treated with the BalPure ® system is presented in figure 4.

Figure 4 : TRO concentration and effect of so dium bisulfite addition to treated water (Herwig et al. , 2006)

Because sodium bisulfite is added to treated ballast wate r discharge in slight excess, Severn Trent has evaluated the aquatic toxicity to confirm th at the discharge is environmentally acceptable. Toxicity data is presented in Tables 2 thr ough 5 and discussed further in section 11.4.

3 DATA ON EFFECTS ON AQUATIC PLANTS; INVERTEBRATES AND FISH, AND OTHER BIOTA, INCLUDING SE NSITIVE AND REPRESENTATIVE ORGANISMS (G9: 4.2.1.1)

A literature search was conducted to locate exis ting aquatic toxicity da ta for the substances associated with the BalPure ® system. The data summarized in Tables 2 and 4 present the acute and chronic toxicity data that wa s identified. A complete base-set for fish, crustaceans, and algae was not located for all substances and in some cases, the data do not meet validity criteria. However, the data has been provided as suppor ting information and is discussed in relation to PNEC derivation in section 11.11.

Aquatic toxicity data on whole effluent from treated ballast water studies of the BalPure ® system is presented in Tables 3 and 5.MEPC 60/2/9 ANNEX Page 18

I:\MEPC\60\2-9.doc 3.1 Acute aquatic toxicity

Table 2 : Acute Aquatic Toxicity Data

Test Substance F C

A Species Test Type E(L)C50 mg/L NOEC mg/L Reference/ comments

F Oncorhynchus mykiss 15-d survival 0.059 N/A Fisher et al. (1999)

C Daphnia magna 48-h survival 0.032 N/A Fisher et al. (1999) TRO (Freshwater) A Natural phytoplankton 47-h change in Chl a <0.1 N/A Brooks and Liptak (1978)

F Menidia beryllina 96-h survival 0.143 N/A Fisher et al. (1999)

C Mysidopsis bahia 96-h survival 0.062 N/A Fisher et al. (1999) TRO (Saltwater)

A Dunaliella primolecta 24-h growth 0.075 N/A Videau et al. (1979)

Active Substance

F Oncorhynchus mykiss 96-h survival 0.032 N/A Thatcher (1978) (Taken from US EPA ECOTOX database)

C Americanmysis bahia 7-d survival 0.144 N/A Fisher et al. (1994) Hypochlorous acid

A Porphyra yezoensis 10-d growth 1.4 N/A Maruyama et al. (1988) Hypobromous acid No Data found in literature Relevant Chemicals

F Morone saxatilis 96-h survival 31 N/A Richardson et al. (1981)

C Neomysis awatschensis 24-h survival 176 N/A Crecelius (1979) (Taken from US EPA ECOTOX database) Bromate

A Glenodinium halli Not specified 13.6 N/A Hutchinson, et al. (1997) THMs

F Lepomis macrochirus 96-h survival 14 N/A Anderson & Lusty (1979) (Taken from US EPA ECOTOX database) Chloroform

C Artemia salina 24-h survival 30 N/A Foster and Tullis (1984) (Taken from US EPA ECOTOX database)MEPC 60/2/9 ANNEX Page 19

I:\MEPC\60\2-9.doc Test Substance F C

A Species Test Type E(L)C50 mg/L NOEC mg/L Reference/ comments

A Scenedesmus subspicatus 48-h growth 950 N/A Kuhn and Pattard (1990) (Taken from US EPA ECOTOX database)

F Cyprinodon variegates 96-h survival 18 2.9 Heitmueller et al. (1981) (Taken from US EPA ECOTOX database)

C Daphnia magna 48-h immobilization 46 N/A LeBlanc, G.A. (1980) (Taken from US EPA ECOTOX database) Bromoform

A Skeletonema

costatum 96-h growth inhibition 12.3 N/A US EPA (1978) (Taken from US EPA ECOTOX database)

Chlorodibromomethane F Cyprinus carpio 72-120-h egg fertilization to hatch 34 N/A Mattice et al. (1981)

Dichlorobromomethane F Oryzias latipes 96 h survival 72 N/A Toussaint, et al. (2000) HAAs

Dichloroacetic acid C Daphnia magna 24-h immobilization 106 N/A Trenel and Kuhn (1982) (Taken from US EPA ECOTOX database)

F Pimephales promelas 96-h survival 2000 N/A Dennis et al. (1979) (Taken from US EPA ECOTOX database)

C Daphnia magna 48-h immobilization 2000 N/A Dennis et al. (1979) (Taken from US EPA ECOTOX database) Trichloroacetic acid

A Chlorella pyrenoidosa 72-h growth N/A 500 Huang and Gloyna (1968) (Taken from US EPA ECOTOX databases)

Dibromo-acetic acid F Pimephales promelas 24-h survival 160 N/A Mayes (1985) (Taken from US EPA ECOTOX database)MEPC 60/2/9 ANNEX Page 20

I:\MEPC\60\2-9.doc Test Substance F C

A Species Test Type E(L)C50 mg/L NOEC mg/L Reference/ comments

F Pimephales promelas 48-h survival 107 N/A Mayes (1985) (Taken from US EPA ECOTOX database)

F Pimephales promelas 72-h survival 81 N/A Mayes (1985) (Taken from US EPA ECOTOX database)

F Pimephales promelas 96-h survival 69 N/A Mayes (1985) (Taken from US EPA ECOTOX database)Bromochloroacetic acid No data found in literature

F Cyprinus carpio 5-h survival 222 N/A Loeb & Kelly (1963) (Taken from US EPA Ecotox Databases)

C Daphnia magna 24-h survival 34 N/A Kuhn (1988) (Taken from US EPA Ecotox Databases)Monobromoacetic acid

A Scenedesmus subspicatus 48-h survival 0.34 N/A Kuhn & Pattard (1990) (Taken from US EPA Ecotox Databases)

F No data found in literature

C Daphnia magna 48-h mobility 96 N/A Kuhn & Pattard 1990 (Taken from US EPA Ecotox Databases)

A Scenedesmus subspicatus 48-h growth 0.028 N/A Kuhn & Pattard (1990) (Taken from US EPA Ecotox Databases)Monochloroacetic acid

A Scenedesmus subspicatus 48-h growth 0.07 N/A Kuhn & Pattard, (1990) (Taken from US EPA Ecotox Databases) Other Chemicals

Sodium bisulfite F Carassius gibelio 96-h survival 100 N/A MSDS Solvay Chemicals (2007)MEPC 60/2/9 ANNEX Page 21

I:\MEPC\60\2-9.doc Test Substance F C

A Species Test Type E(L)C50 mg/L NOEC mg/L Reference/ comments

F Gambusia affinis 96-h survival 240 N/A Wallen & Lasater 1957 (Taken from US EPA Ecotox databases)

C Daphnia magna 72-h survival 102 N/A Dowden & Bennett (1965) (Taken from US EPA Ecotox databases)

C Daphnia magna 96-h survival 102 N/A Freeman L. and Fowler I. (1953) (Taken from US EPA Ecotox databases)

C Daphnia magna 24-h survival 171 N/A Dowden B.F. and H. Bennett 1965 (Taken from US EPA Ecotox databases)

F= Fish, C = Crustacean, A = Algae

Table 3 summarizes acute aquatic toxicity tests performed by Nautilus Environmental (Nautilus, 2009) on the BalPure ® system at differing hypochlorite concentrations. These tests were done in conjunction with studi es performed at the University of Washington and were not intended for evaluation of efficac y. Rather, the tests were to evaluate aquatic toxicity and disinfection by-products, and the applied hypochlorite c oncentrations were based on the organic matter present in the test water. Tests were conducted using algae, inverteb rates, and fish in fresh water, brackish water, and saltwater. Al l tests followed OECD or US EPA OPPTS testing protocols. The samples were collected for toxic ity testing on Day 0 (imme diately after addition of hypochlorite), and on Day 2 and Day 5 after treatment. Following th e collection of Day 5 samples, residual chlorine was neutralized with sodium bisulfite and a second Day 5 post-neutralization sample was collected for toxicity testing. The neutralized water was then held an additional 5 days (i.e. 10 days after hypochlor ite treatment) and then evaluated for aquatic toxicity. With the exception of the marine algae, Skeletonema, results indicated that the neutralization step removed toxi city. The water quality data associated with the samples collected for toxicity testing and the testing resu lts are discussed further in section 11 “Evaluation of Treated Ballast Water”.
Documentation of the quality assurance program fo r toxicity testing at Nautilus Environmental is included in appendix A.7. 1

1 Appendix A.7 is provided in the confidential dossier of the proposal for approval.MEPC 60/2/9 ANNEX Page 22

I:\MEPC\60\2-9.doc Table 3 : Acute Aquatic Toxicity-Treated Ballast Water

Test Substance F C A Species Days after treatment,

Pre/Post neutralization Test Type E(L)C50 % sample NOEC % sample References/ comments

Freshwater (average conductivity 1002 μS/cm)

0 72h growth (OECD

201) 7.76 <6.25 Nautilus Environmental, 2009

2 72h growth (OECD

201) 11.5 <6.25 Nautilus Environmental, 2009

5-pre 72h growth (OECD

201) >100 25 Nautilus Environmental, 2009

5-post* 72h growth (OECD

201) >100 100 Nautilus Environmental, 2009 A Selenastrum capriconutum

5+* 72h growth (OECD

201) >100 100 Nautilus Environmental, 2009

0 48h survival (OECD

202) 3.13 <6.25 Nautilus Environmental, 2009

2 48h survival (OECD

202) 3.13 <6.25 Nautilus Environmental, 2009

5-pre 48h survival (OECD

202) 41.4 25 Nautilus Environmental, 2009

5-post* 48h survival (OECD

202) >100 100 Nautilus Environmental, 2009 6.7 mg/L HOCl

C Daphnia
magna

5+* 48h survival (OECD

202) >100 100 Nautilus Environmental, 2009MEPC 60/2/9 ANNEX Page 23

I:\MEPC\60\2-9.doc Test Substance F C A Species Days after treatment,

Pre/Post neutralization Test Type E(L)C50 % sample NOEC % sample References/ comments

0 96h survival (OECD

203) 4.03 <6.25 Nautilus Environmental, 2009

2 96h survival (OECD

203) 43.8 25 Nautilus Environmental, 2009

5-pre 96h survival (OECD

203) >100 100 Nautilus Environmental, 2009

5-post* 96h survival (OECD

203) >100 100 Nautilus Environmental, 2009 F Pimephalus promelas

5+* 96h survival (OECD

203) >100 100 Nautilus Environmental, 2009

Brackish water (average salinity 22.14 PSU)

0 96h survival

(EPA OPPTS 850.5400) 8.61 <6.25 Nautilus Environmental, 2009

2 96h survival

(EPA OPPTS 850.5400) 15.9 12.5 Nautilus Environmental, 2009

5-pre 96h survival

(EPA OPPTS 850.5400) 16.4 12.5 Nautilus Environmental, 2009

5-post* 96h survival

(EPA OPPTS 850.5400) 66.1 50 Nautilus Environmental, 2009 9.1 mg/L HOCl

A Skeletonema

costatum

5+* 96h survival

(EPA OPPTS 850.5400) >100 100 Nautilus Environmental, 2009MEPC 60/2/9 ANNEX Page 24

I:\MEPC\60\2-9.doc Test Substance F C A Species Days after treatment,

Pre/Post neutralization Test Type E(L)C50 % sample NOEC % sample References/ comments

0 96h survival

(EPA OPPTS 850.1035) 5.94 6.25 Nautilus Environmental, 2009

2 96h survival

(EPA OPPTS 850.1035) 17.7 12.5 Nautilus Environmental, 2009

5-pre 96h survival

(EPA OPPTS 850.1035) 17.5 12.5 Nautilus Environmental, 2009

5-post* 96h survival

(EPA OPPTS 850.1035) >100 100 Nautilus Environmental, 2009 C Americamysis bahia

5+* 96h survival

(EPA OPPTS 850.1035) >100 100 Nautilus Environmental, 2009

0 96h survival

(EPA OPPTS 850.1075) 11.7 6.25 Nautilus Environmental, 2009

2 96h survival

(EPA OPPTS 850.1075) 28.4 25 Nautilus Environmental, 2009

5-pre 96h survival

(EPA OPPTS 850.1075) 22.6 12.5 Nautilus Environmental, 2009 F Cyprinodon variegatus

5-post* 96h survival

(EPA OPPTS 850.1075) >100 100 Nautilus Environmental, 2009MEPC 60/2/9 ANNEX Page 25

I:\MEPC\60\2-9.doc Test Substance F C A Species Days after treatment,

Pre/Post neutralization Test Type E(L)C50 % sample NOEC % sample References/ comments

5+* 96h survival

(EPA OPPTS 850.1075) >100 100 Nautilus Environmental, 2009

Saltwater (average salinity 31 PSU)

0 96h survival

(EPA OPPTS 850.5400) 33.4 25 Nautilus Environmental, 2009

2 96h survival

(EPA OPPTS 850.5400) 34.5 25 Nautilus Environmental, 2009

5-pre 96h survival

(EPA OPPTS 850.5400) 38 25 Nautilus Environmental, 2009

5-post* 96h survival

(EPA OPPTS 850.5400) >100 100 Nautilus Environmental, 2009 A Skeletonema

costatum

5+* 96h survival

(EPA OPPTS 850.5400) >100 100 Nautilus Environmental, 2009

0 96h survival

(EPA OPPTS 850.1075) 10.5 6.25 Nautilus Environmental, 2009

2 96h survival

(EPA OPPTS 850.1075) 16.7 12.5 Nautilus Environmental, 2009 4.3 mg/L HOCl

C Americamysis bahia

5-pre 96h survival

(EPA OPPTS 850.1075) 16.8 12.5 Nautilus Environmental, 2009MEPC 60/2/9 ANNEX Page 26

I:\MEPC\60\2-9.doc Test Substance F C A Species Days after treatment,

Pre/Post neutralization Test Type E(L)C50 % sample NOEC % sample References/ comments

5-post* 96h survival

(EPA OPPTS 850.1075) >100 100 Nautilus Environmental, 2009

5+* 96h survival

(EPA OPPTS 850.1075) >100 100 Nautilus Environmental, 2009

0 96h survival

(EPA OPPTS 850.1075) 24.3 25 Nautilus Environmental, 2009

2.5-pre 96h survival

(EPA OPPTS 850.1075) 28.3 12.5 Nautilus Environmental, 2009

5-pre 96h survival

(EPA OPPTS 850.1075) 35 25 Nautilus Environmental, 2009

5-post* 96h survival

(EPA OPPTS 850.1075) >100 100 Nautilus Environmental, 2009 4.3 mg/L HOCl

F Cyprinodon variegatus

5+* 96h survival

(EPA OPPTS 850.1075) >100 100 Nautilus Environmental, 2009

F= Fish, C = Crustacean, A = Algae; *Samples neutralized with sodium bisulfite.MEPC 60/2/9 ANNEX Page 27

I:\MEPC\60\2-9.doc 3.2 Chronic aquatic toxicity

The following table summarizes data found in litera ture concerning chronic aquatic toxicity for the substances associated with the BalPure ® system. The available chronic data was limited for some chemicals. Chronic toxicity data for treated ballast water is presente d in Table 5 below.

Table 4 : Overview of Chronic Aquatic Toxicity

Test Substance F

C

A Species Test Type EC50 mg/L NOEC mg/L Reference/ comments

Active Substance

F Ictalurus punctatus 134-d growth N/A 0.005 Hermanutz et al. (1990)

C Gammarus pseudolimnaeus 70-d fecundity N/A 0.012 Arthur et al. (1975) TRO (Freshwater)

A Natural algae 24-d inhibition of biomass N/A 0.079 Pratt et al (1988)

F Menidia peninsula 32-d egg to posthatch N/A 0.04 Goodman et al. (1983)

C Panaeus kerathurus 7-d growth/molting N/A 0.079 Saroglia and Scarano (1979) TRO (Saltwater)

A Natural phytoplankton 21-d reduction in cell density N/A 0.001 Sanders et al. (1981)

F Oncorhynchus kisutch 23-d survival N/A 0.101 Holland et al. (1960) Hypochlorous acid C Americamysis bahia 7-d reproduction N/A 0.048 Fisher et al. (1994) Hypobromous acid No data found in literature Relevant Chemicals

F Morone saxatilis 10-d survival 93 50 Richardson et al. (1981)

Bromate A Isochrysis galbana 7-d growth inhibition N/A 8 Erickson & Freeman (1978) (taken from US EPA Ecotox databases) THMs

F Oryzias latipes 9- month growth N/A 1.5 Toussaint et al. (2001)

C Daphnia
magna 21-d reproduction N/A 13 Kuhn et. al (1988) (Taken from US EPA Ecotox databases) Chloroform

A Skeletonema

costatum 5-d reduction in cell volume 477 216 Cowgill et al. (1989)MEPC 60/2/9 ANNEX Page 28

I:\MEPC\60\2-9.doc Test Substance F

C

A Species Test Type EC50 mg/L NOEC mg/L Reference/ comments

F Cyprinodon variegates 28d- growth N/A 8.5 Ward et al. 1981(Taken from US EPA from Ecotox databases) Bromoform

A Pseudokirchne riella subcapitata 96-h biochemistry change N/A 26.1 U.S. EPA (1978) (Taken from US EPA Ecotox databases)Dichlorobromomethane No data found in literatureChlorodibromomethane No data found in literature HAAs

Dichloroacetic acid A Scenedesmus subspicatus 7-d cell proliferation 1485 (EC0

3) N/A Trenel and Kuhn (1982) (from ECOTOX databases) Dibromoacetic acid No data found in literature

A Scenedesmus quadricauda 7-d growth threshold N/A 200 OECD SIDS (2008) Trichloroacetic acid A Chlorella vulgaris 4d-population change N/A 100 Garten (1990) (taken from US EPA Ecotox databases)Bromochloroacetic acid No data found in literature

C Daphnia
magna 21d-NOEC N/A 3.2 Kuhn (1988) (Taken from US EPA Ecotox databases)Monobromoacetic acid

A Scenedesmus subspicatus 3d-population change N/A 1.4 Kuhn & Pattard (1990) (Taken from US EPA Ecotox databases)

Monochloroacetic acid C Daphnia
magna 21d-reproduction change N/A 32 Kuhn & Pattard (1990) (Taken from US EPA Ecotox databases)MEPC 60/2/9 ANNEX Page 29

I:\MEPC\60\2-9.doc Test Substance F

C

A Species Test Type EC50 mg/L NOEC mg/L Reference/ comments

A Scenedesmus subspicatus 7d-histology (EC0 3) 0.13 N/A Trenel and Kuhn (1982) (Taken from US EPA Ecotox databases)

Other Chemicals Sodium bisulfite No data found in literature F= Fish, C = Crustacean, A = Algae

Chronic toxicity tests were conducted by Nautilu s Environmental (Nautilus, 2007) using treated ballast water samples collected during the full-s cale study at the Naval Research Facility in Key West, Florida (United States). The applied hypochlorite concentr ation was based on the level of organic matter present in the test water. Samp les were collected from challenge tests in which organism and organic enriched ballast water wa s treated with hypochlorite to a 15 mg/L total residual oxidant (TRO) concentrat ion and then held for 48 hours (2 days). After 48 hours the treated water was neutralized with an excess con centration of ~ 2 mg/L s odium bisulfite. Five concentrations of treated sample using a 0.5X dilution series were tested. Bioassays were conducted using 7-day tests with a mysid ( Americamysis bahia ) and a larval fish ( Menidia beryllina ), a 48-hour larval development test using a mussel ( Mytilus galloprovincialis ), and a 48-hour germination and germ tube length test using the giant kelp ( Macrocystis pyrifera ). Chronic toxicity tests using mysid shrimp a nd silverside minnows we re conducted according to US EPA (2002) methods. Chronic toxicity te sts using mussels and giant kelp were conducted according to US EPA (1995) methods. Document ation of the quality assurance program for toxicity testing at Nautilus Enviro nmental is included in appendix A.7 2

. Results for the chronic toxicity tests are summar ized in Table 5 below a nd discussed further in section 11 “Evaluation of Treated Ballast Water” . No toxicity was observed with the treated water samples for any species tested. The highest concentration with no observed effect (NOEC) was 100 percent sample (Nautilus, 2007).

2 Appendix A.7 is provided in confidential dosier of the proposal for approval.MEPC 60/2/9 ANNEX Page 30

I:\MEPC\60\2-9.doc Table 5 : Chronic Aquatic Toxicity-Treated Ballast Water

Test Substance F C A Species Days after hypochlorite treatment, plus neutralization Test Type E(L)C50 % Sample NOEC % Sample References/ comments

Seawater (average salinity 36 PSU)

F Menidia beryllina 2 days 7 day survival(EPA R821-02- 014) >100 100 Nautilus Environmental, 2007

F Menidia beryllina 2 days 7 day survival(EPA R821-02- 014) >100 100 Nautilus Environmental, 2007

C Americamysis bahia 2 days 7 day survival(EPA R821-02- 014) >100 100 Nautilus Environmental, 2007

C Americamysis bahia 2 days 7 day survival(EPA R821-02- 014) >100 100 Nautilus Environmental, 2007

B Mytilus galloprovincialis 2 days 48 hour larval developme nt (EPA 600-R-95- 136) >100 100 Nautilus Environmental, 2007

A Macrocystis pyrifera 2 days 48 hour germination (EPA 600- R-95-136) >100 100 Nautilus Environmental, 2007 15.0 mg/L HOCl

A Macrocystis pyrifera 2 days 48 hour germ tube length (EPA 600- R-95-136) >100 100 NautilusEnvironmental, 2007

F = Fish, C = Crustacean, B = Bivalve, A = AlgaeMEPC 60/2/9 ANNEX Page 31

I:\MEPC\60\2-9.doc 3.3 Endocrine disruption

Endocrine disrupting chemicals (EDCs) are define d as “exogenous substances that cause adverse effects in an organism or its progeny, consequent to changes in endocrine functions”. In a recently conducted study to investigate whether chlorinated by-products formed through wastewater disinfection were estrogenic, no relationship wa s found between the formation of THMs and estrogenic activity (Schiliro 2009). In fact, chlorination of surface water and effluents was found to decrease estrogenic activity, which is thought to be due to the oxidation effects of chlorine (Lee, 2004). A thorough review of the litera ture found no indications that either of the Active Substances in the TRO (HOCl/HOBr) or the chemically more complex THMs/HAAs were EDCs.

3.4 Sediment toxicity

The sediment toxicity of a chemical is a function of the ability of the chemical to be adsorbed to sediment as well as its persiste nce and toxicity. The organic car bon partition coefficient (Koc) is a measure of the tendency of an organic subs tance to be adsorbed by soil or sediment. The TRO Active Substances are all inorganic substances and therefore have a very low calculated Koc as presented in Table 6 below. None of the organic Relevant Chemicals (THMs/HAAs) appear to have the pot ential to be absorbed to se diments to a significant extent based on their low Kocs (<500 L/kg) and other in formation in the lite rature (HSDB/TOXNET). The low Kocs for all of the chemicals in Table 6 indicate that the chemicals would not readily partition to sediment and therefore be of concer n to benthic organisms residing in the sediment.

Table 6 : K OC s for Active Substances and Relevant Chemicals

Chemical Koc(L/kg) Reference Active Substances TRO NA
Hypochlorous acid 13.2 As calculated

− EPI Suite (2009c)
Hypobromous acid 13.2 As calculated

− EPI Suite (2009b) Relevant Chemicals

Bromate 31.8 As calculated

− EPI Suite (2009a) THMs Chloroform 153-196 HSDB/TOXNET (2009f) Bromoform 35 HSDB/TOXNET (2009l) Dichlorobromomethane 9 HSDB/TOXNET (2009c) Chlorodibromomethane 84 HSDB/TOXNET (2009e) HAAs Bromochloroacetic acid 1.9 HSDB/TOXNET (2009b)

Monochloroacetic acid 31 HSDB/TOXNET (2009d) Monobromoacetic acid 1.9 HSDB/TOXNET (2009a) Dichloroacetic acid 75 HSDB/TOXNET (2009h)

Trichloroacetic acid 130 HSDB/TOXNET (2009m)MEPC 60/2/9 ANNEX Page 32

I:\MEPC\60\2-9.doc Chemical Koc(L/kg) Reference Dibromoacetic acid 1.5 HSDB/TOXNET (2009g) Other Chemicals

Sodium bisulfite 2.2 As calculated

− EPI Suite (2009d)

3.5 Bioavailability/biomagnification/bioconcentration

The bioconcentration factor (BCF) is the concentr ation of a particular chemical in a tissue per concentration of chemical in wa ter (reported as L/kg) . This physical prope rty characterizes the accumulation of pollutants through chemical parti tioning from the aqueous phase into an organic phase, such as fish or other aquatic organism tissues. The BCF is the best measure of bioavailability, biomagnification, and bioconcentration. Typical BCFs for organic chemicals in fish and most aquatic invertebrates are in the 500 – 1000 range. The BCFs summarized in Table 7 below are all much lower i ndicating a low potential for significant bioconcentration/bioaccumulation in a quatic organisms (HSDB/TOXNET). Little biomagnification should occur in a food web since the chemicals ar e not readily bi oconcentrated or bioaccumulated.

Table 7 : BCFs for Active Substances and Relevant Chemicals

Chemical BCF (L/kg) Reference A ctive Substances TRO NA H yp ochlorous aci d 3.2 As calculated

− EPI S uite H yp obromous aci d 3.2 As calculated

− EPI S uite R elevant Chemicals Bromate 3.2 As calculated

− EPI Suite THMs Chloroform 2.9-10.35 HSDB/TOXNET ( 2009f ) Bromoform 14 HSDB/TOXNET ( 2009l ) Dichlorobromomethane 7 HSDB/TOXNET ( 2009c ) Chlorodibromomethane 9 HSDB/TOXNET ( 2009e ) HAAs Bromochloroacetic aci d 3.2 HSDB/TOXNET ( 2009b ) Monochloroacetic aci d 3.1 HSDB/TOXNET ( 2009d ) Monobromoacetic aci d 3.2 HSDB/TOXNET ( 2009a ) Dichloroacetic aci d 0.3 HSDB/TOXNET ( 2009h ) Dibromoacetic aci d 0.17 HSDB/TOXNET ( 2009 g) Trichloroacetic aci d 0.1-1.7 HSDB/TOXNET ( 2009m )

Other Chemicals Sodium Bisulfite 3.2 As calculated

− EPI SuiteMEPC 60/2/9 ANNEX Page 33

I:\MEPC\60\2-9.doc 3.6 Food web/population effects

Population and food web effects can occur if substances have properties that tend to bioaccumulate and/or persist in the environmen t. Little biomagnification and persistence in aquatic and mammalian food webs is an ticipated from the Active Substances or Relevant Chemicals. This conclusion is base d on the low Kocs of the chemicals. All of the Kocs for the Active Substances and Relevant Chemicals (section 3.4) are <500 L/kg which indicate that the chemicals woul d not readily absorb to sediment and therefore be a concern to benthic populations and their food webs. Si milarly, the BCFs also show that little bioconcentration/bioaccumulation will occur in aquatic organisms (section 3.5).

4 DATA ON MAMMALIAN TOXICITY (G9: 4.2.1.2)

The Tables 8 through 14 below present data on mammalian toxicity for different endpoints and effects. The information was located through litera ture reviews of existing data from a variety of governmental organizations such as the World H ealth Organization, the International Programme on Chemical Safety, and the United States Envi ronmental Protection Agency. In some cases, data was gathered from Material Safety Data Sheets (MSDS).

In the studies reviewed, the chemicals were tested for various exposure routes (oral, dermal, etc.) and/or chemical forms, which do not necessarily reflect the actual poten tial exposure routes or chemical forms associated with the BalPure ® system. Potential for human exposure to the substances associated with the BalPure ® system is discussed in detail in section 12.2.

4.1 Acute toxicity

Table 8 : Acute Mammalian Toxicity Data

Test Substance Exposure Route Species Value range Reference/ Comments Active Substance Hypochlorous acid (test concentration not specified) a. Oral LD50 b. Oral LD50 a. Rat b. Mouse a. 8900 mg/kg b. 5800 mg/kg HSDB/TOXNET (2009i)

Hypobromous acid No data found in literature. Thought to have similar toxicity to chlorinated analogue. Relevant Chemicals

Bromate Oral LD50 Rats, mice and hamsters 280-495 mg/kg IPCS (2000a)

HAAs

Bromochloro-acetic acid 2-Week Oral Drinking Water Study a. Rat b. Mice a. NOEL = 140 mg/kg/day b. NOEL = 170 mg/kg/day NTP (2009)MEPC 60/2/9 ANNEX Page 34

I:\MEPC\60\2-9.doc Test Substance Exposure Route Species Value range Reference/ Comments

Monochloro-acetic acid a. Oral LD50 b. Skin LD50 c. Oral LD50 a. Mouse b. Rabbit c. Mice, male rats, and male guinea pigs a. 260 mg/kg b. 230 mg/kg c. 255, 76, and 80 mg/kg respectively a,b. IPCS (2000a) c. WHO (2004b)

Monobromo-acetic acid Oral LD50 a. Rat 177 mg/kg ICPS (2000a)

Dichloroacetic acid a. Oral LD50 b. Oral LD50 a. Rat b. Mouse a. 4500 mg/kg b. 5500 mg/kg IPCS (2000a)

Dibromoacetic Acid Oral LD50 a. Rat 1737 mg/kg WHO (2004a)

Trichloroacetic acid a. Oral LD50 b. Oral LD50 a. Rat b. Mouse a. 3300 mg/kg b. 4900 mg/kg IPCS (2000a)

THMs

Chloroform a. Oral LD50 b. Oral LD50

c. Inhalation LC50, 6 h
exposure a. Rat b. Mice c. Rats a. 450-2000 mg/kg b. 36-1366 mg/kg c. 9.2 g/m 3 IPCS (2004)

Bromoform a. Oral LD50 b. Oral LD50 a. Male rats/female rats b. Male mice/female mice a. 1388 mg/kg/1147 mg/kg b. 1400 mg/kg/1550 mg/kg IPCS (2000a)

Dichlorobromomethane Oral LD50 Male rats/female rats 430 mg/kg/510 mg/kg ATSDR & US EPA (1989)

Chlorodibromomethane a. Oral LD50 b. Oral LD50 a. Male Rats/female rats b. Male mice/female mice a. 1186 mg/kg/848 mg/kg b.800 mg/kg/1200mg/kg IPCS (2000a)

Other Chemicals

Sodium bisulfite Oral LD50 Rat 2000 mg/kg Health Council of the Netherlands (2005)MEPC 60/2/9 ANNEX Page 35

I:\MEPC\60\2-9.doc 4.2 Effects on skin and eye

Table 9 : Skin, Eye and Dermal Effects

Test Substance Skin Eye Sensitization Reference/ comments Active Substance

Hypochlorous acid Irritant at concentrations ≥ 5% Irritant at concentrations ≥ 5%; concentrations ≥ 12.5% corrosive to
eye Low potential for sensitization after chronic exposure to damaged skin; in Patch tests on humans show positive results for concentrations ranging from 0.3-2% HSDB/TOXNET (2009i), EU (2007b)

Hypobromous acid No data found in literature. Thought to have similar toxicity to chlorinated analogue. Relevant Chemicals Bromate (as sodium bromate) Causes irritation Causes irritation No data found in literature HSDB/TOXNET (2009k)

THMs

Chloroform Causes irritation (redness and pain) (10μg/m 3 ) Causes irritation Possible damage to nervous system, heart, liver, and kidneys; Possible dermatitis IPCS (2004)

Bromoform Causes irritation Causes irritation Greater risk sensitization forthose with preexisting skin/respiratory problems U.S. Dept. Health and Human Services & ATSDR (2005)

Dichlorobromomethane Causes irritation; Levels of 13-34 μg/L show a measurable body burden for humans (from chlorinated swimming pools) Causes irritation No data found in literature HSDB/TOXNET (2009c), NTP 1991MEPC 60/2/9 ANNEX Page 36

I:\MEPC\60\2-9.doc Test Substance Skin Eye Sensitization Reference/ comments

Chlorodibromomethane Causes irritation Causes irritation No data found in literature U.S. Dept. Health and Human Services & ATSDR (2005) HAAsBromochloroacetic acid Causes skin burns Causes eye burns No data found in literature HSDB/TOXNET (2009b)

Monochloroacetic acid Corrosive, causes skin burns; concentrations above 40% are toxic to dermal tissue Corrosive,
causes eye burns and blurred vision; 0.9% solution

causes severe eye irritation Liver and kidney damage after chronic
exposure Mallinckrodt Baker MSDS (2006a), EU (2005)

Monobromoacetic acid Causes skin burns Causes eye burns Liver, kidney, and muscle damage after chronic exposure HSDB/TOXNET (2009a)

Dichloroacetic acid Redness, blisters, serious skin burns Redness, severe burns No data found in literature IPCS (2000c)

Dibromoacetic acid Causes skin burns Causes eye burns No data found in literature Sigma Aldrich MSDS (2009)

Trichloroacetic acid Corrosive,

causes irritation, and moderate burns Corrosive,

causes irritation and moderate burns No data found in literature ATSDR & OSHA (1989)

Other Chemicals

Sodium bisulfite NOAEL 0.5% solution Causes irritation No data found in literature Mallinckrodt Baker MSDS (2006b), EU (2000)

Note : Test concentrations are provided when specified in the reference document.MEPC 60/2/9 ANNEX Page 37

I:\MEPC\60\2-9.doc 4.3 Repeated-dose toxicity

Table 10 : Repeated-dose toxicity

Chemical Exposure Route Species Value Range Reference/ Comments Active Substances

Hypochlorous acid a. Oral, 90-day, drinking water b. Oral, 90-day, drinking water a. Rats b. Mice a. NOAEL, 10 mg/kg-day b. Spleen/liver weights depressed in males, but not females at the higher doses* IPCS (2000a)

Hypobromous acid No data found in literature. Thought to have similar toxicity to chlorinated analogue. Relevant Chemicals

Bromate Oral, drinking water, 13 weeks, doses 0-10,00- mg/L Rats b. LOAEL=63 mg/kg bw/d (body weight gain, serum parameters, kidney effects) IPCS (2000a) US EPA (2001a)

THMs

Chloroform a. Oral, gavage, 90 d, 0-270 mg/kg/d b. Oral, drinking water, 1-6 months, 0-1,800 mg/L (0-263 mg/kg) Mice a. LOAEL 60 mg/kg (liver effects) b. NOAEL 34 mg/kg a. IPCS (2000a) b. US EPA (2001b)

Bromoform Gavage, 13 weeks Rats and mice NOAEL, 25 mg/kg/day (rats) & 100 mg/kg/day (mice) IPCS (2000a)

Dichlorobromomethane Oral, gavage, 13 weeks, a. 0-300 mg/kg/d b. 0-400 mg/kg/d a. Rats b. Mice a. NOAEL 75 mg/kg/d (decrease in body weight) b. NOAEL 50 mg/kg/d (kidney lesions) NTP (1987)

Chlorodibromomethane a. gavage, 13 weeks b. gavage, 90 days a. Rats and mice b. Rats a. NOAEL 30 mg/kg/day b. kidney damage at 50 & 100mg/kg/day* IPCS (2000a)

HAAs

Bromochloroacetic acid Drinking water, 3 months, 0-1000 mg/L a. 0-85 mg/kg b. 0-140 mg/kg a. Rats b. Mice a. NOAEL 20 mg/kg/d (increased liver weight) b. LOAEL 8 mg/kg/d (hematopoietic cell proliferation in the spleen) NTP (2009)MEPC 60/2/9 ANNEX Page 38

I:\MEPC\60\2-9.doc Chemical Exposure Route Species Value Range Reference/ Comments

Monochloro -acetic acid Oral gavage, 13 weeks a. 0-150 mg/kg/d b. 0-200 mg/kg/d a. Rats b. Mice a. LOAEL 30 mg/kg/d (changes in heart, liver, and kidney weights, clinical chemistry values) b. NOAEL 100/LOAEL 150 mg/kg/s (increased liver weight) NTP (1992b), EU (2005)

Monobromoacetic acid No data available

Dichloroacetic acid a. Oral, 90 days, doses up to 72 mg/kg/d b. oral, 30 days c. oral, 90 days a. Dogs b. Mice c. Rats a. LOAEL 12.5 mg/kg/d (liver effects, testicular degeneration) b. LOAEL 70-100 mg/kg/day c. LOAEL 125 mg/kg/day IPCS (2000a)

Dibromoacetic acid Oral, drinking water, 3 months, 0-2000 mg/L a. Rats b. Mice Increased liver weights a. LOAEL 10 mg/kg/d b. LOAEL 56 mg/kg/d, NOAEL 30 mg/kg/d NTP 2007

Trichlor acetic acid a. Oral, 90 days, 0-355 mg/kg/day b. Oral, 90 days, 785 mg/kg/day a. Rats b. Rats a. decrease in body weight, NOAEL 36.5 mg/kg/d b. minimal evidences of liver damage* IPCS (2000a)

Other Chemicals

Sodium bisulfite Oral, drinking water, 5-8 week study Rats NOAEL 70 mg/kg bw/day Health Council of the Netherlands (2005)
*No NOAEL/LOAEL reportedMEPC 60/2/9 ANNEX Page 39

I:\MEPC\60\2-9.doc 4.4 Chronic toxicity

The data located in a literature search for chronic mammalian studies (six months or longer) is presented below.

Table 11 : Chronic Mammalian Toxicity

Test Substance Exposure Route Species Value range Reference/ comments Active Substance Hypochlorous acid Oral, drinking water, 2-year Mice and
rats NOAEL= 14 mg/kg/d Dilation of renal tubules in male mice IPCS (2000a) & NTP (1992a)

Hypobromous acid No data found in literature. Thought to have similar toxicity to chlorinated analogue. Relevant Chemicals

Bromate Oral, drinking water, 100 weeks a. 0-400 mg/L b. 0-800 mg/L a. Rats b. Mice a. kidney effects, NOAEL 1.1 mg/kg/d b. no effects, NOAEL >60 mg/kg/d US EPA (2001a)

THMs

Chloroform a. Oral, gavage, 2-years b. Oral, 7.5 years, toothpaste a. Mice and
rats b. Dogs a. liver effects noted, including tumors* b. LOAEL, 15 mg/kg/day (6 days/week) a. IPCS (2004) b. US Dept. of Health and Human Services in ATSDR (1997)

Bromoform Gavage, 2-year, doses up to 200 mg/kg/day Mice and
rats LOAEL 100 mg/kg/d (liver effects) IPCS (2000a) US Dept. Health and ATSDR (2005)

Dichlorobromomethane a. Gavage, 104 week study, doses 5 days/week b. Gavage, 104 week study, doses 5 days/week a. Rats b. Mice a. NOAEL 100 mg/kg/day b. LOAEL 25 mg/kg/day (renal cytomegaly) a. ATSDR & US EPA (1989) b. US EPA (1991c)

Chlorodibromomethane Oral gavage, 2-years a. 0/40/80 mg/kg/d b. 0/50/100 mg/kg/d a. Rats b. Mice Kidney and liver effects noted at all doses* IPCS (2000a)MEPC 60/2/9 ANNEX Page 40

I:\MEPC\60\2-9.doc Test Substance Exposure Route Species Value range Reference/ comments HAAs

Bromochloroacetic acid Oral, drinking water, 2 years, 0-1000 mg/L a. 0-50 mg/kg b. 0-90 mg/kg a. Rats b. Mice a. NOAEL 13 mg/kg/d (non-neoplastic lesions in the liver) b. LOAEL 15 mg/kg/d (non-neoplastic and neoplastic liver effects) NTP (2009)

Monochloroacetic acid Oral, drinking water, 104 weeks, 0-60 mg/kg/d Rats NOAEL 3.5/LOAEL 26 mg/kg/d (changes in body weight) EU (2005)

Monobromoacetic acid No long-term toxicity studies for any exposure route were identified WHO (2004a)

Dichloroacetic acid Oral, drinking water, 2 years, doses up to 2 g/L a. Rats b. Mice a. Dose dependent increase in liver tumor incidence, LOAEL 40 mg/kg/d, NOAEL 4 mg/kg/d b. 80% increase in liver tumor incidence at 95 mg/kg/d IPCS (2000a)

Dibromoacetic acid Oral, drinking water, 2 years, 0-1000mg/L a. Rats b. Mice a. LOAEL 2mg/kg/d (liver effects in males, kidney effects in females) b. liver tumors, male: LOAEL 4/mg/kg/d, female: LOAEL 35/NOAEL 4 mg/kg/d NTP (2007)

Trichloroacetic acid Oral, 2-year, doses up to 5g/L a. Rats b. Mice a. liver effects, NOAEL 32.5 mg/kg/d b. dose-dependent increase in liver tumors, NOAEL 40 mg/kg/d IPCS (2000a) Health Canada (2009)

Other Chemicals

Sodium bisulfite a. Oral, 3 generation study, doses from 0.125-20 g/L a. Oral, doses from 0.1%-2% conc. for 2 years a. Rats b. Rats a. No evidence of systemic toxicity at levels of 10 g/L or 20 g/L; calculated NOAEL 72/kg/day b. LOAEL 0.1%. Reduced survival and body weight, myocardial fibrosis, and renal calcification at doses of 0.25% and higher a. Health Council of the Netherlands (2005) b. HSDB/TOXNET (2009j)

*No NOEAEL/LOAEL reportedMEPC 60/2/9 ANNEX Page 41

I:\MEPC\60\2-9.doc 4.5 Developmental and reproductive toxicity

Table 12 : Summary of Developmental and Reproductive Toxicity Data

Chemical Exposure Route Species Effects Reference/ Comments Active substances

Hypochlorous acid Oral, chlorinated drinking water, 10 mg/kg/day for 7 generations Rats No effects on fertility, growth, or survival IPCS (2000a)

Hypobromous acid No data found in literature. Thought to have similar toxicity to chlorinated analogue. Relevant chemicals

Bromate (as sodium bromate) Oral, drinking water, 35d, doses up to 250 mg/L Rats Male fertility: decreased sperm density, NOAEL 7.7 mg/kg/d, female fertility: no effects, NOAEL >22 mg/kg/d US EPA (2001a)

THMs

Chloroform Doses up to 41 mg/kg/day Mice No effect on fertility or fetal development IPCS (2004)

Bromoform Gavage, doses ranging from 0-200 mg/kg/day Mice postnatal survival significantly decreased at doses of 200 mg/kg/day. No other reproductive effects seen in F1 or F2 generations* US EPA (1991b)

Dichlorobromomethane Oral, 10 day study, doses from 50-200 mg/kg/d ay on days 6-15 of gestation Rats Doses of 50 mg/kg/day show fetotoxic effects* ATSDR and US EPA (1989)

Chlorodibromomethane Oral, 2- generation study, doses up
to 685 mg/kg/day Mice Decrease in fertility in high dose groups. No lethal or teratogenic effects* IPCS (2000a)

HAAs

Bromochloroacetic acid Oral, gavage, 14 days, 0/8/24/72 mg/kg/d Rats Decreased sperm fertility at all doses Klinefelter et.al. (2002)MEPC 60/2/9 ANNEX Page 42

I:\MEPC\60\2-9.doc Chemical Exposure Route Species Effects Reference/ Comments

Monochloroacetic acid Gavage, doses of 0 or 193 mg/kg/day on gestation days 1-22 Rats Significant decrease in body weight but no adverse reproductive, developmental, or teratogenic effects reported. LOAEL of 193 mg/kg/day for maternal toxicity WHO (2004b)

Monobromoacetic acid Oral intubation, doses ranging from 0-100mg/kg/day on gestation days 6-15 Rats Maternal weight changes, decrease in fetal weight, LOAEL of 100 mg/kg/day HSDB/TOXNET (2009a)

Dichloroacetic acid a. Gavage, 10 weeks, doses up to 125 mg/kg/day b. Gavage, days 6-15 of gestation a. Rats b. Rats a. dose-dependent effect on male reproductive function, LOAEL 31 mg/kg/d (reduced weights of accessory organs) b. soft tissue abnormalities in fetal rats, LOAEL 140 mg/kg/d, NOAEL 14 mg/kg/d IPCS (2000a)

Dibromoacetic acid Gavage, doses up to 250 mg/kg/day Rats Effects noted on male reproductive behavior

> 10 mg/kg/d), sperm morphology (> 50 mg/kg/d), fertility (> 250 mg/kg/d) IPCS (2000a)

Trichloroacetic acid Gavage, doses up to 1800 mg/kg/day, gestation day 6-15 Rats Reduced pup weights ≥ 800 mg/kg/d, soft tissue malformations ≥ 330 mg/kg/d (LOAEL) IPCS (2000a)

Other Chemicals

Sodium bisulfite a. Topical treatment 2 mL/kg on gestational days 1-19 b. Gavage, doses of 100- 120 mg/kg bw during organogenesis a. Female
rats b. Rats, mice, rabbits, hamsters a. mean number of corpora lutea, implantation sites, live fetuses, and resorptions per pregnancy and number of litters w/ resorptions not significantly affected* b. Did not induce maternal/reproduction toxicity* Health Council for the Netherlands (2005)

*No NOEAL/LOAEL reportedMEPC 60/2/9 ANNEX Page 43

I:\MEPC\60\2-9.doc 4.6 Carcinogenicity
Table 13 below summarizes the carcinogenic cl assification for the chemicals related to the BalPure ® system. The majority of the classifications are mandated by the International Agency for Research on Cancer (IARC) and the US EPA, while some come fro m other governmental organizations such as Health Canada. Some ch emicals have not been cl assified by the IARC or other governmental organizations due to a lack of sufficient research an d data. Most of the chemicals that are classified are characterized as possibly or unlikely to be carcinogenic to humans.
Table 13 : Summary of Data on Carcinogenicity

Chemical Carcinogen Classification Description Reference/ Comments Active substances

Hypochlorous acid a. IARC group 3-not classifiable as carcinogenic to humans

b. Not classified by NTP a. Inadequate evidence in experimental animals
b. Oral exposure in drinking water, 2 years, at doses a) 0- 14.4 mg/kg bw/d and b) 0- 24.2 mg/kg bw/d; Species: a) rats and b) mice; Value Range: a) equivocal evidence of increased leukaemia incidence in female rats ≥ 7 mg/kg bw/d and b) no evidence of carcinogenic effects in male rats and in mice a. IARC (2009)

b. NTP (1992a)

Hypobromous acid Not classified No information available Relevant chemicals

Bromate a. IARC Group 2Bpossibly carcinogenic to humans
b. US EPA Group B2- probable human carcinogen based on sufficient evidence of carcinogenicity in animal a. Inadequate evidence in humans and sufficient evidence in experimental animals
b. Oral exposure in drinking water: a) 104 weeks, 0 – 500 mg/L (0 – 33 mg/kg) and b) 100 weeks, 0 – 400 mg/L (0 – 28.7 mg/kg); Species: a) and b) rats; Value Range: a) dysplastic foci ≥ 1.3 mg/kg, kidney
tumours ≥ 5.6 mg/kg, thyroid tumours, mesotheliomas at 33 mg/kg; b) mesotheliomas ≥ 6.1 mg/kg, thyroid tumours ≥ 12.9 mg/kg, kidney tumours at 28.7 mg/kg a. IARC (1999a)

b. US EPA (2001a)MEPC 60/2/9 ANNEX Page 44

I:\MEPC\60\2-9.doc Chemical Carcinogen Classification Description Reference/ Comments THMs

Chloroform a. IARC Group 2Bpossibly carcinogenic to humans

b. US EPA Group B2- probable human carcinogen based on sufficient evidence of carcinogenicity in animal a. Inadequate evidence in humans and sufficient evidence in experimental animals

b. Likely to be carcinogenic to humans by all routes of exposure under high-dose conditions that lead to cytotoxicity and regenerative hyperplasia in susceptible tissues but is not likely to be carcinogenic by any exposure route under exposure conditions that do not cause cytotoxicity and cell regeneration a. IARC (1999)

b. US EPA (2001b)

Bromoform a. IARC Group 3-not classifiable as to carcinogenicity to humans

b. US EPA Group B2- probable human carcinogen based on sufficient evidence of carcinogenicity in animal a. Inadequate evidence in humans and limited evidence in experimental animals

b. Oral exposure, gavage, 2 years, 0 – 200 mg/kg/d; Species: a) rats and b) mice; Value Range: a) increased incidence of intestinal tumours at 200 mg/kg/d and b) no neoplastic effects a. IARC (1991)

b. NTP (1989), US EPA (1991a), IPCS (2000a) and US Dept. Health (2005)MEPC 60/2/9 ANNEX Page 45

I:\MEPC\60\2-9.doc Chemical Carcinogen Classification Description Reference/ Comments

Dichlorobromomethane a. IARC Group 2Bpossibly carcinogenic to humans

b. Recommendation for EU Classification 2– Substances which should be regarded as if they are carcinogenic to man

c. US EPA Group B2- probable human carcinogen based on sufficient evidence of carcinogenicity in animal

d. Not classified by NTP a. No epidemiological evidence available; sufficient evidence in experimental animals

b. No data in humans available; conflicting results in experimental animals. Sufficient indications that carcinogenic effect may be exerted by a stochastic genotoxic mechanism

c. Oral exposure, gavage, 102 weeks: a) 0 – 100 mg/kg/d and b) 0 – 150 mg/kg/d Species: a) rats and b) mice; Value Range: a) increased incidence of kidney (100 mg/kg/d) and intestinal tumours (males dose dependent; females 100 mg/kg/d) and b) increased incidence of kidney (males, 50 mg/kg/d) and liver tumours (females, 75 and 150 mg/kg/d)

d. Oral exposure, drinking water, 2 years: a) 0/6/12/25 mg/kg/d and b) 0/9/18/36 mg/kg/d; Species: a) rats and b) mice; Value Range: a) and b) no tumours observed a. IARC (1999c);

b. Health Council of the Netherlands (2007)

c. NTP (1987), IPCS (2000a) and US EPA (1991d)

d. NTP (2006)MEPC 60/2/9 ANNEX Page 46

I:\MEPC\60\2-9.doc Chemical Carcinogen Classification Description Reference/ Comments

Chlorodibromomet hane a. IARC Group 3-not classifiable as carcinogenic to humans

b. US EPA Group C - possible human carcinogen based on limited evidence of carcinogenicity in animal a. Inadequate evidence in humans and limited evidence in experimental animals

b. Oral exposure, gavage, 2 years: a) 0/40/80 mg/kg/d and b) 0/50/100 mg/kg/d; Species: a) rats and b) mice; Value Range: a) no carcinogenic effects and b) increased incidence of liver tumours at 100 mg/kg/d a. IARC (1999b)

b. NTP (1985), US EPA (1991c) and IPCS (2000a)

HAAs

Bromochloroacetic acid a. Not Classified by IARC

b. Not Classified by NTP a. Not Classified by IARC

b. Oral exposure in drinking water, 2 years, 0 – 1000 mg/L: (a) 0 – 50 mg/kg and b) 0 – 90 mg/kg); Species: a) rats and b) mice; Value Range: Clear evidence of carcinogenic activity a) NOAEL, male: 10 mg/kg/d (malignant mesothelioma), NOAEL, female: 13 mg/kg/d (mammary gland fibroadenomas), b) LOAEL, male: 25 mg/kg/d, LOAEL, female: 15 mg/kg/d (hepatocellular adenomas) a. IARC (2009)

b. NTP (2009)

Monobromoacetic acid a. Not classified by IARC

b. Health Canada Group IV–unclassifiable with respect to carcinogenicity in humans a. Inadequate evidence for assessment of human carcinogenic potential

b. Inadequate data from animal studies a. IARC (2009)

b. Health Canada (2009)MEPC 60/2/9 ANNEX Page 47

I:\MEPC\60\2-9.doc Chemical Carcinogen Classification Description Reference/ Comments

Monochloroacetic acid a. Not classified by IARC

b. Health Canada Group IV.D–unlikely to be carcinogenic to humans a. Not Classified by IARC

b. Oral exposure, gavage, 2 years: a) 0/15/30 mg/kg and b) 0/50/100 mg/kg; Species: a) rats and b) mice; Value Range: a) and b): No evidence of carcinogenicity in mice or rats in a lifetime study; inadequate evidence for assessment of human carcinogenic potential a. IARC (2009)

b. NTP (1992b) and Health Canada (2009)

Dichloroacetic acid a. IARC Group 2B – possibly carcinogenic to humans

b. US EPA Group B2- probable human carcinogen based on sufficient evidence of carcinogenicity in animal a. Inadequate evidence in humans and sufficient evidence in experimental animals

b. Oral exposure in drinking water, 2 years, 0 – 3,500 mg/L (0 – 429 mg/kg); Species: mice; Value Range/Effects: hepatoclellular adenoma at 84 mg/kg/d and higher, clear evidence of carcinogenicity a. IARC (2004a)

b. US EPA (2003)MEPC 60/2/9 ANNEX Page 48

I:\MEPC\60\2-9.doc Chemical Carcinogen Classification Description Reference/ Comments

Dibromoacetic acid a. Not classified by IARC

b. Health Canada Group II–probably carcinogenic to humans a. Not Classified by IARC

b. Inadequate evidence in humans and sufficient evidence in animals, specifically oral exposure in drinking water, 2 years, 0 – 1,000 mg/L: a) 0 – 45 mg/kg and b) 0 – 87 mg/kg); Species: a) rats and b) mice; Value Range/Effects: a) malignant mesothelioma at 40 mg/kg/d (males), mononuclear cell leukemia at 45 mg/kg/d (females) and b) liver
tumours ≥ 4 mg/kg/d (males), ≥ 35 mg/kg/d (females), lung tumours ≥ 45 mg/kg/d (males) a. IARC (2009)

b. NTP (2007) and Health Canada (2009)

Trichloroacetic acid a. IARC Group-3-not classifiable as carcinogenic to humans

b. Health Canada Group III–possibly carcinogenic to humans

c. US EPA Group C - possible human carcinogen based on limited evidence of carcinogenicity in animal a. Inadequate evidence in humans and limited evidence in experimental animals

b. Inadequate evidence in humans and limited evidence (one species) in animals

c. Oral exposure in drinking water, 2 years, 0- 5,000 mg/L; a) 0 – 284 mg/kg and b) 0 – 595 mg/kg); Species: a) rats and b) mice; Effects/Value Range: a) no carcinogenic effects and b) dose-dependent increase in liver tumours > 71 mg/kg/d a. IARC (2004b )

b. Health Canada (2009)

c. US EPA (1996) and IPCS (2000a)MEPC 60/2/9 ANNEX Page 49

I:\MEPC\60\2-9.doc Chemical Carcinogen Classification Description Reference/ Comments Other Chemicals

Sodium bisulfite a. IARC Group 3-not classifiable as carcinogenic to humans

b. American Conference of Governmental Industrial Hygienists TLVs and BEIs Group A4-not classifiable as a human carcinogen a. Inadequate evidence for carcinogenicity in humans for sulfur compounds and limited evidence in experimental animals. IARC 1992 and IPCS (2000a)

4.7 Mutagenicity/genotoxicity

Table 14 below summarizes data on mutagenicity and genotoxicity for the chemicals involved in STDN’s BalPure ® treatment system.

Table 14 : Summary of Data on Mutagenicity and Genotoxicity

Chemical Study Results Reference/ Comments Active Substances

Hypochlorous acid Some studies show that sodium chlorite causes mutagenic responses in b acteria and mammalian cells in vitro . There is no evidence of mutagenic response in mammalian systems in vivo. IPCS (2000a)

Hypobromous acid No information available Relevant Chemicals

Bromate Some studies show positiv e results for genotoxic activity in several in vitro and in vivo systems. IPCS (2000a), IARC (1999a), US EPA (2001a) THMs

Chloroform Studies show negative results for mutagenicity in several in vitro and in vivo assays. IPCS (2000a)

Bromoform Studies show positive results for mutagenicity in several in vitro and in vivo assays. In vivo studies with rodents bromoform did not cause chromosomal aberrations IARC (1991), IPCS (2000a)

Dichlorobromomethane Increased frequency of sister chromatid exchange in mice dosed with 50 or 100 mg/kg/day for 4 days. ATSDR & US EPA (1989)

Chlorodibromomethane Studies show positive results for mutagenicity in several bacterial and eukaryotic in vitro systems; equivocal results are reported from in vivo studies in rodents. IPCS (2000a)MEPC 60/2/9 ANNEX Page 50

I:\MEPC\60\2-9.doc Chemical Study Results Reference/ Comments HAAs

Bromochloroacetic acid Some positive results for Salmonella typhimurim for genotoxicity. Negative genotoxic results for male and female mice exposed to BCAA for 3 months in drinking water. HSDB/TOXNET (2009b)

Monochloroacetic acid Data is conflicting; most evidence suggests no significant mutagenic potential WHO (2004b)

Monobromoacetic acid Mutagenic in Salmonella typhimurium . Chromosomal damage not observed in newt micronucleus test. WHO (2004a)

Dichloroacetic acid Conflicting results are reporte d from several in vitro and in vivo studies; the overa ll dataset indicated that DCA induces mutations and chromosomal effects. IARC (2004a)

Dibromoacetic acid Studies show positive results for mutagenicity in several in vitro and in vivo assays. IPCS (2000a)

Trichloroacetic acid Weakly mutagenic. Did not induce chromosomal aberrations or DNA strand breaks in single studies, and no effect observed in a DNA strand-break assay or single-cell gel assay. IARC (2004b)

Other Chemicals

Sodium bisulfite Not mutagenic in rats at doses up to 150 mg/kg. No mutagenic activity in human tissue cells in vitro up to 200 μg/mL. WHO/IPCS (2008)

4.8 Toxicokinetics

Table 15 below characterizes the existing data concerning the uptake, absorption, and excretion of the chemicals.

Table 15 : Summary of Uptake, Absorption, and Excretion of Chemicals

Chemical Study Results Reference/ comments Active Substances Hypochlorous acid Hypochlorous acid breaks down into several different chemicals in the gut such as trichloroacetic acid, dichloroacetic acid, a nd dichloroacetonitrile HSDB/TOXNET (2009i)
Hypobromous acid No data found in literature Relevant chemicals

Bromate (as sodium bromate) After a dose of 230-460 mg BrO 3 - /kg in a 2 year old child (13 kg), 60-70% was r ecovered from dialysate and urine as bromide (1,850mg). Bromate is rapidly absorbed from gastrointestin al tract, partially converted to bromide in tissues and excreted rapidly HSDB/TOXNET (2009k) Health Canada (1998)MEPC 60/2/9 ANNEX Page 51

I:\MEPC\60\2-9.doc Chemical Study Results Reference/ comments THMs

Chloroform Well absorbed, metabolized, and eliminated rapidly by mammals after oral, inhalation, or dermal exposure. After dose of 0.5 g chloroform, 50-52% of dose absorbed and approx. 100% of dose metabolized to carbon dioxide IPCS (1994)

Bromoform After administered to mice and rats, 3-6% and 5-14% remains in body tissues after 36 or 48 hrs, respectively. HSDB/TOXNET (2009l)

Dichlorobromom ethane Moves from stomach/intestines into the blood. Leaves body mostly through exhalation and less through urine and feces. Removal fairly rapid (95% in 8 hours) ATSDR & US EPA (1989)

Chlorodibromom ethane Rats administered with 100 mg/kg bw absorbed and eliminated as expired air as BDCM or CO 2 in 8 hours. 1% eliminated in urine and 1% retained in body tissues. Metabolized to CO in vitro and in vivo HSDB/TOXNET (2009e)

HAAs

Bromochloroacetic acid Rapidly cleared from blood following single intravenous dose. Urine and feces minimal contributors-suggests elimination via conversion to CO 2 WHO (2004b)

Monochloroacetic acid In mice, 80-90% of administered MCA excreted in urine within 24 hours; 8 % was exhaled as CO 2 IPCS (2000b)

Monobromoacetic acid No studies have been found in literature WHO (2004a)

Dichloroacetic acid Both humans and rodents metabolize DCA to glyoxylate by oxidative de-chlorination; the plasma half-life of the parent compound Is 0.5-2 hours HSDB/TOXNET (2009h)

Dibromoacetic acid Limited in vitro data suggests rapid excretion and minimal bioaccumulation. Urine and feces minimal contributors to blood cl earance, suggesting biotransformation WHO (2004a)

Trichloroacetic acid After administration of single or al dose of 3 mg/kg bw to volunteers, mean plasma ha lf life was ~50 hrs and volume of distribution was ~115 mL/kg bw. TCA is absorbed rapidly and expelled in urine as TCA or oxalic/thiodiacetic acids HSDB/TOXNET (2009m)

Other Chemicals

Sodium bisulfite Rats given a 0.2% solution eliminated 55% of administered sulfur as sulfate in urine within first four hours. Rapid elimination of sulfites as sulfate observed in man and dog. WHO /IPCS (2008)MEPC 60/2/9 ANNEX Page 52

I:\MEPC\60\2-9.doc 5 DATA ON ENVIRONMENTAL FATE AND EFFECT UNDER AEROBIC AND ANAEROBIC CONDITIONS (G9: 4.2.1.3)

5.1 Modes of degradation (biotic; abiotic)

As shown in Table 16 below, the Active Subs tances (hypochlorous and hypobromous acid) decay in seconds to hours; this is after all their mechanism of ac tion and how they produce an oxidizing environment. As a consequence of this oxidi zing environment, along with the presence of organic matter, THMs and HAAs are also produ ced as described in section 2.3. For THMs and HAAs, both the biotic and ab iotic degradation data are shown in Table 16. Biotic degradation for all of these simple halogenate d organic compounds is on the order of days to weeks. Abiotic degradation of these compounds is longer.

5.2 Bioaccumulation, partition coefficient, octanol/water partition coefficient

The bioconcentration factor (BCF) is the concentr ation of a particular chemical in a tissue per concentration of chemical in water (reported as L/kg). This phys ical property characterizes the accumulation of pollutants through chemical parti tioning from the aqueous phase into an organic phase, such as the gill of a fish. Generally, a hi gh potential BCF is greater than 1000, a moderate potential BCF is less than a 1000 but greater th an 250, and a low potential BCF is less than 250. The literature reported and/or estimated BCFs are shown for the Active Substances and Relevant Chemicals in Table 7.

The organic carbon adsorption coefficient (Koc) is the chemical adsorption coefficient normalized to the organic carbon content of soil or sediment. This parameter describes the tendency of a substance to bind to orga nic matter in the soil or sediment.

The octanol/water partition coefficient (Kow) is defined as the ratio of a chemical’s concentration in the octanol phase to its con centration in the aqueous phase of a two-phase octanol/water system. Measured values of log Ko w for organic chemicals range from -3 to 7. Generally, chemicals with low Log Kow valu es can be considered relatively hydrophilic (i.e. tendency for high water solubility, low pote ntial for sediment adsorption), while chemicals with high Log Kow values are considered hydr ophobic (i.e. low water sol ubility, high potential for sediment adsorption). The literature repo rted and/or estimated log Kows are shown for the Active Substances and Releva nt Chemicals in Table 16.

Based on the data shown in Table 7 for both Activ e Substances and Relevant Chemicals all of these substances have very low BCFs, indicati ng that they are unlikely to bioaccumulate in aquatic organisms or present a significant food ch ain risk. With the exception of the THMs, all of the Active Substances and Relevant Chemicals have very low log Kow’s, which indicates the compounds are hydrophilic. The THMs, as woul d be expected, are m oderately hydrophobic. However, none of the log Kow values located in the literature and/or estimated are ≥ 3. The log Kow and BCF are related to each other w ith the BCF being the more relevant parameter to determine the likelihood of impa cts of a chemical to aquatic organisms. The Koc’s for all chemicals in Table 16 are all at or below 250 in dicating a low likelihood of partitioning into the sediment compartment of aquatic systems.MEPC 60/2/9 ANNEX Page 53

I:\MEPC\60\2-9.doc Table 16 : Modes of Degradation

Chemical Information on Modes of Degradation BCF/Koc/ Log Kow Comments/ References Active Substances

Hypochlorous acid Decays within seconds to hours, no rate specified BCF: 3.2 Koc: 13.2 Log Kow: -0.87 Echigo & Minear (2006)/EPI Suite (2009c)

Hypobromous acid Decays within seconds to hours, no rate specified BCF: 3.2 Koc: 13.2 Log Kow: -0.78 Echigo & Minear (2006)/EPI Suite (2009b) Relevant Chemicals Bromate (as sodium bromate) No information located. BCF:3.2 Koc: 31.8 Log Kow: -4.6 HSDB/ TOXNET (2009k)/EPI Suite (2009a) THMs

Chloroform BIOTIC: Estimated volatilization half-lives for model river and lake are 1.3 hrs and 4.4 days, respectively. No marine biodegradation reported. Can be biodegraded under anaerobic conditions by methanogenic bacteria. ABIOTIC: Atmospheric ha lf life of ~151 days at atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3 . Avg. degradation rate of 0.8%/hr. BCF: 2.9-10.35 Koc:153- 196 HSDB/ TOXNET (2009f)

Bromoform BIOTIC: In anoxic groundwater environments: 20% degradation after 6 hours, 99% degradation after ~2.5 days. Volatiliza tion from water surface expected. Estimated volatil ization half lives for model river and lake are 7.3 hours and 7.1 days, respectively. ABIOTIC: Atmospheric half life of ~325 days at an atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3 . Hydrolysis is not expected. BCF: 14 Koc: 35 Log Kow: 2.4 HSDB/ TOXNET (2009l)

Chlorodibro mo-methane BIOTIC: Volatilization fr om water surfaces is expected. Volatilization ha lf-lives for model river and lake are 2.6 hrs and 6 days, respectively. Resistant to aerobic biod egradation; degrades anaerobically within 2 to 8 weeks. ABIOTIC: Atmospheric ha lf life of ~280 days at an atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3 . Direct photolysis or aquatic oxidation not relevant processes. BCF: 9 Koc: 84 HSDB/ TOXNET (2009e)MEPC 60/2/9 ANNEX Page 54

I:\MEPC\60\2-9.doc

Chemical Information on Modes of Degradation BCF/Koc/ Log Kow Comments/ References

Dichlorobro mo-methane BIOTIC: 51-59% biodegradation over 28 days in aerobic conditions using static flask screening procedures. 100% biodegr adation in anaerobic conditions after 2 weeks using methanogenic bacteria from sewage. (No degradation in control without bacteria). ABIOTIC: Atmospheric ha lf life of 205 days at atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3 . Direct photolysis not expected. Aqueous hydrolysis half life estimated at 137 years (25  ̊ C and pH 7) BCF: 7 Koc: 35- 251 Log Kow: 2 HSDB/ TOXNET (2009c)

HAAs

Bromochloroacetic acid BIOTIC: Volatilization fr om water surfaces not expected. (No biodegrada tion data available) ABIOTIC: Atmospheric ha lf life of ~23.8 days at atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3 . Not expected to undergo hydrolysis or direct photolysis by sunlight. BCF: 3.2 Koc: 1.9 Log Kow: 0.61 HSDB/ TOXNET (2009b)

Monochloroacetic acid BIOTIC: Volatilization fr om water surfaces not expected. In aerobic conditions CAA reaches 65% of theoretical BOD in 3 weeks at 100 mg/L. No biodegradability in seawat er after 3 days using cultivation method at 10 mg/L. Difficult to degrade. ABIOTIC: Atmospheric half life of ~ 20 days at atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3 . Hydrolysis and direct photolysis by sunlight not expected. BCF: 3.2 Koc: 31 Log Kow: 0.22 HSDB/ TOXNET (2009d)

Monobromoacetic acid BIOTIC: Volatilization fr om water surfaces not expected. In biodegradat ion screening tests, reached 82% of theoretic BOD when using acclimated inoculums. (not readily biodegradable w/out un-acclimated inoculums). ABIOTIC: Atmospheric half life of ~22.4 days at an atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3 . Not expected to undergo hydrolysis or direct photolysis from sunlight. BCF: 3.2 Koc: 39.8 Log Kow: 0.41 HSDB/ TOXNET (2009a)

Dichloroacetic acid BIOTIC: Biodegradability reported to be 14 and 8% after 3 days of incubation for river and seawater, respectively. Volatilization from water is not expected due to pKa values. ABIOTIC: Atmospheric half life of ~22 days at atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3 . Not expected to undergo hydrolysis or direct photolysis by sunlight. BCF: 0.3 Koc: 75 Log Kow: 0.92 HSDB/ TOXNET (2009h)MEPC 60/2/9 ANNEX Page 55

I:\MEPC\60\2-9.doc

Chemical Information on Modes of Degradation BCF/Koc/ Log Kow Comments/ References

Dibromoacetic acid BIOTIC: Limited data available. Volatilization from water surface is not expected due to pKa values. ABIOTIC: Atmospheric half life of ~ 25.3 days at atmospheric concentration of 5X10 5 hydroxyls radicals/cm 3 . Not expected to undergo hydrolysis or direct photolysis by sunlight. BCF: 0.17 Koc: 1.5 HSDB/ TOXNET (2009g)

Trichloroacetic acid BIOTIC: Volatilization from water surfaces is not expected due to pKa values. At concentrations of 100 mg/L reaches 7% of theoretical BOD in 4 weeks using activated sludge. ABIOTIC: Atmospheric ha lf life of ~31 days at atmospheric concentration of 5X10 5 hydroxyl radicals/cm 3

. BCF: 0.1- 1.7 Koc: 130 Log Kow: 1.33 HSDB/ TOXNET (2009m)

Other Chemicals

Sodium bisulfite No information located. BCF: 3.2 Koc: 2.21 Log Kow: -6.85 EPI Suite (2009d)

5.3 Persistence and identification of the main metabolites in the relevant media

Higher tier simulation tests not re quired; section not applicable.

5.4 Reaction with organic matter

The formation of disinfection by-products (DBPs) when using chlorine-bas ed water disinfection methods is well known. Two major classes of DB Ps that are commonly formed in chlorine treated water include THMs and HAAs (Westerhoff, et al., 2003). Both of these DBP classes have been measured in water treated with the BalPure ® system. DBP formation is a result of disinfectant reactions with natural organic matter, often meas ured as total organic carbon (TOC), which serves as the organic precursor, and/or bromide ion, which serves as the inorganic precursor (IPCS, 2000a). Reactions between disi nfectant halogens (e.g., chlorine and bromine) and DBP precursors include substitution reac tions and carbon bond oxidation reactions (Westerhoff, et al., 2003). Water quality (e.g., pH, temp erature, bromide level, TOC) and treatment conditions (e.g., hypochlor ite dose, contact time, remova l of organic matter prior to treatment) influences the formation of DBPs. Further, the distributio n of DBP species (four THM species, up to nine HAA species) is affected by the amounts of TOC, bromide and chlorine present (IPCS, 2000a).

Analysis for DBP concentrations has been conducted during pilo t scale testing of the BalPure ® system. Different water types (freshwater, brac kish, seawater) were treated and then evaluated for DBPs. The data are presented in sectio n 11 “Evaluation of Treated Ballast Water”. Additional evaluation of DBP concentrations in treated ballast water will be performed during land-based testing at NIOZ and the results will be reported in the Final Approval dossier.MEPC 60/2/9 ANNEX Page 56

I:\MEPC\60\2-9.doc 5.5 Potential physical effects on wildlife and benthic habitats

When evaluated using GESAMP Reports and Studies No. 64, the chemical and physical properties of the Active Substances and Releva nt Chemicals associated with the BalPure ® system (Tables 17, 18, 19, 20, and 21) indicate that physical effects on wild life and benthic habitats as described are not expected to occur.

5.6 Potential residues in seafood

Chemicals detected in treated ballast water that is discharged may pose a threat to seafood consumers only if they tend to bi oaccumulate (within a trophic leve l, an increase in concentration of a substance in tissues due to uptake from food and sediments) or biomagnify (an increase in the concentration of a substance in the fo od web). The bioaccumulation criteria per the Methodology for information gathering and the conduct of work of the GESAMP-BWWG (MEPC 58/2/7, annex 4) include an octanol-water partition coefficient (Log K ow ) of ≥ 3 or a bioconcentration factor (BCF) >2000. The chemicals identified in this risk assessment all have Log K ow s less than 3 and BCFs less than 2000, and th erefore are not expected to persist in the food web.

5.7 Any known interactive effects

No other interactive effects are known.

6 PHYSICAL AND CHEMICAL PROPERTI ES OF THE ACTIVE SUBSTANCES, RELEVANT CHEMICALS AND TREA TED BALLAST WATER (G9: 4.2.1.4)

Table 17 : Physical and Chemical Properties of Active Substances

Physical/Chemical Property Active Substance

− Hypochlorous Acid Active Substance

− Hyprobromous Acid Melting point (o C) 194.89oC 208.57oC Boiling point (o C) 507.1oC 527.32oC Flammability (flash point for liquids; oC) Not flammable Not flammable
Density (20oC; kg/m3) 1.21 Not located. Vapor pressure/Vapor density (air=1) 1.85 x10 -12 mm Hg 3.06 x10 -13 mm Hg Water solubility (temp; effect of pH; mg/L)/ Dissociation constant (pKa) 1x10 6 (25oC) 1x10 6 mg/L (25oC)

Oxidation/Reduction potential 700-800 mv* 700-800 mv*
Corrosivity to materials Corrosive Data not located Auto-ignition temperature (oC) Data not located Data not located

Explosive properties In ballast water – not applicable In ballast water – not applicable Oxidizing properties Strong oxidizing agent Oxidizing agent Surface tension Same as ballast water Same as ballast water Viscosity (20oC) Same as ball ast water Same as ballast waterMEPC 60/2/9 ANNEX Page 57

I:\MEPC\60\2-9.doc Physical/Chemical Property Active Substance

− Hypochlorous Acid Active Substance

− Hyprobromous Acid

Thermal stability Very stable in aqueous solutions between 15- 18oC Data not located

Reactivity to container material Not stored; produced in situ Not stored; produced in situ

Other known’s May have slight chlorine-like odor None

*ORP as measured in ballast wate r after treatment with hypochlorite. Note : All data from HSDB/TOXNET unless otherwise specified.

Table 18 : Physical and Chemical Pr operties of Relevant Chemicals-THMs

Physical/Chemical Property THMs Chloroform THMs Bromoform THMs Chlorodibrom

omethane THMs Dichlorobromo -methane

Flammability (flash point for liquids; oC) Non-flammable Non-flammable Non-flammable (US Dept. of Health and Human Services and ATDSR, 2005) Non-flammable

Vapor pressure/Vapor density (air=1) 197 mmHg (25oC) 1 Kpa (30  ̊ C)/8.7 5.54 mm Hg (25oC) 50 mm Hg (20oC)
Water solubility (temp; effect of pH; mg/L)/ Dissociation constant (pKa) 7,710 mg/L (25oC) 3,100 mg/L (25oC) 2,700 mg/L (20oC) 4,700 mg/L (22oC)

Oxidation/Reduction potential 700-800 mv* 700-800 mv* 700-800 mv* 700-800 mv*

Corrosivity to materials Aqueous form corrosive to some forms of plastics, rubber, and coatings (CCOHS, 2007) Aqueous form corrosive to steel and gray cast iron, and some plastics (CCOHS, 2007) Data not located Data not located

Auto-ignition temperature (oC) NA NA NA (US Dept. of Health and Human Services and ATSDR, 2005) NAMEPC 60/2/9 ANNEX Page 58

I:\MEPC\60\2-9.doc Physical/Chemical Property THMs Chloroform THMs Bromoform THMs Chlorodibrom

omethane THMs Dichlorobromo -methane

Explosive properties Explosive in presence of some alkali metals and strong mineral acids Explosive in presence of crown ethers or potassium hydroxide Incompatible with strong bases, strong oxidizing agents, and magnesium Incompatible with strong bases and strong mineral acids

Oxidizing properties Strong oxidizing agent No data available No data available No data available
Reactivity to container material Not stored; produced in situ Not stored; produced in situ Not stored; produced in situ Not stored; produced in situ

Other known’s None None None None NA=Not applicable *ORP as measured in ballast wate r after treatment with hypochlorite.

Note : All data from HSDB/TOXN ET unless specified otherwise

Table 19 : Physical and Chemical Pr operties of Relevant Chemicals-HAAs

Physical/ Chemical Property HAAsBromochloroacetic acid HAAsMonochloroacetic acid HAAsMonobromoacetic acid HAAsDichloroacetic acid HAAsDibromoacetic

acid HAAsTrichloroacetic acid Flammability (flash point for liquids; oC) 110oC (closed cup) 126oC (closed cup) 113oC (closed cup) 113oC 113oC (closed cup) 198oC

Vapor pressure/ Vapor density (air=1) 0.14 mm Hg (25oC) 0.065 mmHg (25oC)/3.26 0.119 mm Hg (25oC) 0.179 mm Hg (25oC)/4.45 0.023 mm Hg (25oC) 0.06 mm Hg (25oC)

Water solubility (temp; effect of pH; mg/L)/ Dissociation constant (pKa) 2.5 X10 5 mg/L (25oC)/ pka=1.4 8.5810 5 mg/ L(25oC)/ Pka=2.87 1.75X10 6 mg /L(25oC)/ pka=2.89 Miscible with water/ pka=1.26 2.1X10 6 mg/L (25oC)/ pka=1.48 5.4X10 4 mg/L (25oC)/ pka=0.51

Oxidation/ Reduction potential 700-800 mv* 700-800 mv* 700-800 mv* 700-800 mv* 700-800 mv* 700-800 mv*

Corrosivity to materials Data not located Data not located Data not located Data not located Data not located Corrosive to iron, zinc, aluminiumMEPC 60/2/9 ANNEX Page 59

I:\MEPC\60\2-9.doc Physical/ Chemical Property HAAsBromochloroacetic acid HAAsMonochloroacetic acid HAAsMonobromoacetic acid HAAsDichloroacetic acid HAAsDibromoacetic

acid HAAsTrichloroacetic acid Auto-ignition temperature (oC) Data not located >500oC Data not located Data not located Data not located Data not located

Explosive properties Incompatible with strong oxidizing agents Explosive in presence of strong oxidizers, strong bases, and strong reducing agents Closed containers may explode when heated

or contaminated with water) Vapors in confined
area may explode in fire. Data not located Data not located

Oxidizing properties Data not located Data not located Data not located Data not located Data not located Data not located
Reactivity to container material Not stored; produced in situ Not stored; produced in situ Not stored; produced in situ Not stored; produced in situ Not stored; produced in situ Not stored; produced in situ

Other known’s None None None None None None *ORP as measured in ballast wate r after treatment with hypochlorite. Note : All data from HSDB/TOXNET unless otherwise specified.

Table 20 : Physical and Chemical Properties of Relevant Chemicals and Other Chemicals

Relevant Chemicals Other Chemicals Physical/Chemical Property Bromate Hydrogen Gas Sodium Bisulfite Flammability (flash point for liquids; oC) Data not located Extremely flammable (ICSC) Non-flammable

Vapor pressure/Vapor density (air=1) Data not located 0.07 (ICSC) 1.48

Water solubility (temp; effect of pH; mg/L)/Dissociation constant (pKa) 3.64 g/L (20oC) Not applicable Very soluble in
water

Oxidation/Reduction potential 700-800 mv* 700-800 mv* 700-800 mv*

Corrosivity to materials Data not located NA Data not located Auto-ignition temperature (oC) Data not located 500-571  ̊ C (ICSC) Data not locatedMEPC 60/2/9 ANNEX Page 60

I:\MEPC\60\2-9.doc Explosive properties Prolonged exposure to heat may result in explosion Gas/air mixtures are explosive; LEL: 4-76% volume in air (ICSC) Not considered explosive

Oxidizing properties Oxidizing agent Not Applicable None Reactivity to container material Not stored; produced in situ Not stored; produced in situ Not stored; produced in situ Other known’s None None None

NA=Not applicable *ORP as measured in ballast wate r after treatment with hypochlorite. Note : All data from HSDB/TOXNET unless otherwise specified.

Table 21 : Physical and Chemical Properties of Treated Ballast Water

Physical/Chemical Property Treated Ballast Water Density (20oC; kg/m3) Same as seawater Oxidation/Reduction potential <200 mV*

Corrosivity to materials Similar to untreated ballast
water

Reactivity to container material Similar to untreated ballast
water pH See Table 26 Salinity See Table 26 TOC See Table 26 DOC See Table 26 Other known’s None *ORP as measured in ballast water after neutra lization with sodium bisulfite.

7 ANALYTICAL METHODS AT ENVIRONMENTALLY RELEVANT CONCENTRATIONS (G9: 4.2.1.5)

7.1 Analysis of Total Residual Oxidant (TRO)

TRO is measured at two concentration ranges (high and low) using tw o different analytical procedures. An iodometric ti tration procedure is used to measure higher range TRO in the product stream exiting the BalPure ® unit. Concentrations are typically 1,000 mg/L and this procedure is accepted by the American Water Works Association (AWWA) for examination of water and wastewater. The proc edure is only used if needed to perform diagnostics on the BalPure ® system, and is incl uded in appendix A.5 3 . A second method is typically used for measuring lower range TRO in potable water and waste water streams. A Hach colorimeter can be used to measure the TRO in ballast tanks and in the de-ballast stream prior to neutralization. The TRO oxidizes triiodide ion (I 3 -) to iodine (I 2 ). The iodine and free chlorine react with DPD (N,N-diethyl-p-phenylenediamine) to form a red solution. The color intensity is proportional to the TRO concentration. This method is also provided in appendix A.5 3

.

3 Appendix A.5 is provided in the confidential dossier of the proposal for approval.MEPC 60/2/9 ANNEX Page 61

I:\MEPC\60\2-9.doc During system testing at the University of Washington, TRO measurements were performed internally by the researchers using a Hach DR/8 20 colorimeter and the Hach DPD powder pillow Method 10070 in appendix A.5 1 (equivalent to EPA Method 330.5).

7.2 Analysis of Disinfecti on By-Products (DBPs)

During the pilot scale st udies of the BalPure ® system, qualified and accr edited laboratories were utilized for analysis of treated water sa mples. The methods used to determine DBP concentrations are listed in Table 22 below.

Table 22 : Analytical Methods

Chemical/ Parameter US EPA Method Description Method Detection Limit (MDL)

TRO (Range: 0.1-10 mg/L) Hach Method 10070 (Equivalent to EPA Method 330.5) DPD, colorimetric 0.1 mg/L

THMs 524.2 Capillary column gas chromatography/mass spectrometry 0.02 - 1.6
μ g/L*

HAAs 552.1 Ion-exchange liquid-solid extraction and gas chromatography with an electron capture detector 0.07 – 0.45
μ g/L*

Bromate 300.1 Ion chromatography 1.28 – 1.44
μ g/L* *MDL as determined by the analytical method. MDL is compound, instrument and especially matrix dependent.

8 USE OF ACTIVE SUBSTANCE

8.1 The manner of application

The BalPure ® system can service ships with ballast water flow requirements from 100 to 10,000 cubic metres per hour. The BalPure ® system size (electrolytic cell si ze) is defined by the ship’s designed ballast water flow rate, but because th e system has a 100% turn down capability the hypochlorite production rate is matched to the actual measured ballast flow rate. Treatment is done during ballasting operations allowing the bioc ide solution to be generated anywhere on the ship and injected as needed. This would apply to ships with one or multiple ballast systems or ships with multiple ballast pumps mounted within separate ballast tanks. The system can be delivered as a complete system on one skid or divided into seven separate smaller skids based on specific unit operation (pumpi ng, hypochlorite generation, hydroge n-hypochlorite separation, bisulfite storage, bisulfite mete red addition, instrument and contro ls, and power re ctification). This allows multiple skid locations as space is available throughout the vessel. The multiple skid design is perfect for retrofit applications and the one skid system is best for new builds.MEPC 60/2/9 ANNEX Page 62

I:\MEPC\60\2-9.doc Operational control of the BalPure ® system is accomplished by the main control panel or by the remote panel located in the cargo control room. Together, the control panel and the transformer/rectifier unit comprise the main Programmable Logic Contro ller (PLC) control and rectification components. The control panel pr ovides a visual display of the status of the rectifier DC voltage, current, a nd can alert the operator of any system alarms via the operator interface terminal (OIT). The main control pane l also has a local emergency stop push button. All major equipment such as booster pumps, bisulfite metering pumps, and hydrogen blowers can be controlled from the main control panel. All monitoring equipment such as flow meters and ORP probes send the out put in the form of 4-20 mA signals . All system control screens are password protected to limit operator modification to existing control logic. Only properly trained ship’s crew members and BalPure ® technicians will be allowed to access password-protected portions of the PLC.

Other system monitoring and control devices in clude a magnetic flow transmitter to monitor the flow of seawater at the electr olyzer inlet, transformer temper ature switches that monitor the internal temperature of the rec tifier, and airflow switches that monitor the hydrogen gas dilution blowers.

8.1.1 Process flow description

The BalPure ® system is designed to operate au tomatically and has two modes of operation: ballasting and de-bal lasting. During ballasting, the bi ocide solution is produced and injected into the ballast water intake piping; during de-ballasting sodium bisulfite is added directly to the ballast water disc harge piping. The ballasting mode is discussed firs t, followed by the de-ballasting mode. A complete process flow diagram (PFD) of the system is included in appendix A.3 4 . Figures 5 and 6 presented below provide a simplified overview of each phase of the treatment process.

Very generally, during the automatic ballasting mode, the system will sense that adequate seawater flow is available using a flow meter in the main ballast line. Once the system is primed with sufficient water (an approximate 1 minute cycle), the transformer/rectifier will supply DC current to the electrolyzer and initiate the electrochemical generation process. The hypochlorite production rate is pr imarily tied to the ballast flow rate and achieves the pre-set required oxidant concentration by m odulating the rectifier output am perage to coincide with the ballast flow rate. This is considered the larg e or gross adjustment. The produced oxidants are then injected into the main ballast line. If n ecessary, the system may also be operated in manual mode. Figure 5 below provides an overview of the BalPure ® ballasting process.

4 Appendix A.3 is provided in the confidential dossier of the proposal for approval.MEPC 60/2/9 ANNEX Page 63

I:\MEPC\60\2-9.doc Figure 5 : Overview of the BalPure ® Ballasting Process

The BalPure ® system remains in a stand-by mode until the flow meter in the main ballast line detects ballast water flow and activates the BalPure ® system. In the ballasting mode, seawater from the sea chest will be filtered with a 40 micr on stainless steel mesh filtration unit. If an installation requires that seawater be taken from a source other than filtered ballast water (e.g., seawater cooling), a duplex strainer filters the seawater to remove any particles larger than 800 microns. Using an inline booster pump, the BalPure ® system typically receives <1% of the total incoming ballast water flow as a side-str eam off the main ballast intake line. An inlet flow transmitter, designed to protect the treatment system, will shut down the electrical current to the electrolyzer should seawater flow through th e electrolyzer drop below 80% of the design flow rate. The filtered seawater passes through a he at sink where it absorbs excess heat generated by the silicon-controlled rectifiers . The seawater then passes through a series of electrolytic cells, which are connected to the transformer/rectif ier. The electrolyzer units then generate the Active Substance along with hydrogen gas as a by-product.

Immediately after exit from the electrolytic cells, the combined seawater/biocide solution (liquid) and hydrogen (gas) is delivered to a patented cyclone separation device where the hydrogen is separated from the biocide/seaw ater solution. Hydrogen gas doe s not enter the ballast water piping or tanks. The hydrogen gas is directed to a vent stack where it is diluted by forced air blowers and vented to the atmosphere outside of the ship. To ensure pr oper ventilation, the vent line is serviced by two redundant blowers couple d to sail switches. The sail switches ensure proper air flow to dilute the air stream to <1% hydrogen. The LEL for hydrogen is 4% and the operating limitations on the system provide a fou r-fold safety factor. The PLC monitors proper air flow and will shut down the system, resulting in an alarm condition if proper ventilation air flow is not maintained. Both blower motors and sail switches are tied to an emergency shutdownMEPC 60/2/9 ANNEX Page 64

I:\MEPC\60\2-9.doc for the production unit in the unlikely event all four devices fail. For additional safety, a hydrogen sensor with alarm will be pr ovided in the general work area.

For optimum hypochlorite generator performance (i.e. to produce 90% of theoretical values), the inlet seawater temperatur e should be at least 15 ° C and have a chloride concentration of at 19,000 mg/L (19 PSU). However, the BalPure ® system can operate at temperatures as low as 5 ° C and salinities of 10,000 mg/L (10 PSU). Less optimal conditions result in a slightly decreased hypochlorite production rate to 20% less than optimu m. The hypochlorite production rate of the electrolytic cells is directly rela ted to chloride ion av ailability and mobility. The salinity (PSU) of the source water determines the chloride ion availability, while temperature affects chloride ion mobility. These parameters impact the operating volta ge of the electrolytic cells. For instance, when chloride ions are abundantly available and mobile, the operating voltage and hypochlorite production w ithin the electrolytic cells wi ll be optimum. If chloride ions and/or temperature are low, the operating voltage increase s proportionally and hypochlorite production will be reduced. The BalPure ® system is designed to adjust the hypochlorite production rate by monitoring the operating voltage of the electrolytic cel ls and increasing the rectifier operating amperage to sustain the TRO concentration required to treat incoming ballast water. These adjustments are small and used to control the hypochlor ite production rate to a narrow range (fine tuning).

The BalPure® system can also operate at higher salinities, such as ≥ 30,000 mg/L ( ≥ 30 PSU). In this case where chloride i ons are highly abundant, the gene rator will only produce the design hypochlorite concentration. This is because higher salinity does not increase the efficiency or the production rate of the electro lytic cell beyond the optimum.

By monitoring the operating voltage of the electrolytic cells and adjusting the amperage applied, the PLC programming of the BalPure® system guarantees effective operation over a wide temperature and/or PSU range.

In the de-ballast mode of operation, the last ph ase of the treatment process, residual chlorine present in the ballast tank is neutralized (de-chlo rinated) with sodium bisulfite just prior to discharge from the ship. This is accomplished using ORP tec hnology and bisulf ite metering pumps. Two ORP probes are used; one confirms the presence of residual oxidant exiting the ballast tanks, while the s econd probe monitors and controls the addition of sodium bisulfite to neutralize TRO during ballast wate r discharge. The first ORP probe is located upstream of the bisulfite injection point, and the second ORP probe is located downstream of the bisulfite injection point.MEPC 60/2/9 ANNEX Page 65

I:\MEPC\60\2-9.doc Figure 6 : Overview of the BalPure® De-ballasting Process

ORP technology is based on the potential measur ement between two dissimilar metals within the ORP probe. ORP is not a quantitative measure of oxidant or bisulfite concentration but rather a qualitative measure that confirms the presence of oxidizing or reducing agents in the water. ORP technology allows for an in-line rapid de termination of the am ount of oxidation (or reduction) potential of the water. ORP is measured in millivolts (mV) and increases or decreases in direct relation with the oxi dant residual in the water (Ke lley, 2004). For example, natural waters have an ORP reading in the range of 100 – 150 mV. When hypochlorit e is added to water, the ORP reading increases with an increas e in hypochlorite dosage. Similarly, when a de-chlorinating agent such as sodium bisulfite is added to the water, the ORP reading will drop to a low mV level (Kelley, 2004). In this way, ORP measurements pr ovide a practical and efficient method of optimizing water disinfection a nd controlling neutralizing agent dosage.

The first BalPure® ORP probe confirms that ther e are residual oxidants in the treated water being discharged from the ballast tank. In principle, this confirma tion of oxidants translates to affirmation that there are no livi ng organisms in the discharge. Th e ORP reading at the first probe is typically between 700 and 800 mV. The second ORP probe measures the ORP value of the de-ballast stream after bisulfite has been a dded to the discharge piping. This ORP value is typically <200 mV and is used to control the addition of bisulfite solution to ensure complete neutralization of re sidual oxidants. With the mixing effect of the de-ballast water pump and the discharge pipe, the bisulfite will easily react with any free oxidants that may be present. As a fail-safe measure, the BalPure® system is f itted with two redundant bisulfite metering pumps; one pump is the primary dosing pump and th e second is an on-line ready spare.MEPC 60/2/9 ANNEX Page 66

I:\MEPC\60\2-9.doc Design and engineering of the BalP ure® system has considered a variety of safety measures to protect the ship’s ballasting system, the BalPure® system, as well as the health and safety of crew and technicians. Table 23 be low lists some of the primary system alarms to aid the operator in proper system function and maintenance.

Table 23 : Overview of BalPure ® System Alarms

(Table 23 is provided in the co mplete confidential dossier.)

It is important to note that because of the redundant engineering of the BalPure ® system, an alarm condition will not result in the system be ing non-operational or unsa fe. For instance, if hydrogen blower ‘A’ were to fail, an alarm would be displayed on the OIT to notify the operator, but hydrogen blower ‘B’ would automatically begin operating. This redundancy in system components ensures that the system continues to ope rate effectively and safely even if an alarm condition indicates that a system co mponent requires maintenance.

8.1.2 Chemical storage and handling

The only chemical that will require onboard stor age and handling is the neutralizing agent, sodium bisulfite. With the BalPure ® system, sodium bisulfite is st ored in stainless steel tanks designed for installation in a marine setting. Stainl ess steel is an appropriate material for storage of this chemical. Each storag e tank is properly vented with a common vent line to an alkaline ‘scrubber’, then to the atmosphere . Exact tank specifications such as size and location will be determined according to the specifics of each ship, and as approved by the ship’s classification society. The amount of bisulfite stored on boa rd will depend upon the ballasting operations of the vessel such as total ballast capacity and freque ncy, and be sized to minimize the frequency of chemical re-supply.

The only potential activities that ma y require ‘handling’ of bisulfite is either system maintenance or chemical re-supply operations. The BalPure ® system can be installed to include a flush cycle after a de-ballasti ng operation is comp leted to clear bisulfite from the chemical delivery piping, when needed to enhance the safety of the instal lation or if required by th e ship’s classification society. This greatly reduces the potential for ch emical handling during main tenance activities. Normal operation of the BalPure ® system involves automatic chemical dosing and does not require contact with any chemicals. The bisulfite fill and vent station will be located on the weather deck of the vessel; this ensures adequate ventilation. The chemi cal will be transferred only using hose(s) exclusive to bisulfite chem ical operations. Proper personal protective equipment (PPE) will be worn and only properly trained STDN technicians or representatives will be allowed to perform sodium bisulfite re-supply.MEPC 60/2/9 ANNEX Page 67

I:\MEPC\60\2-9.doc 8.1.3 Various procedures and management measures

8.1.3.1 Fire

Sodium bisulfite solution, the only chemical st ored on board, is non-flammable and poses no fire hazard. As such, it is highly unlikely that sodium bisulfite would be a direct cause of a fire on board. If a fire occurred on board in an ar ea surrounding the bisulfite storage tanks, excessive heat may result in liberation of harmful fumes (sulfur dioxide). Because the storage tanks are properly vented to the atmosphere, this does not pose any direct hazard. As a fire mitigation measure, the bisulfite storage tanks can be cool ed by applying water spray to the exterior.

In any fire situation, regardless of source or location, human e xposure to fire, smoke, fumes or combustion products will be prevented by utiliza tion of proper personal protective equipment (PPE). Also, non-essential personnel s hould be evacuated from the fire area.

During the in situ electrochemical generation of the Active Substance, hy drogen gas is created as a by-product. At concentrations >4% volume in air, hydrogen gas is fl ammable/explosive. The BalPure ® system removes hydrogen gas immedi ately and conti nuously during the electrochemical process via a degassing unit and blowers that vent to the atmosphere at a concentration of <1% vol. in air.

8.1.3.2 Accidental release measures

Technicians and ship’s crew will be sufficiently trained to res pond to a sodium bisulfite spill situation. Training will include, but may not be limited to, familiarization of the chemical properties and reactivity, exposure hazards, and correct donning of PPE. Proper PPE will include respiratory protection equipment, eye and face pr otection (goggles and/or face shield), chemical resistant gloves and boots (or boot covers ), and chemical resistant clothing.

In the unlikely event there is an accidental rel ease of sodium bisulfite, the spill should be contained and neutralized with an alkali such as sodium carbonate , sodium bicarbonate, soda ash, lime or limestone granules. All spilled material and neutralization residues must be disposed of in accordance with applicable re gulations. Decomposition of sodium bisulfite may liberate sulfur dioxide, which can be harmful when inhaled. Ade quate ventilation and respiratory protection are required in any area of accidental release.

8.1.3.3 Marine environment release

Sodium bisulfite (38% solution) is slightly acidic and should not be allowed to freely enter the marine environment as a concentrated solution. In the unlikely event of a marine environment release, sodium bisulfite is water soluble and will not cause surface impacts such as slicks or thick sludge. Spills should be reported as re quired by the approp riate agency. Storage in tanks constructed of appropriate mate rials and proper handli ng of bisulfite duri ng chemical supply operations are the primary measur es that reduce the potential risk of marine environment release.MEPC 60/2/9 ANNEX Page 68

I:\MEPC\60\2-9.doc 8.1.3.4 Conditions for controlled release

In the unlikely event that sodium bisulfite need s to be removed from the onboard storage tanks, a controlled removal utilizing the appropriate chem ical transfer pumps, discharge piping and vessel-specific safety prot ocols will be conducted.

8.1.4 Evaluation of management measures

The BalPure ® system has been properly designed and constructed for use on board ships. The risk management measures consider safe pl acement of the system on board, proper crew and technician training, and substance storage/handling that will prev ent emergency s ituations such as accidental release an d chemical exposure situations. The management measures discussed in section 8.1.3 above are considered adequate to properly manage the chemicals used with the BalPure ® system and reduce hazards to the sh ip, personnel, and the environment.

9 MATERIAL SAFETY DAT A SHEETS (G9: 4.2.7)

The hazard classifications presented in this s ection reflect reported hazards only and do not consider the actual potential for exposure to th e substances or hazards associated with the BalPure ® system. Safety mechanisms in the BalPure ® system and proper safety practices mitigate potential hazards identified. Material Da ta Safety Sheet (MSDS) and/or International Chemical Safety Card (ICSC) information is provided in appendix A.4 5

.

9.1 Active Substances hazard classification

Also known as bleach, sodium hypochlorite will be generated at a concentration of ~0.1 wt % before being injected into the ballasting system . As the sodium hypochlor ite enters the ballast system it is diluted to ~10 mg/L. Common house hold bleach is often sold in a 5% (50,000 mg/L) concentration, which is many times greater than th e concentration used with the ballast treatment system. An MSDS is not available for the chemical composition being produced inside the BalPure ® . However, ICSC #0482 (<10% solution) has b een used for hazard classification.

R-Phrases: R31, R36/38 S-Phra ses: S1/2, S28, S45, S50, S61

Sodium hypochlorite is not stor ed on board, but is generated in situ and injected directly into the ballast piping. When the system has completed a treated ballasting operation and is shut down, an automatic flush cycle clears any residual chemical from the piping. The only storage of sodium hypochlorite on board the vessel is in the ballast tanks at a conc entration of < 20 mg/L.

9.2 Hazard classification of Relevant Chemicals

9.2.1 Trihalomethanes (THMs), Ha loacetic Acids (HAAs), Bromate

THMs, HAAs and bromate disinfection by-produc ts may be formed at varying low-level concentrations in ballast water during the BalPure ® treatment process. Concentrated solutions of these substances will never be transported or uti lized on board ship. As such, typical MSDSs for these low-level substances (parts per billion range) do not accurately re present the potential

5 Appendix A.4 is provided in the confidential dossier of the proposal for approval.MEPC 60/2/9 ANNEX Page 69

I:\MEPC\60\2-9.doc hazards that may be presented by these chemical s in treated ballast water. Determining the hazards for these substances at part per billion levels is not feasible when the MSDSs reflect hazards for much higher concentrations.

Paragraph 5.3 of the Methodology seems to allo w for this justifica tion based on the above information. As such, the R-phrases and S-phra ses are not detailed fo r these disinfection by-products; however, the MSDSs are included in appendix A.4 6

.
9.2.2 Hydrogen gas

While producing sodium hypochlorite, the BalPure ® system produces hydrogen gas as a by-product. As discussed in section 8.1.1, hydrogen is removed from the sodium hypochlorite solution stream in a degassing unit as part of the BalPure ® system and vented to the atmosphere. Non-compressed hydrogen is not consider ed a hazardous material in the table in 49 CFR 172.101. A non-sparking blower in the degasser unit dilutes the hydrogen to a concentration of less than 1% (well below the 4% Lower Explosive Limit) before reaching the atmosphere. Hazard classifications a ccording to ICSC #0001 are as follows:

R-Phrases: R12 S-Phrases: S 2, S9, S16, S33 F+ symbol

9.3 Hazard classification of other chemicals

9.3.1 Sodium bisulfite

Sodium bisulfite 38% solution is added to hypoc hlorite treated ballast water as it is being discharged from the ship. This is the only subs tance stored on board. Th e quantity stored will vary depending on the amount of ba llast water treated and will be specific to each installation. Sodium bisulfite is stored in vented stainless steel tanks. Hazards according to the MSDS are as follows:

R-Phrases: R20, R22, R36, R 37, R38 S-Phrases: S26, S36

6 Appendix A.4 is provided in the confidential dossier of the proposal for approval.MEPC 60/2/9 ANNEX Page 70

I:\MEPC\60\2-9.doc 10 RISK CHARACTERIZATION AND ANALYSIS

10.1 Screening for persistence, bioaccu mulation and toxicity (G9: 5.1)

Table 24 : PBT Criteria Evaluation

Substance Persistence (P) Half-life: >60 days in marine water (MW), or >40 days in freshwater (FW) , or >180 days in marine sediment (MS),

or >120 days in freshwater sediment (FWS) (YES or NO) Bioaccumulation (B) BCF >2,000 or Log Kow ≥ 3

(YES or

NO) Toxicity (T) Chronic NOEC <0.01mg/L

(YES or NO)

Active substances

Hypochlorous acid No FW: 15 days (EPI Suite, 2009c) No Log Kow = - 0.87 No Lowest NOEC = 0.042 mg/L (Fish)

Hypobromous acid No FW: 15 days (EPI Suite, 2009b) No Log Kow = - 0.78 No based on hypochlorous acid Relevant Chemicals

Bromate No FW: 15 days (EPI Suite, 2009a) No Log Kow = - 4.6 No Lowest NOEC = 8 mg/L (Crustacean) THMs

Chloroform No FW: 4.4 days; Not expected to adsorb to sediment. (HSDB/TOXNET 2009f) No BCF = 2.9- 10.35 No Lowest NOEC = 1.5 mg/L (Fish)

Bromoform No FW: 7.1 days; Not expected to adsorb to sediment. (HSDB/TOXNET 2009l) No BCF = 14 No Lowest NOEC = 8.5 mg/L (Fish)

Dichlorobromomethane No FW: 5 days; Not expected to adsorb to sediment. (HSDB/TOXNET 2009c) No BCF = 7 No LC50 = 72 mg/L (Fish)

Chlorodibromomethane No FW: 6 days; Not expected to adsorb to sediment. (HSDB/TOXNET 2009e) No BCF = 9 No LC50 = 34 mg/L (Fish) HAAs

Bromochloroacetic acid No biodegradation data available. Not expected to adsorb to suspended solids or sediment. (HSDB/TOXNET 2009b) No Log Kow = 0.61 Data not located.MEPC 60/2/9 ANNEX Page 71

I:\MEPC\60\2-9.doc Substance Persistence (P) Half-life: >60 days in marine water (MW), or >40 days in freshwater (FW) , or >180 days in marine sediment (MS),

or >120 days in freshwater sediment (FWS) (YES or NO) Bioaccumulation (B) BCF >2,000 or Log Kow ≥ 3

(YES or

NO) Toxicity (T) Chronic NOEC <0.01mg/L

(YES or NO)

Monochloroacetic acid No FW: 14 days (Ellis, et al. 2001); Not expected to adsorb to sediment. (HSDB/TOXNET 2009d) No Log Kow = 0.22 No Lowest NOEC = 0.13 mg/L (Algae)

Monobromoacetic acid No half life data available; Not expected to adsorb to sediment. In aqueous biodegradation screening tests, reached 82% of its theoretical BOD when using pre-acclimatized inoculums (HSDB/TOXNET 2009a) No Log Kow = 0.41 No NOEC = 3.2 mg/L (Crustacean)

Dichloroacetic acid No FW: 4 days (Ellis et al. 2001); Not expected to adsorb to sediment. (HSDB/TOXNET 2009h) No Log Kow = 0.92 No EC03 = 1485 mg/L (Algae)

Dibromoacetic Acid No FW: 14 days (HSDB/TOXNET 2009g) No BCF = 0.17 No Lowest LC50 = 69 mg/L (Fish)

Trichloroacetic acid No FW: 40 days (Ellis et al 2001); Not expected to adsorb to sediment. (HSDB/TOXNET 2009m) No BCF = 0.1- 1.7 No Lowest NOEC = 100 mg/L (Algae) Other Chemicals

Sodium bisulfite No FW: 15 days (HSDB/TOXNET 2009- j) No BCF = 3.2 No Lowest LC50 = 100 mg/L (Fish)

Note: Based on an evaluation of the data presente d in Table 24 above, none of the substance meet all three criteria to be considered as PBT substances.

10.1.1 Persistence (G9: 5.1.1.1)

Using the half-life criteria of >40 days in fresh water or sediment persistence, an evaluation of the environmental persistence data for Active Substances, Relevant Chemicals, and other chemicals indicates that none of th e substances are likely to be pe rsistent in the environment. See Tables 16 and 24 above.MEPC 60/2/9 ANNEX Page 72

I:\MEPC\60\2-9.doc 10.1.2 Bioaccumulation (G9: 5.1.1.2)

Using the criteria of e ither BCF >2000 or Log Kow ≥ 3, an evaluation of the data for Active Substances, Relevant Chemicals, and other chemicals indicates that the substances have low potential for bioaccumulation. See Table 7 above.

10.1.3 Toxicity tests (G9: 5.1.2.3)

Chronic aquatic toxicity data was not located fo r hypobromous acid but toxicity is thought to be less than or similar to the chlorinated analog, hypochlorous aci d. Chronic aquatic toxicity data was not located for two THMs (bromodichloromethan e, and dibromochloromethane). Therefore, the lowest acute aquatic toxicity data points were considered. Chronic a quatic toxicity was not located for two HAAs (bromochloroacetic acid and dibromoacetic acid), and limited chronic data was located for the other HAAs. Ac ute toxicity data was also not located for bromochloroacetic acid and could not be considered. In the case of dichloroacetic acid and dibromoacetic acid, the lowest acute toxicity data point s available were considered.

With the exception of the substances noted above where chronic toxicity data was limited, all other substances did not have ch ronic NOEC values <0.01 mg/L.

11 EVALUATION OF THE TREATED BALLAST WATER (G9: 5.2)

Numerous studies have been conducted on the BalPure ® system to evaluate many aspects of treated water. In addition to the aquatic toxicity data presented in section 3, data on TRO, water quality parameters (e.g., salinity, pH, particulate organic carbon (P OC), dissolved organic carbon (DOC), total suspended solids (TSS), and disinfec tion by-products (DBPs) have been measured and recorded, and is prov ided in this section.

11.1 Total Residual Oxidants

During testing of the BalPure ® treatment system conducted at the University of Washington in 2008, varying hypochlorite concentrations were applied and TRO levels were measured at different time intervals after treatment. Table 25 below presents TRO data from three different water types that were treated. The Day 0 value represents the applied concentration of the Active Substance. As expected, the TRO c oncentration decayed quickly immediately after treatment, and then more slowly after the demand of the water was met. Regardless of water type, decay of TRO follows a predictable and consistent pattern. Also see figure 3.MEPC 60/2/9 ANNEX Page 73

I:\MEPC\60\2-9.doc Table 25 : Total Residual Oxidant (TRO) measured as Total Chlorine

Day Treatment Tank 1* TRO (mg/L) Treatment Tank 2 TRO (mg/L) Reference Freshwater 5-Dec (Day 0) 6.7 5.7 Sosik & Herwig, 2009 7-Dec (Day 2) 2.3 1.5 Sosik & Herwig, 2009 10-Dec (Day 5) 1 0.5 Sosik & Herwig, 2009 15-Dec (Day 10) 0 0 Sosik & Herwig, 2009 Brackish Water 9-Dec (Day 0) 9.1 10.5 Sosik & Herwig, 2009 11-Dec (Day 2) 6.3 N/A Sosik & Herwig, 2009 14-Dec (Day 5) 4.8 3.9 Sosik & Herwig, 2009 19-Dec (Day 10) 0 0 Sosik & Herwig, 2009 Saltwater 9-Dec (Day 0) 4.3 4.5 Sosik & Herwig, 2009 11-Dec (Day 2) 3.6 N/A Sosik & Herwig, 2009 14-Dec (Day 5) 3.1 N/A Sosik & Herwig, 2009 19-Dec (Day 10) 0 N/A Sosik & Herwig, 2009

*Samples from Tank 1 were collected for DBP and toxicity eval uation and correspond to the data presented below.MEPC 60/2/9 ANNEX Page 74

I:\MEPC\60\2-9.doc 11.2 Water quality parameters

During the University of Washington studies in 2008, water quality parameters of the treated water were measured. These included salinity, pH , POC, DOC ad TSS. In the case of fresh water and brackish water treatments, amendmen ts were made to increase the organic and suspended solids loads. Ta ble 26 summarizes the data.

Table 26 : Summary of Water Quality Parameters

Parameter Fresh water Brackish Water Seawater Reference

Initial POC (mg/L) 2.81 2.72 1.37 Sosik & Herwig, 2009 Initial DOC (mg/L) 3.72 5.69 4.23 Sosik & Herwig, 2009 Initial TSS (mg/L) 34.46 76.09 3.75 Sosik & Herwig, 2009 Salinity range (PSU) 0.62 -0.64 21.54 – 21.75 30.48 – 30.57 Sosik & Herwig, 2009

pH range 7.23 - 7.74 7.15 – 8.26 7.46 – 7.97 Sosik & Herwig, 2009 Temperature range (°C) 10.9 – 13.16 6.72 – 9.75 6.43 – 9.36 Sosik & Herwig, 2009

11.3 Disinfection By-Products (DBPs) in treated ballast water

The studies of the BalPure ® system conducted by the Univers ity of Washington in 2008 also evaluated treated water samples collected at different time intervals for THMs and HAAs. The concentrations of THMs and HAAs after treatment with the BalPure ® system of three different water types (freshwater, brackish water, and seawater) were evaluated. As stated in section 2.3.1, the presence of tribromoacetic acid was not determined analytically during pilot studies and is not anticipated to be present at en vironmentally relevant concentrations. However, tribromoacetic acid will be evaluated during land- based testing and reported in a subsequent application for Procedure (G9) Final Approval. Tables 27, 28 and 29 present data from samples collected immediately after treatment (Day 0), two days after treatment (Day 2), five days after treatment (Day 5), and ten days after treatment (Day 10). The samples collected on Day 0 and Day 2 were analysed prior to the neutra lization step. The samples collected on Day 5 and Day 10 were analysed after neut ralization with sodium bisulfite.

Bromate was also evaluated for in the brackish wate r and seawater testing cycles. The results for bromate are presented in Tables 28 and 29.MEPC 60/2/9 ANNEX Page 75

I:\MEPC\60\2-9.doc Table 27 : Levels of Trihalomethanes (THMs) and Haloacetic acids (HAAs) in Treated Fresh water

FRESHWATER
Day 0 Day 2 Day 5* Day 10* Reference
THM (EPA Method 524.2) μ g/L μ g/L μ g/L μ g/L

chloroform 21.6 42.2 52.9 43.1 Sosik & Herwig, 2009

dichlorobromomethane 33.2 70.8 90.2 73.6 Sosik & Herwig, 2009

chlorodibromomethane 71.8 127 157 123.7 Sosik & Herwig, 2009

bromoform 69.7 92.1 100 95 Sosik & Herwig, 2009

Total THM 196.3 332 400 335.4 Sosik & Herwig, 2009 HAA (EPA Method 552.1)

bromochloroacetic acid 23.4 49.7 38.3 44.2 Sosik & Herwig, 2009

monochloroacetic acid 3.9 2.5 2.7 2.8 Sosik & Herwig, 2009

dichloroacetic acid 18.4 38.8 33.7 39.5 Sosik & Herwig, 2009

trichloroacetic acid 33 42.2 47.7 56.3 Sosik & Herwig, 2009

monobromoacetic acid 2.4 3.3 3.3 2.1 Sosik & Herwig, 2009

dibromoacetic acid 36 51.1 47.4 54 Sosik & Herwig, 2009 Total HAA (6) 93.7 137.9 134.8 154.7 Sosik & Herwig, 2009

“*” = Experimental tanks we re neutralized with sodium bisulfite.MEPC 60/2/9 ANNEX Page 76

I:\MEPC\60\2-9.doc Table 28 : Levels of Trihalomethanes (THMs) and Haloacetic acids (HAAs) in Treated Brackish Water

BRACKISH WATER
Day 0 Day 2 Day 5* Day 10* Reference

THM μ g/L μ g/L μ g/L μ g/L

chloroform 0 0.5 0.6 0.5 Sosik & Herwig, 2009

dichlorobromomethane 2.4 6.4 6.3 4.6 Sosik & Herwig, 2009

chlorodibromomethane 13.4 42.8 45.7 34.1 Sosik & Herwig, 2009

bromoform 209 854 1046 782.8 Sosik & Herwig, 2009

Total THM 225 904 1099 822 Sosik & Herwig, 2009 HAA

bromochloroacetic acid 11.2 7.8 9.4 7.1 Sosik & Herwig, 2009

monochloroacetic acid 7.4 12.1 15.3 0 Sosik & Herwig, 2009

dichloroacetic acid 2.9 1.2 2.4 3.7 Sosik & Herwig, 2009

trichloroacetic acid 0 1.5 1.5 0.7 Sosik & Herwig, 2009

monobromoacetic acid 8.4 8.4 6.1 3.5 Sosik & Herwig, 2009

dibromoacetic acid 117.9 219 156.3 193 Sosik & Herwig, 2009 Total HAA (6) 136.6 242.2 181.6 200.9 Sosik & Herwig, 2009

Bromate 0 33 23 42 Sosik & Herwig, 2009

“0” = Level below detectable limits, <2 μ g/L. “*” = Experimental tanks were ne utralized with sodium bisulfite.MEPC 60/2/9 ANNEX Page 77

I:\MEPC\60\2-9.doc Table 29 : Levels of Trihalomethanes (THMs) and Haloacetic acids (HAAs) in Treated Seawater

SEAWATER
Day 0 Day 2 Day 5* Day 10* Reference

THM μ g/L μ g/L μ g/L μ g/L

chloroform 0 0 0 0 Sosik & Herwig, 2009

dichlorobromomethane 0 0 0 0 Sosik & Herwig, 2009

chlorodibromomethane 0.7 2.7 3.4 2.2 Sosik & Herwig, 2009

bromoform 32.6 106.9 117.9 90.1 Sosik & Herwig, 2009

Total THM 33.3 109.6 121.3 92.3 Sosik & Herwig, 2009 HAA

bromochloroacetic acid 0 0 0 0 Sosik & Herwig, 2009

monochloroacetic acid 5.3 12.8 9.9 0 Sosik & Herwig, 2009

dichloroacetic acid 0 0 0 0 Sosik & Herwig, 2009

trichloroacetic acid 0 0 0 0 Sosik & Herwig, 2009

monobromoacetic acid 0 1.4 2.2 0 Sosik & Herwig, 2009

dibromoacetic acid 12.5 19.6 24.7 19.3 Sosik & Herwig, 2009 Total HAA (6) 17.8 33.8 36.8 19.3 Sosik & Herwig, 2009

Bromate 0 13 23 14 Sosik & Herwig, 2009 “0” = Level below detectable limits, <2 μ g/L. “*” = Experimental tanks were ne utralized with sodium bisulfite.

The data in Tables 27 and 28 show unexpected le vels of brominated by-products, particularly bromoform, in fresh water and brackish water treatments. During these University of Washington (Sosik and Herwig, 2009) studies amendments were added to the fresh water and brackish water test runs to increase the orga nic matter and suspended solids. Amendments included were unsweetened tea, humate and test dust. These amendments were not added to the salt water test run. Although definitive analyti cal data of a potential bromide source in these amendments is not available, their addition to test waters appears to have unintentionally increased the production of disinfection by-products. Because the amendments may have been processed using unknown chemicals or may contai n other unknown contaminants, it is difficult to compare the observed disinfecti on by-product levels to those th at may occur as a result of treating waters with naturally present organic matter.MEPC 60/2/9 ANNEX Page 78

I:\MEPC\60\2-9.doc For the brackish water test run, where higher leve ls of bromoform were measured, there is an additional factor likely contributing to the levels of bromin ated organic by-produc ts observed. To reach the desired brackish water salinity, natura l seawater was diluted with tap water from a municipal water source. This water had been previously chlorinated using chlorine gas. The researchers aerated the water to remove pre-existing chlorine; however, at the pH of the test water most of the residual chlorine pres ent in the tap water would exist as HOCl/OCl - and some HOBr/OBr - if the chlorine had a small bromin e impurity. Therefor e, use of local chlorinated tap water potentially increased the level of hypochlorit e ion in treated water and may also be a source of bromine. Additionally, the tap water source lik ely had pre-existing levels of disinfection by-products. Disinfection by-product data for th e specific date the tap water was used by the researchers is not known, but the an nual report from the mu nicipal water treatment facility indicates that leve ls of THM’s and HAA’s detect ed during 2008 were 23.2 μg/L and 15.5 μg/L, respectively, (City of Port Townsend, 2008).

Due to the addition of amendments , the potential presence of bromine present in the chlorine source prior to treatment with the STDN sy stem, along with the presence of pre-existing disinfection by-products, it can be reasonably deduced that these fact ors influenced the levels of brominated by-products in the fresh water and brackish water studies. Further analysis of disinfection by-products will be cond ucted during land-based testing.

11.4 Toxicity testing of treated ballast water
11.4.1 University of Washington studies

Toxicity testing of water treated with the BalPure ® system has been conducted by Nautilus Environmental (Nautilus, 2009). In conjunction with BalPure ® system mesocosm studies undertaken at the University of Washi ngton (Sosik and Herwig, 2009), Nautilus evaluated treated water collected at differing time interval s after treatment and after neutralization with bisulfite. The mesocosm studies were done w ith three different water types: fresh water, brackish, and seawater. Acute aquatic toxicity te sting was performed with algae, invertebrates, and fish for water samples collected immediatel y after treatment with hypochlorite (Day 0), 2 days after hypochlorite tr eatment, and 5 days afte r hypochlorite treatment. These samples were collected and tested prior to the neutralization step (i.e. pre-neutraliza tion). After the Day 5 samples were collected, residual oxidants in the me socosms were neutralized with bisulfite and a second Day 5 sample was collected for testing. Fi nally, 5 days after neut ralization (i.e. 10 days after hypochlorite treatment) additional samples were collected for toxicity testing. The toxicity data is presented in section 3. Water from th e mesocosm studies was also evaluated for TRO, water quality parameters, and disinfection by-p roducts (DBPs). The data are presented in Tables 25, 26, 27, 28 and 29.

As expected, toxicity was observ ed for all species in the samp les collected immediately after hypochlorite treatm ent (Day 0). Some toxicity was observed in the Day 2 and Day 5 pre-neutralization samples, with the exception of fr eshwater fish. With th e exception of one data point for the marine diatom ( Skeletonema ) in brackish water, toxicity was not observed in the Day 5 post-neutralization sample s. Although the sensit ivity between the taxo nomic groups (algae, invertebrate, and fish) varied with the water type (fresh water, brackish, and se awater) tested, fish were the most tolerant sp ecies and invertebrates were generally the least tolera nt species. Overall, the most sensitive species tested was Daphnia magna (Nautilus, 2009). In all sp ecies tested, no toxicity was observed in the sample s tested 5 days after neutralization (Day 5+).MEPC 60/2/9 ANNEX Page 79

I:\MEPC\60\2-9.doc Because sodium bisulfite is injected into the ba llast water discharge line at a slight excess to ensure that all residual oxidants are neutraliz ed, Nautilus Environmental performed toxicity testing using a very large excess of bisulfite (Nautilus, 2005). Seawater samples treated with sodium hypochlorite generated with the BalPure ® system were provided to Nautilus by the University of Washington on behalf of STDN. Test solutions were pr epared by diluting the chlorinated sample with laboratory seawater to a total chlorine concentra tion of 3 mg/L, and then neutralizing with a 22 mg/L solution of sodium bisulfite. Although only 6 mg/L of bisulfite would be needed to neutralize a 3 mg/L hypochlorit e treatment, the substant ially higher bisulfite addition was evaluated to determine a safe ty margin for the ne utralization process (Nautilus, 2005). A 0.5X dilution series was us ed to obtain a definitive toxicity threshold. Toxicity tests were conducted using 7-day chronic (sub-lethal endpoints) tests with a mysid ( Mysidopsis bahia ) and a fish ( Menidia beryllina ), a 48-hour larval de velopment test using a mussel ( Mytilus edulis) , a 48-hour germination and germ tube length test using giant kelp ( Macrocystis pyrifera) , and a 16-day embryo development test using Pacific herring ( Clupea pallasi ). Results were that none of the test endpoints exhibi ted a 25% reduction relative to the control. These data suggest that a 3 mg /L treatment of hypochlorit e in seawater with up to 22 mg/L of sodium bisulfite would not be expected to exhib it an adverse effect, defined as resulting in a 25% performance reduction for the e ndpoint tested. Based on this study, the safety factor = 22 /application rate (m g/L sodium bisulfite) was determ ined. This safety factor is considered conservative because no effects were observed at this treatment level (Nautilus, 2005).

Evaluating toxicity to aquatic organisms at various holding times after treatment (without neutralization) demonstrates that the residual chlori ne remains effective after treatment. Additionally, performing toxicity testing pre- and post- neutralization verifies that the bisulfite treatment is effectively removing residual oxidants making treated water discha rge environmentally safe. Lastly, dosing with excessive neutralizing agent, as well as holding neutralized water for an additional 5 days and then performing toxicity testing, demonstrates that the neutralizing agent is not itself toxic or otherwise making the neutralized discharge toxic to aquatic organisms.

Documentation of the quality assurance program fo r toxicity testing at Nautilus Environmental is included in appendix A.7 7

. 11.4.2 Naval Research Laboratory studies

Chronic toxicity tests were conducted by Nautilu s Environmental (Nautilus, 2007) using treated ballast water samples collected during the full-s cale study at the Naval Research Facility in Key West, Florida (United States). Samples were co llected from challenge te sts in which organism and organic enriched ballast water was treated with hypochlorite to a 15 mg/L total residual oxidant (TRO) concentration and then held for 48 hours (2 days). After 48 hours the treated water was neutralized with an excess concentration of ~ 2 mg/L sodium bisulfite. Five concentrations of treated sample using a 0.5X dilution series were tested. Bioassays were conducted using 7-day tests with a mysid ( Americamysis bahia ) and a larval fish ( Menidia beryllina ), a 48-hour larval development test using a mussel ( Mytilus galloprovincialis ), and a 48-hour germination and germ tube le ngth test using the giant kelp ( Macrocystis pyrifera ). Chronic toxicity tests using mysid shrimp and silverside minnows were conducted according to US EPA (2002) methods. Chronic toxicity te sts using mussels and giant kelp were conducted according to US EPA (1995).

7 Appendix A.7 is provided in the confidential dossier of the proposal for approval.MEPC 60/2/9 ANNEX Page 80

I:\MEPC\60\2-9.doc Results for the chronic toxicity tests are summari zed in Table 5. No toxicity was observed with the treated water samples for any species teste d. The highest concentration with no observed effect (NOEC) was 100 percent sample (Nautilus, 2007).

11.5 Determination of holding time

Because the BalPure ® system employs an oxidant neutraliza tion step at discharge, there is no specified holding time required before treated water can be safely discharged to the environment. Using ORP technology (discussed in section 8.1.1) sodium bisulfite can be added to the ballast water discharge line at the concentration need ed to ensure that all residual oxidants are neutralized. Aquatic toxicity testing perf ormed with treated water samples pre- and post-neutralization with sodium bisulfite demonstr ates that the discharge has no adverse effect after the neutralization process. Toxicity tes ting results with excessive treatment of sodium bisulfite also indicate that there is no adverse eff ect from sodium bisulfite. The data is presented in Tables 3 and 5.

Therefore, the holding time that tr eated ballast water needs to be retained is determined by the amount of contact time required to properly disinfect and inactive AIS. Studies performed at the University of Washington determined that at low (<2 mg/L) TOC concentrations 98%-100% of organisms were inactivated 5 hours after hypochlorite tr eatments of 2.95 mg/L–3.71 mg/L (Herwig et al. , 2006). Studies performed at the Naval Research Laboratory determined BalPure ® hypochlorite treatments of 14 mg /L–16 mg/L effectively inactivat ed >99% of organisms 24 hours after treatment in seawat er with 10 mg/L TOC (NRL, 2007). Thes e studies indicate that even if a ship had to discharge treated ballast water shor tly after hypochlorite treat ment, the number of live organisms would be very limited and no aquatic t oxicity could be expected from neutralized discharge.

11.6 Reaction with organic matter

As discussed in section 2, chlorine-based water disinfection can re sult in the formation of THMs and HAAs. The concentration of these DBPs that will result from di sinfection is based on the Active Substance dose as well as the parameters of the water being treated (e.g., TOC, pH, temperature), with the amount of TOC being of significa nt influence. Lower TOC levels reduces the amount of DBPs formed. The BalPure ® system filters incoming ballast water, which will reduce the amount of TOC, and thereby, the DBPs formed duri ng disinfection. The levels of THMs and HAAs in treated ballast water at va rious doses of Active Substance in different water types and water quality conditions is pr esented in section 11.3. The concentrations of THMs and HAAs in treated water discharge will be monitored further during land-based testing at NIOZ. Results of this testing will be re ported in the future dossier for Final Approval.

11.7 Characterization of degradat ion route and rate (G9: 5.3.5)

For all substances related to use of the BalPure ® system, both the biotic and abiotic degradation data are shown in Table 16. For THMs and HAAs, biotic degradation for all of these simple halogenated organic compounds is on the order of da ys to weeks. Abiotic degradation of these compounds is longer. In the ca se of bromate and sodium bisu lfite, no specific degradation information was located in the existing literature. However, for these inorganic substances, the BCF, Koc, and Log Kow values indicate low pot ential for bioaccumulation, low adsorption to sediment, and high water solubility.MEPC 60/2/9 ANNEX Page 81

I:\MEPC\60\2-9.doc 11.8 Prediction of discharge and environmental concentrations

Due to the oxidant neutralization step during ballast water discharge, the TRO in treated water discharge will be non-detectable. Therefore, the discharge and predicted environmental concentration (PEC) for TRO is 0 mg/L.

Sodium bisulfite is added to balla st water discharge at a slight ex cess to ensure that any residual oxidants have been reacted. As such, treated water discharge may contain sodium bisulfite. In seawater bisulfite is redu ced to bisulfate, and ultimately to sulfate ion as shown in Reactions 14-16 in section 2.4. In the marine environment, sulfate ion is ubiquitous and does not pose unacceptable environmental risks. Therefore, discharge or environmental concentrations are not considered relevant.

During the pilot studies of the BalPure ® system, the amounts of DBPs in treated water discharge were evaluated in three different water types (fresh water, brackish water and seawater), at different Active Substance treatments and organi c matter levels. The data is presented in sections 11.1 to 11.3 above. In fresh wate r, total THMs in tr eated water ranged from 196.3 μg/L – 400 μg/L and total HAAs ra nged from 93.7μg/L–154.7 μg/L. In brackish water, total THMs ranged from 225 μg/L–1099 μg/L and total HAAs ranged from 136.6 μg/L–242.2 μg/L. In seawater, to tal THMs ranged from 33.3 – 121.3 μg/L and total HAAs ranged from 17.8 – 36.8 μg/L. The DBP bromate was also measured in the brackish and seawater studies. In brackish water, brom ate concentrations range d from <2 – 42 μg/L. In seawater, bromate concentrations ranged from <2 μg/L–23 μg/L.

As discussed in section 5.4, formation of DBPs is dependent upon several variables such as Active Substance dose, contact time, pH, or ganic matter, etc. Data from the BalPure ® discharge reflects those trends. For instance, the highest THM and HAA formation occurred in the brackish water treatment where the hypochlorit e dose, organic matter, and suspended solids were the highest. Similarly, the lowest THM a nd HAA formation occurred in the seawater study where the hypochlorite dose, organic matter and suspended solids were at lower levels. Additional contributing f actors to the levels of brominated by-products observed are discussed above in section 11.3.

The DBP concentrations from the pilot studies noted above consider analysis of the whole effluent (i.e. discharge concentration) only and do not reflect actual enviro nmental concentrations when treated ballast water is released in a harbour where di lution and mixing will occur. Meaningful PEC values will be derived using a ppropriate emission scenario modelling based on analysis of treated ballast water samples drawn from the full-scale land-based test setup of the-BalPure ® system and will be presented in a future dossier for Final Approval.

Regardless of the Relevant Chemical discharge c oncentrations, it is important to note that acute and chronic (sub-lethal endpoint) toxicity testing of the neut ralized whole effluent from the BalPure ® system was not toxic to aquatic organisms, with the only exception being Skeletonema (EC50 66.1%, Table 3) in one fresh water study.MEPC 60/2/9 ANNEX Page 82

I:\MEPC\60\2-9.doc 11.9 Assessment of potential for bioaccumulation

Based on the data shown in Table 7 for both Activ e Substances and Relevant Chemicals, all of these substances have very low BCFs, indicati ng that they are unlikely to bioaccumulate in aquatic organisms or present a significant food chai n risk. With the exception of the THMs, all of the Active Substances and Relevant Chemicals have very low log Kow’s, which indicates the compounds are hydrophilic. The THMs, as woul d be expected, are m oderately hydrophobic. However, none of the log Kow values estimat ed and/or located in the literature are ≥ 3.

11.10 Effects assessment

The data for the substances associated with use of the BalPure ® system indicate that there is a low potential for bioaccumulation, sediment ad sorption, and persistence in the aquatic environment. No effects or risks in the form of secondary (food chain) poisoning or to sediment species are anticipated. As such, aquatic toxic ity presents the only likely potential risk for aquatic organisms.

Whole effluent toxicity testing (acute and sub- lethal endpoints) of ne utralized ballast water discharge suggests no apparent toxicity for a ll species and endpoints tested (E(L)C50 and/or NOEC values of ≥ 100% ballast water sample) with the exception of one marine diatom, Skeletonema costatum (96-hour exposure EC50 of 66.1% sample) (Nautilus, 2009). In light of these results, although aquatic toxi city is identified as having the most potential for risk to aquatic organisms, toxicity testing results suggest the potential risk is low.

11.11 Effects on aquatic organisms

A thorough literature review of available acute and chronic toxi city data for the substances associated with the STDN system was conducted; the data are presented in Tables 2 and 4. However, the data set is limited for many of the substances. Even where data was located in literature studies did not meet va lidity criteria, it could only be used as supporting information. This resulted in incomplete data sets for fis h, crustaceans, and algae that prevented individual PNEC derivation for all substances based on th e guidance in the Methodo logy (MEPC 58/2/7) as well as the Technical Guidance Document on Risk Assessment (2004). Similar data gaps are also noted in other dossiers s ubmitted for Procedure (G9) a pproval, suggesting that other literature searches have had similar findings.

When a complete valid data set was located in literature, and/or any supporting data could be identified, the reported toxicity values have been used for PNEC derivation. Where the available data is limited, the PNEC derivation approach used in the EU RAR for Sodium Hypochlorite (2007) was utilized.

Two valid acute aquatic toxicity data points (o ne fish, one crustacean) were located for TRO (saltwater), with one additional acute data point for alga e used as supporting information. Additionally, one valid chronic fish test was lo cated, while one chronic endpoint for crustacean and one chronic endpoint for algae were located as supporting information. The lowest acute effect concentration reported was 0.062 mg/L. Using a higher assessment factor (1000) because the data is based on acute endpoints and the alga e exposure period was only 24 hours, results in a PNEC for TRO (saltwater) of 0.062 μg/L. This corresponds directly with the PNEC saltwater value derived in the sodium hypochlorite RAR; also 0.06 μg/L (as TRC).MEPC 60/2/9 ANNEX Page 83

I:\MEPC\60\2-9.doc For hypochlorous acid (HOCl), a complete data set of acute toxicity end points was located. One valid chronic end point for fish was also locate d, and one sub-lethal data point for a crustacean was available as supporting information. Us ing the lowest acute effect concentration (0.032 mg/L) and an assessment factor of 1000, a PNEC value of 0.032 μg/L was derived. No data for hypobromous acid was located in the literature; however, the aquatic toxicity is expected to be similar or le ss than that of the chlorinated analogue, hypochlorous acid. Therefore, a PNEC value of 0.032 μg/L hypobromous acid is proposed.

For bromate, acute aquatic toxicity endpoints we re located for the three basic taxonomic groups, and two chronic endpoints were located (one fish, one alga). Although not all of the data met validity criteria, a conservative PNEC of 13.6 μg/L was derived using the lowest acute endpoint (13.6 mg/L) and an assessment factor of 1000. Us ing the available data and a conservative assessment factor allows for PNEC derivation and avoids performing unnecessary toxicity testing, particularly for vertebrate species.

For THM’s, the data sets for dibromochlor omethane (DBCM) and dichlorobromomethane (DCBM) were incomplete, while chloroform and br omoform had complete data sets that would allow for direct PNEC calculation. Complete, valid acute and chronic toxicity data sets for fish, crustacean, and algae were available for chlo roform. Using the lowest chronic effect concentration of 1.5 mg/L and an assessment f actor of 100 results in a chloroform PNEC of 15 μg/L. A complete, valid acute toxicity data set was av ailable for bromoform, and two chronic endpoints (one fish, one alga) were available as s upporting information. Using the lowest acute effect concentra tion (12.3 mg/L) and an assessmen t factor of 1000, results in a bromoform PNEC of 12.3 μg/L.

As mentioned above, the data sets for DBCM a nd DCBM are incomplete. Only one valid acute endpoint for fish was located for both DBCM and DCBM. It seems beyond the scope of Procedure (G9) application process to conduct chem ical-specific testing with these substances on a variety of organisms to complete the data set av ailable in open literature. This is particularly true considering that whole efflue nt toxicity testing results of th e treated ballast water discharge are available. Therefore, as an alternative to having PNEC values th at are non-derivable for DBCM and DCBM, the PNEC derivation approach for THMs used in the EU RAR for sodium hypochlorite was utilized. For chloroform, the RAR references an aquatic PNEC of 146 μg/L, which is higher than that calculated from the chronic end point data located during the literature search (chloroform PNEC 15 μg/L). However, the RAR presents that the data collected in connection with an assessment for seawater chlo rination “...suggest that the ecotoxicities of the brominated THMs are not markedly different from chloroform”. (EU RAR for sodium hypochlorite, 2007). The RAR concludes that for the purposes of a broad assessment, THMs can be regarded as having similar toxicities and applies the chloroform PNEC to total trihalomethanes. Therefore, utilizing th e RAR suggested PNEC value of 146 μg/L for total THMs ensures that all THMs are considered, even in cases where toxicity data is limited. To maintain a conservative approach, the lower PNEC values for chloroform (15 μg/L) and bromoform (12.3 μg/L) derived from the availa ble data can be used for PEC/PNEC ratio evaluation for these two THMs using the chemical analysis data collected during land-based testing. This evaluation will be presented in a fu ture dossier for Procedure (G9) Final Approval.

With the exception of one HAA, trichloroacetic ac id (TCA), the aquatic toxicity database is incomplete and did not allow for direct PNEC calculation for each individual HAA. For TCA, the literature search located two valid acute end points (one fish, one crustacean), and one acute endpoint for an algae that was reported as a NOEC rather th an an EC50. Additionally, two chronic endpoints were located for algae as supporting information. Us ing the lowest acuteMEPC 60/2/9 ANNEX Page 84

I:\MEPC\60\2-9.doc effect concentration of 500 mg/L and an assess ment factor of 1000, a PN EC of 500 μg/L was calculated for TCA.

Again, it is beyond the scope of Procedure (G9) application process to conduct the extensive amount of toxicity testing that would be required to complete the HAA data set. This is particularly true considering that whole effluent toxicity testing results of the treated ballast water discharge are available. Theref ore, the approach utilized in the sodium hypochlorite RAR for PNEC derivation of HAA’s was util ized. That is, the RAR considered it “...conservative to treat all haloacetic acids simplistica lly as being TCA...” in regards to PNECs and all HAA’s could be considered to have a PNEC of 0.85 μg/L. Si nce this PNEC is far lower than the PNEC of 500 μg/L based on the data fo und in literature, this seems to be a very protective and reasonable approach to this assessment.

For sodium bisulfite, the availa ble aquatic toxicity data was al so limited. Two acute endpoints for fish were located along with three acute endpoi nts for crustaceans. The lowest acute effect occurred at 100 mg/L. No chroni c aquatic toxicity data was loca ted. Therefore, a PNEC value appears to be non-derivable. However, base d on the chemical properties (high solubility, low BCF value, low Koc value), and also consider ing that <2 mg/L of s odium bisulfite may be present in treated ballast water discharge, it is highly unlikely that this substance will cause unacceptable environmental risks during use of th e STDN system. This has been demonstrated in whole effluent toxicity testing treated wi th sodium bisulfite (see Tables 3 and 5 and section 11.4.1). Should further information or tes ting results become available to derive a PNEC for sodium bisulfite, this will be reflected in a subsequent application for Procedure (G9) Final Approval.

Table 30 : PNEC Value Summary

Substance Lowest Effect Concentration (mg/L) Assessment Factor PNEC Value ( μ g/L) Reference/Comments Active Substances

TRO (Freshwater) Insufficient Data Located -- 0.06 (as TRC) Based on EU Sodium Hypochlorite RAR suggested PNEC for TRC in freshwater TRO (Saltwater) 0.062 1000 0.062 HOCl 0.032 1000 0.032

HOBr Data Not Located -- 0.032 Estimated based on HOCl toxicity data Relevant Chemicals Bromate 13.6 1000 13.6 THMs

Chloroform 1.5 100 15 / 146 146 μg/L to be used as PNEC for Total THMs

Bromoform 12.3 1000 12.3 / 146 146 μg/L to be used as PNEC for Total THMs

Dibromochlorometh ane (DBCM) Data Not Located -- Not Derivable Use EU Sodium Hypochlorite RAR suggested PNEC for Total THMsMEPC 60/2/9 ANNEX Page 85

I:\MEPC\60\2-9.doc Substance Lowest Effect Concentration (mg/L) Assessment Factor PNEC Value ( μ g/L) Reference/Comments

Dichlorobromometh ane (DCBM) Data Not Located -- Not Derivable Use EU Sodium Hypochlorite RAR suggested PNEC for Total THMs

Total THMs -- -- 146 Based on EU Sodium Hypochlorite RAR suggested PNEC for Total THMs HAAs

Bromochloroacetic acid Data Not Located -- 0.85 Based on EU Sodium Hypochlorite RAR suggested PNEC for HAAs

Monochloroacetic acid Insufficient Data Located -- 0.85 Based on EU Sodium Hypochlorite RAR suggested PNEC for HAAs

Dichloroacetic acid Insufficient Data Located -- 0.85 Based on EU Sodium Hypochlorite RAR suggested PNEC for HAAs Trichloroacetic acid 500 1000 500

Monobromoacetic acid Insufficient Data Located -- 0.85 Based on EU Sodium Hypochlorite RAR suggested PNEC for HAAs

Dibromoacetic acid Insufficient Data Located -- 0.85 Based on EU Sodium Hypochlorite RAR suggested PNEC for HAAs Other Chemicals

Sodium bisulfite Insufficient Data Located -- Not Derivable

11.12 Effects on sediment

Based on the chemical and physical properties of the substances associated with the BalPure ® system, no effects on sediment are an ticipated. See sections 3.4 and 5.2.

11.13 Comparison of effect assessm ent with discharge toxicity

The information reviewed for degradation and bi oaccumulation of the substances related to use of the BalPure ® system suggest that potential eff ects from these mechanisms cannot be reasonably anticipated. The effects assessment esta blishes that there is pot ential for reaction of the Active Substances with organic matter and form ation of DBPs. This has been confirmed by data from ballast water discha rge samples analysed for DBPs. The presence of DBPs in disinfected water can potentially result in toxic e ffects to aquatic organisms. With the exception of testing with one species ( Skeletonema ), all toxicity tests after neutralization with bisulfiteMEPC 60/2/9 ANNEX Page 86

I:\MEPC\60\2-9.doc resulted in E(L)C50 and/or NOEC values ≥ 100% ballast water sample (see data in Tables 3 and 5, discussion in section 11.4). Tes ting using a 96-hour exposure with Skeletonema resulted in an EC50 of 66.1% sample. Thus, to reach a co ncentration lower than the EC50 would require less than a two-fold diluti on. This level of diluti on will be readily achieve d when treated water is discharged.

Discharge toxicity testing results are consistent with the overall effects assessment that the potential effects are related to aquatic toxicity, rather than bioaccumulation or persistence in the environment. Further, the toxicity observed wi th neutralized discharge samples was limited to only one of the nine species tested.

12 RISK ASSESSMENT

The risks presented by having the BalPure ® Ballast Water Management System on board are discussed in section 12.1 and the risks to huma n health presented in section 12.2. Section 12.3 addresses risks to the aquatic environment.

12.1 Risk to Safety of Ship

A risk analysis was conducted by The Glosten Associates and approved by American Bureau of Shipping (ABS) to demonstrate that the BalPure ® Ballast Water Management System is acceptable for installation on tank ers, and specifically on the S/R American Progress

. A qualitative comparative analysis was conducted for the chemical dosing and ballast water sampling systems involved in the installation. Re sults indicated that these systems do not pose greater risk than comparative systems curre ntly allowed by the ABS Steel Vessel Rules (Glosten Associates, Inc., 2006).

12.1.1 Corrosion

Corrosion of steel and other metalli c parts is of significant concern to the shipping industry. This is why a majority of the surfaces in ternal to the ship and in contact with seawater such as piping and tanks are coated with protective paints an d coatings. Tests of how well these coatings protect metal surfaces are completed by various organizations and coating vendors themselves. One study prepared test samples from standard sh ip building steels with fully intact coatings applied to manufacturer’s speci fications (BMT 2004). Four diffe rent coatings and solutions containing 0, 10 mg/L, and 40 mg/L Total Residual Chlorine (TRC, as added hypochlorite) were tested to determine weight gain as a measure of permeability. All samples showed the same rate and minimal amount of water uptak e, indicating the coated parts were not permeable and did not corrode when in contact with solutions cont aining up to 40 mg/L TRC. Other tests were conducted applying ASTM Stan dard “Test Method for Evalua tion of Painted or Coated Specimens Subjected to corrosive Environments D 1654-92”. F our different coating systems using five identical test coupons per coating we re exposed to two levels of hypochlorite. Scribed and unscribed test coupons were subjected to four differe nt environments to replicate conditions in a ballast tank bei ng: (1) buried in in ert sand to replicate the under sediment condition at the bottom of the silt filled tanks; (2) in damp humid space replicating an empty ballast tank; (3) fully submerged to replicate a full ballast tank; and 4) rotating on a wheel with cyclic immersion into the test solution to replicat e the splash zone in the top of a partially full tank. Results show: (1) no accelerated failures in any unscribed coating system; (2) other samples exposed to hypochlorite tend to experien ce slightly more damage than the control samples; and (3) there is a trend in the level of damage realized relative to the tank location with more aggressive locations provides more damage. But a statistical an alysis of the data shows thatMEPC 60/2/9 ANNEX Page 87

I:\MEPC\60\2-9.doc at the 95% confidence level only the 40 mg/L e xposure on the wheel showed significantly more damage (BMT 2004).

There also have been studies with uncoated steel to determine the effects of fresh water and saltwater with and without the presence of chlori ne. One report contains corrosion experiments again simulating the four conditions observed in a ballast tank: submerged in sediment, submerged in liquid, exposed to damp humid space above the liquid, and alternating between submerged liquid and humid air. TRC levels in the liquids were maintained at 10 mg/L and 40 mg/L and the tests were conducted for up to 90 days (BMT 2004). Results indicate that the condition had more of an impact on the corro sion rate than the addition of hypochlorite. As an example, the corrosion rate submerged in seawater was approximately 0.75 mm/year over 90 days. However, another sample tested in a cyclic manner showed corrosion rates of 2.8 mm/year over 90 days. Similar samples e xposed to 10 mg/L TRC (an expected maximum instantaneous threshold for the BalPure ® system in a ballast tank) showed corrosion rates of 0.80 mm/year and 3.25 mm/year, respectively.

Paik developed a time dependent corrosion wastage model for tankers and Offshore Floating Production and Storage Vessels (FPSO’s) in whic h corrosion loss is calculated from the following equation:

t r

= C 1 (T – T c – T t ) C 2

where the thickness of the steel t r at time T after construction is dependent on the time to coating breakdown T c and the transition time T t and coefficients C 1 and C 2 (Paik 2004, as referenced in BMT 2004). Paik investigated 34 different components within the typical ship structure and completed a statistical review of ships up to 32 y ears old. The study determined that the time to coating breakdown and corrosion is generally 7.5 years after constr uction. The rate of corrosion is linear, however this is depe ndent on location within the tank. After applying the corrosion rates given in the test above to the Paik model it was concluded that hypochlorite in levels commensurate with ballast water treatment will re duce the time for steel replacement on a ship by less than 5% (Paik 2004, as referenced in BMT 2004).

12.1.2 Fire and explosion

Potential fire and explosion risks associated with the BalPure ® system are related to the hydrogen gas generated during the electroche mical process, the sodium sulfite (neutralizing agent) stored on board or electrical com ponents of the system.

While producing sodium hypochlorite, the BalPure ® produces hydrogen gas as a by-product. Hydrogen gas can be explosive at concentrations >4% volume in air. The hydrogen is removed from the sodium hypochlorite solution stream in a degassing unit as part of the BalPure ® system. Non-compressed hydrogen is not consider ed a hazardous material in the table in 49 CFR 172.101. A non-sparking blower in th e degasser unit dilutes the hydrogen to a concentration of less than 1% (well below the 4% Lower Explosive Limit) before reaching the atmosphere.

The BalPure ® system has several safety mechanisms that are engineered into the system to ensure that hydrogen gas is effectivel y removed from the ship. Thes e include a patented cyclone separator device that removes the hydrogen gas fr om the biocide solution (see section 8.1.1) and redundant blowers to reduce the co ncentration of hydrogen gas to <1 % within a vent line to the atmosphere outside the ship. Additionally, a hydrogen gas detect or is located in the space(s)MEPC 60/2/9 ANNEX Page 88

I:\MEPC\60\2-9.doc Exposure Assessment

Review & Assessment of Toxicological Data

Risk Quantification Uncertainties Evaluation Risk Assessment Conclusions Hazard Identification (Data Review/ COPC Selection ) where hydrogen gas may enter should a BalPure ® system failure occur. The PLC monitors and controls all of these safety features and will result in an alarm condi tion should the system require maintenance.

Sodium bisulfite is a non-flamma ble, stable liquid. Excessive he ating and/or cont act with acids can liberate sulfur dioxide fumes, bu t does not create a fire hazard.

All electrical components and hard ware associated with a BalPure ® unit have been tested and certified to meet established safety guidelines by groups such as UL, IEC, IEEE, and FM. These same components have been approved for use a board ship by classification societies such as ABS, USCG, Lloyd’s, and BV.

With the inclusion of the above safety measures , the fire/explosion risk associated with having the BalPure ® system installed on board is low.

12.1.3 Storage of chemicals

As presented in section 2, the only chemical stored on board is sodium bi sulfite. As discussed above, sodium bisulfite is non-flammable and does not present any onboard fire risk. Because the BalPure ® system is engineered and constructed to ensure proper chemical storage, and will be installed per approval of the ship’s classificati on society, the potential ri sks to the ship from chemical storage are very minimal.

12.2 Risks to human health

12.2.1 Introduction

Quantitative human health risk assessments ge nerally follow the process outlined below:

Step 1 Step 2 Step 4 Step 5 Step 6

Step 3

With respect to the potential human hea lth risks associated with the BalPure ® BWMS, Step 1 is the hazard identification process. The BalPure ® operational system is reviewed, and chemicals stored on the ship and added to the system or produced in situ at concentrations potentially harmful to human health are carri ed forward in the quantitative risk assessment as the chemicals of potential concern (COPCs). In Step 2 , COPC-specific toxicity values for use in the quantitative risk analysis are co mpiled via an exhaustive literatu re search. If no toxicity data exists, information from appropriate surrogates and/or structurally similar chemicals may be used. For COPCs without toxicity information, quantitative risk evaluation is not possible.MEPC 60/2/9 ANNEX Page 89

I:\MEPC\60\2-9.doc In Step 3 , exposure scenarios are developed: (1) to describe the potentia l exposures on board during routine vessel operations including ope ration, monitoring and maintenance of the BalPure ® BWMS; and (2) to describe the potential exposures on and off board during equipment failure/and or accidental release of the COPCs; and (3) to provide a basis for quantifying those exposures. Each exposure scenario addresses th e COPCs, the potential route or mechanism of exposure, and potentially exposed human populations (known as “receptors”). When operation-specific data for scenario deve lopment are unavailable, conservative values found in the appropriate regul atory guidance are used.

In Step 4 , the toxicity and exposure assessments are in tegrated into quantita tive expressions of risks. This includes COPC-specific, multi-pathway risks for each of the potential receptors. The risk values presented in a risk assessment are conditional estimates derived from a considerable number of conserva tive, health-protective assumptions about exposure and toxicity. Thus, to place the risk estimates in proper perspective, it is impor tant to specify the assumptions and uncertainties inherent in the risk assessment. Th is process is conducted in Step 5

. This step may also involve the re-evaluation of data or the identification of additional data requirements to decrease uncertainty. Step 6 involves the development and pres entation of conclusions that can be inferred from the findings of the risk assessmen t. This step provides risk managers with insight into the interpretation of the risk assessment results.

12.2.2 Hazard identification/chemica l of potential concern selection

The chemicals of potential concern (COPCs) associated with the use of BalPure ® BWMS are selected by determining which chemicals, either stored and added to the system or formed in situ
, have the potential to adversely impact human health under either routine operating conditions, maintenance operations, or during an emerge ncy resulting in an unexpected release.

As described in sections 1 and 2, the BalPure ® system is a nearly closed system that produces the disinfection agent, sodium hypochlorite (as a mi xture of hypochlorous ac id/hypobromous acid), via electrolysis of seaw ater. Hydrogen gas, a secondary by-product of the in situ electrochemical process, is vented to the atmosphere (using blower s) at concentrations of less than 1%. However, exposure to atmospheric gases typically are not included in risk assessment, and the safety features inherent in the BalPure ® system as well as the physical properties of hydrogen gas suggest it is unlikely to accumulate to levels of concern (e.g., above the LEL). Therefore, as indicated in section 2.3.3, hydrogen gas is not considered relevant for the assessment of human health risks. Sodium bisulfite is added to de-chlorinate the ballast water as it is discharged from the ship. As identified in section 2, the fo llowing chemicals associated with the BalPure ® system were evaluated as possible COPCs, and the rational for their select ion or exclusion as COPCs is provided.

The final list of COPCs carried forward in th is risk assessment includes hypochlorous acid, THMs, HAAs, bromate and sodium bisulfite.

12.2.2.1 Chemicals produced i n situ on vessel

.1 Active Substance : Sodium hypochlorite (N aOCl) is generated at a concentration of ~0.1 wt % (1,000 mg/L) prior to inje ction into the ballast tanks. As sodium hypochlorite enters th e ballast system, it is diluted to no more than 20 mg/L (See section 2.2.1 fo r discussion of determination of applied hypochlorite concentration) . Because sodium hypochlorite isMEPC 60/2/9 ANNEX Page 90

I:\MEPC\60\2-9.doc generated in situ , it is not stored on board the vess el except in the ballast tanks at concentrations <20 mg/L. At the pH of seawater, and because seawater contains bromide ion, the actual disi nfecting agents are hypochlorous acid (HOCl) and hypobromous acid (HOBr). These two chemicals, along with chloramines and bromamines, are referre d to as the total residual oxidant (TRO) and are evaluated in this ri sk assessment as a single COPC – hypochlorous acid.

.2 Trihalomethanes and haloacetic acids : Once mixed with ballast water, sodium hypochlorite can react with orga nic materials to form trihalomethanes (THMs) and haloacetic acids (HAAs). BalPure ® -associated concentrations of these chemical groups have been genera ted for treatment of fresh-, brackish, and seawater (Sosik and Herwig, 2009). THMs detected in clude chloroform, bromoform, bromodichloromethane, and chlorodibromomethane, at a maximum total THM concentration of 1.1 mg/L (in brackish water following neutralization on Day 5). Six HAAs we re detected, at a maximum total HAA concentration of 0.24 mg/L (in brackish water pre-ne utralization on Day 2). Refer to section 11.3, Tables 27-29.

. 3 Bromate : Bromate has been measur ed as a DBP in BalPure ® treatment of brackish water and seawater (Sosik and Herwig, 2009). The maximum concentration, 0.042 mg/L, occurred in brackish water test ed on Day 10. Refer to section 11.3, Tables 28-29.

12.2.2.2 Chemicals stored on board

.1 Sodium Bisulfite : Sodium bisulfite (NaHSO 3 ), as a 38% aque ous solution, is loaded aboard from a filling station and stored in holding tanks which are vented to the atmosphere. The sodium bisulfite is injected into the ballast line during de-ballasting operat ions to neutralize any re sidual TRO. Sodium bisulfite is added in excess (<2 mg/L) to ensure complete neutralization of the TRO. Given that sodium bisulfite is stored on the ship and residual remains in the ballast water, it is incl uded as a COPC in the risk assessment.

12.2.3 Human Exposure Scenario

The US EPA (1989) defines an exposure pathway as “the course a chemi cal or pollutant takes from the source to the organism exposed”. An ex posure route is “the way a chemical or pollutant enters an organism after contact” (US EPA 1989 ). A complete exposure pathway requires four key elements: chemical sources; migration rout es (i.e. environmental transport); potentially exposed human receptors; and routes of exposure to impacted media (e.g., ingestion of chemicals in water). All four factors are required for a complete exposure pathway; if any one factor is missing, the pathway is considered incomplete. Because an incomplete pathway does not pose a potential health hazard, incomplete exposure path ways are not included in this health risk assessment. Potential human receptor groups and exposure routes are described below.

12.2.3.1 Potential exposure pathways

The conceptual exposure model (C EM) provides the basis for a co mprehensive evaluation of the risks to human health by iden tifying the mechanisms through wh ich people may be exposed to COPCs. The CEM traces the COPCs in a logical migration from their sources through variousMEPC 60/2/9 ANNEX Page 91

I:\MEPC\60\2-9.doc release mechanisms and exposure routes to po tentially affected receptor groups. The CEM identifies the exposure routes that are potentially complete under the given use(s). These pathways are evaluated in the quantitative risk assessment for each receptor. The CEM also facilitates the analysis and screening of exposure pathways not likely to po se significant risks.

Primary, secondary, and tertiary sources of COPCs associated with the BalPure ® system are listed in the CEM (figure 7). The prim ary sources of COPCs include the in situ production of sodium hypochlorite, the sodium bisulfite storage tanks and associated pi pelines, and the treated ballast water itself. Chemicals associated with the BalPure ® system are expected to remain contained, unless a storage or pipeline failure occurs. Su ch a release would allow COPCs to migrate to secondary and tertiary sources. For example, accidental spills and pipe or tank failure could result in the release of COPCs on board the vessel or to seawater . From these secondary sources, the COPCs may migrate to tertiary sources such as volatizing into air within enclosed spaces on the vessel or in outdoor air, or migrati on to shorelines and/or docking areas.

As shown in figure 7, only exposure to COPCs resu lting from leaks of st orage tanks, pipelines and ballast tank discharges are eval uated quantitatively in this risk assessment. This is because no exposure is anticipated during normal operating conditions, because chemicals used in the BalPure ® system are contained within a closed system (hypochlorous acid is produced in situ and is injected directly into the ballast system; s odium bisulfite is injected into the ballast water as it is discharged from the ship), While both the chemical storage tanks and ballast tanks are vented, vent lines are directed to the atmosphe re or to spaces within the ship with proper ventilation. Significant volatiliz ation of COPCs from solution (f rom either storage tanks or treated ballast water) into indoor or outdoor air is extremely unlikely, and is not evaluated quantitatively. It is noted, how ever, that the trihalomethanes produced as disinfectant byproducts are volatile and hydrophobic, and as such, coul d be inhaled near ballast water or vent air outlets. While this inhalation pathway is not evaluated in the quantitative risk assessment, potential exposures and risks associated with vent air during ballastin g and de-ballasting are discussed in section 12.2.5.5.

SOURCES

Primary Secondary Tertiary

Vessel Surface

Seawater

Dock/Shoreline



### • Pathway is or might be complete; sufficient data available for quantitative evaluation

X Pathway is or might be complete, but judged to be minor; not evaluated quantitatively

1 STDN Technician exposure considered worst-case scenario for anyone working onboard vessel.

2 Not evaluated quantiatively in risk assessment; semi-qualitatively evaluation provided.

Pathway is not complete; no evaluation required General Public Off Board

Dock Workers



### • X Spills/Leaks from Storage Tanks, Pipelines or BWTS Vapors/Indoor Air

Vapors/Outdoor Air



### • STDN Technician

X 1

X



### • Release from Ballast Tanks

Dermal Contact/ Incidental Ingestion X Inhalation X On Board

X Dermal Contact/ Incidental Ingestion

Inhalation

X Exposure Routes



### •

X 2 Ship's Crew

Figure 7 : Conceptual Exposure Model, BalPure ® SystemMEPC 60/2/9 ANNEX Page 92

I:\MEPC\60\2-9.doc 12.2.3.2 Identification of potentially exposed populations

As described in sections 1.1 and 8.1.1, the BalPure ® system is designed to operate automatically, with the system processes during ballasting and de-ballasting started and stopped based on electronic signals. Therefor e, under normal operating condi tions, neither STDN BalPure ® technicians nor the ship’s crew is expected to have contact with the system-related COPCs. Because the BalPure ® system is closed system, exposure to the Active Substance, disinfection by-products, or sodium bisulfite is highly unlikely.

As shown in figure 7, people with the grea test potential for exposure to BalPure ® BWMS-related COPCs are STDN BalPure ® technicians involved in routine main tenance and repair, or chemical resupply activities. Although unlik ely, ship’s crew on board and in the spaces where the system components are present could also be exposed to COPCs during a pipeline or chemical tank failure; it is anticipated that these potential exposures would be less than those for STDN technicians conducting routine maintenance or ch emical resupply. Because ballast water tanks remain closed at all times (with the exception of planned inspections/repairs of empty tanks), STDN technicians and the ship’s crew are not expect ed to have direct cont act with treated ballast water. However, either the ship’s crew or dock workers could have dermal contact with released ballast water as a result of spray drift. As shown in figure 7, while STDN technicians, ship’s crew, or dock workers potentially could inhale chemicals volatilizin g from solution, these source-exposure pathways are cons idered highly unlikely and are not evaluated qua ntitatively.

Others with potential for COPC contact include the general public recreating at beaches near areas where treated ballast wate r has been discharged from th e ship, although this exposure scenario is highly unlikely.

These receptors and the potential expos ure pathways are summarized below:

.1 STDN BalPure ® system technicians: STDN technicians or properly trained BalPure ® personnel are the only receptors who would work directly with the system, including routine maintenanc e and chemical resupply of sodium bisulfite, and therefore ha ve the greatest potential for actual exposure. Although proper chemical storage and handling, safety training, a nd use of appropriate PPE would prevent direct cont act with COPCs, technici ans could be exposed to hypochlorous acid, sodium bisulfite, or tr eated ballast water during maintenance activities via:

- incidental ingestion of COPCs duri ng routine maintena nce and chemical resupply activities;
- dermal contact with COPCs during routine maintenance and chemical resupply activities; and
- inhalation of COPC vapors or mists in air during routin e maintenance and chemical resupply activities.

The potentially complete and quantifia ble exposure pathways for STDN BalPure ® technicians include: (1) incidental in gestion of and dermal contact with hypochlorous acid at maximum concentrati ons of 1,000 mg/L, and (2) incidental ingestion of and dermal contact with sodi um bisulfite during chemical resupply at maximum concentrations of 380,000 mg/L.MEPC 60/2/9 ANNEX Page 93

I:\MEPC\60\2-9.doc For hypochlorous acid, potential exposure is further mitigated by a pipeline flushing cycle that is engineered into the system. For instance, after generation of the Active Substances and the ballasting operation is complete, a 10-minute cycle is initiated by the PLC to the electroly zer cells and injection pipelines with seawater. The same can be done for the sodium bisulfite delivery piping when a de-ballasting operation is complete, if needed to enhance the safety of an installation. These flush cycles ensure that pipelines do not remain filled with chemicals when the system is not in operation.

.2 Ship’s crew : Under normal operating conditions, the ship’s crew has limited to no interaction with the BalPure ® system. The system and piping are located in areas where the crew may be present, but normal ballasting and de-ballasting modes of the BalPure ® system are controlled automatically and do not require chemical contact. However, this group has the potential to be exposed to sodium bisulfite while chemical-resupp ly activities take place, or if a pipeline or storage tank fails, or to any COPCs dur ing a failure of the BalPure ® system. Exposure to treated ballast water is highly unlikely, since the tanks are opened for required inspections only when empty, although the potential for exposure to ballast water spray drift exists. The actual exposure potential for this group is extremely low, but the possible exposur e routes include:

- incidental ingestion of the COPC s during an accidental release;
- dermal contact with COPCs during an accidental release; and
- inhalation of COPC vapors or mists in air during an accidental release.

As previously indicated, accidental re lease of hypochlorous acid and/or sodium bisulfite are expected to result in expo sures smaller than those for STDN BalPure ® technicians engaged in routine maintenan ce and chemical resupply. Moreover, as described in section 8.1.3.2, STDN technicians and ship’s crew will be trained to respond to sodium bisulfite spills; therefore these exposure scenarios are not included in the quantitative risk assessm ent. The only potentially complete and quantifiable exposure pathway for ship’s cr ew is dermal contact with COPCs in discharged ballast water, including hypochlorous acid, bromate, THMs, HAAs, and sodium bisulfite at the maximum re sidual ballast water concentrations – <20, 0.042, 1.1, 0.242, and <2.0 mg/L, respectivel y (see section 11.3). It is also important to note that treated ballast wa ter discharge will be neutralized with sodium bisulfite, so the maximum resi dual ballast water concentration for hypochlorous acid is overly conservati ve because it does not consider neutralization.

.3 Dock workers : Under normal operating conditi ons, dock workers will have no interaction with the BalPure ® system or chemical resupply activities. While exposure to treated ballast water is hi ghly unlikely, the potential for exposure to ballast water spray drift exists. The act ual exposure potential for this group is extremely low, but the possible exposure routes include:

- dermal contact with COPCs in ballast water spray drift.MEPC 60/2/9 ANNEX Page 94

I:\MEPC\60\2-9.doc The only potentially complete and qua ntifiable exposure pathway for dock workers is dermal contact with COPCs in discharged ballast water, including hypochlorous acid, bromate, THMs, HAAs, and sodium bi sulfite at the maximum residual ballast water concentrations. This quantitative evaluation is identical to that for ship’s crew.

.4 General public : Although it is unlikely that th e general public would recreate near docked ships, it is possible that r ecreation areas could be located downstream of shipping docks. While any COPC releas es from the ship would be immediately diluted into surrounding seawat er, the general public (adu lts or children) could be exposed to COPCs via:

- incidental ingestion of ballast water COPCs discharged to seawater near the shoreline;

- dermal contact with ballast water CO PCs discharged to seawater near the shoreline; and
- inhalation of ballast water COPC vapor s or mists in outdoor air near the shoreline.

The potentially complete and quantifia ble exposure pathways for swimmers include: incidental inges tion of and dermal contact with hypochlorous acid, bromate, THMs, HAAs, sodium bisulfite, and re cently discharged ballast water. It is conservatively assumed that maximum residual ballast water COPC concentrations are diluted 100-fold prior to contact with beachgoers.

Table 31 provides a summary of th e exposure point concentrations discussed above and used in the risk assessment.

Table 31 : Summary of COPC Exposure Point Concentrations

STDN Technicians:

Maximum Concentration in System Ship’s Crew/Dock Workers: Maximum Residual Ballast Water Concentrations General Public: Diluted Residual Ballast Water Concentration COPC mg/L mg/L mg/L Hypochlorous Acid 1.00E+03 2.00E+01 2.00E-01 Bromate 4.20E-02 4.20E-04 THMs 1.10E+00 1.10E-02 HAAs 2.42E-01 2.42E-03 Sodium bisulfite 3.80E+05 2.00E+00 2.00E-02

12.2.3.3 Quantitative exposure assessment

As described above, although highly unlikely, STDN BalPure ® technicians could have direct contact (ingestion and dermal contact) with COPCs within the system during maintenance activities and to sodium bisulf ite during chemical resupply activities. Ship’s crew or dock workers could have dermal exposure to ballast wa ter spray drift, and beac hgoers may recreate inMEPC 60/2/9 ANNEX Page 95

I:\MEPC\60\2-9.doc the vicinity of ballast water di scharges. Intake factors are used to determine the amount of COPC that each receptor is potentially exposed to via each exposure route, and are used to evaluate both cancer risk and non-cancer hazard.

The following equation was used to quantify in take from the dermal contact pathway:

I

w = (C w )(SA)(PC)(CF)(ET)(EF)(ED) / (BW)(AT)

Where I w = intake from dermal contact with COPC in ballast water/solution (mg/kg-d) C w = maximum concentration of a CO PC in ballast wate r/solution (mg/L) SA = skin surface area in cont act with ballast water/solution (cm 2 /d) PC = dermal permeability coefficient (cm/hr), chemical-specific CF = conversion factor 10 -3 (L/cm 3 ) ET = exposure time (h/d) EF = event frequency (d/y) ED = exposure duration (y) BW = body weight (kg) AT = averaging time (d), ED x 365d/y (noncarcinogens), 70y x 365d/y (carcinogens)

The following equation was used to quantify inta ke from the incidental ingestion pathway:

I

w = (C w )(IngR)(EF)(ED) / (BW)(AT)

Where I w = intake from incidental ingestion of COPC in ballast water/solution (mg/kg-d) C w = maximum concentration of COPC in ballast water/solution (mg/L) IngR = ingestion rate (L/day) EF = event frequency (d/y) ED = exposure duration (y) BW = body weight (kg) AT = averaging time (d) - ED x 365d/y (noncarcinogens), 70y x 365d/y (carcinogens)

The receptor-specific exposure parameters used to estimate COPC intake based on the equations above are summarized below and the resulting inta ke factors summarized in Tables 32 and 33. As shown in Tables 32 and 33, the intake equa tions described above for the general public are age-adjusted to account for six year s of exposure as a child and 24 y ears of exposure as an adult. This approach is consistent with guidance (US EPA 1989 and 1991e), and accounts for any differences in exposure assumptions between children and adults.

.1 STDN technicians

The adult exposure factors used to qua ntify potential exposur e to hypochlorous acid during maintena nce of the BalPure ® system or to sodium bisulfite during chemical resupply are:MEPC 60/2/9 ANNEX Page 96

I:\MEPC\60\2-9.doc - Ingestion rate: While incidental ingestion is likely to be only a fraction of the ingestion rate for swimming, 0.05 L/hour is conservative and therefore used (US EPA 1989);

- Skin surface area available for contact : During an accidental spill, technicians or workers may have signi ficant contact with hands and arms, which results in a skin surface area of 3,300 cm 2 (US EPA 2004);

- Permeability constant : Chemical-specific; for inorganics without a permeability coefficient, and especially for ionized chemicals, guidance recommends using 0.001 cm/hour (US EPA 2004);

- Body weight : The adult body weight typicall y applied in risk assessment is 70 kg (US EPA 2002);

- Exposure time : The exposure time is assumed to be 0.25 hours per event, is based on best professional judgment;

- Event frequency : Based on best professional judgment, an exposure frequency of 1 spill event per year is used;

- Exposure duration: Based best professional ju dgment, the exposure duration is 10 years; and

- Averaging time: The life expectancy of 70 years (25,550 days) was used as the averaging time for exposure to carc inogenic contaminants (US EPA 2002). The averaging time for non-carcinogenic effects is equal to the exposure duration of 10 years (3,650 days).

.2 Ship’s crew/dock workers

The adult exposure factors used to quan tify potential exposure to discharged ballast water spray drift are:

- Skin surface area available for contact : During a spray drift incident, crew or workers may have limited contact with hands, arms, and face, which results in a skin surface area of 1,800 cm 2 which is based on best professional judgment (10% of total body; US EPA 2004).

- Permeability constant : Chemical-specific; for inorganics without a permeability coefficient, and especially for ionized chemicals, guidance recommends using 0.001 cm/hour (US EPA 2004). The value for chloroform, 0.0068 cm/hour, is used as a surrogate for THMs and HAAs (US EPA 2004).

- Body weight : The adult body weight typically applied in risk assessment 70 kg (US EPA 2002);

- Exposure time : The exposure time is assumed to be 0.25 hours per event, is based on best professional judgment;MEPC 60/2/9 ANNEX Page 97

I:\MEPC\60\2-9.doc - Event frequency : Based on best professional judgment, an exposure frequency of 1 spray drift event per year is used;

- Exposure duration : Based on best professi onal judgment, the exposure duration is 10 years.

- Averaging time : The life expectancy of 70 years (25,550 days) was used as the averaging time for exposure to carc inogenic contaminants (US EPA 2002). The averaging time for non-carcinogenic effects is equal to the exposure duration of 10 years (3,650 days).

.3 General public (beachgoers)

The exposure factors used to quantify poten tial exposure to CO PCs in discharged ballast water to swimming b eachgoers (adults and children) are described below. As discussed above and shown in Tables 32 and 33, the intake factors used to evaluate cancer risk for beachgoers are age- adjusted to account for differences in exposure assumptions between children and adults.

- Ingestion rate : The ingestion rate for swimming, 0.05 L/hour, is used (US EPA 1989).

- Skin surface area available for contact: The average of the 50th percentile total skin surface area for adults is 18,000 cm 2 and for children is 6,600 cm 2 (US EPA 2004).

- Permeability constant: Chemical-specific; for i norganics (hypochlorous acid and sodium bisulfite) without a permeab ility coefficient, and especially for ionized chemicals, guidance r ecommends using 0.001 cm/hour (US EPA 2004). The largest value for organics evaluated in this risk assessment is for chloroform, at 0.0068 cm/hour (US EPA 2004).

- Body weight: The adult and child body weight s typically applied in risk assessment are 70 and 15 kg, respectively (US EPA 2002).

- Exposure time: The US EPA’s recommended swimming duration of 60 minutes per event, based on the 50th percentile value (US EPA 1997).

- Event frequency: Based on best professional judgment, an upper-end exposure frequency of 10 events per year is used.

- Exposure duration: Based on US EPA guidance, the exposure duration is 30 years, which includes six years as a child and 24 years as an adult (US EPA 2002).

- Averaging time: The life expectancy of 70 years (25,550 days) was used as the averaging time for exposure to carc inogenic contaminants (US EPA 2002). The averaging time for non-carcinogenic effects is equal to the exposure duration; 24 years as an adult (8,760 days ) and 6 years as a child (2,190 days).MEPC 60/2/9 ANNEX Page 98

I:\MEPC\60\2-9.doc Table 32 : Summary of Dermal Intake Factors

IF dermal =

IF dermal/adj =

IF dermal = Dermal Intake Factor, L water/kg body weight-day SA = Surface Area, cm 2 PC = Dermal Permeability Constant, cm/hour CF = Volumetric Conversion Factor, 1 Liter/1000 cm 3 ET = Exposure Time, hours/day EF = Exposure Frequency, days/year ED = Exposure Duration, years BW = Body Weight, kg AT = Averaging Time, days

Child A dult (0-6 years)

SA 3300 1800 18000 6600
CF 0.001 0.001 0.001 0.001
ET 0.25 0.25 1 1
EF 1 1 10 10
ED 10 10 24 6
BW 70 70 70 15
AT

carcino g ens 25550 25550 25550 25550 AT

nonarcinogens 3650 3650 8760 2190

PATHWAY-SPECIFIC INTAKE FACTORS: Chemical-Specific Intake Factors via Dermal Contact (IF dermal ), L water/kg body weight-day
Carcinogens x PC 4.61E-06 2.52E-06 3.45E-03 NA Noncarcinogens x PC 3.23E-05 1.76E-05 7.05E-03 1.21E-02 Exposure Variable General Public STDN Technician Ship's Crew/ Dock Worker SA x PC x CF x ET x EF x ED BW x AT

SA

child x PC x CF x ET child x EF child x Ed child BW child x AT + SA dult x PC x CF x ET adult x EF adult x ED adult BW adult x ATMEPC 60/2/9 ANNEX Page 99

I:\MEPC\60\2-9.doc Table 33 : Summary of Oral Intake Factors

IF oral =

IF oral/adj =

IF oral = Oral Intake Factor, L water/kg body weight-day IR = Ingestion Rate, L/hour ET = Exposure Time, hours/day EF = Exposure Frequency, days/year ED = Exposure Duration, years BW = Body Weight, kg AT = Averaging Time, days

Child Adult (0-6 years) IngR 0.05 NA 0.05 0.05
ET 0.25 NA 1 1
EF 1 NA 10 10
ED 10 NA 24 6
BW 70 NA 70 15
AT carcino g ens 25550 NA 25550 25550 AT nonarcinogens 3650 NA 8760 2190

PATHWAY-SPECIFIC INTAKE FACTORS: Chemical-Specific Intake Factors via Oral Ingestion (IF oral ), L water/kg body weight-day
Carcinogens 6.99E-08 NA 1.45E-05 NA Noncarcinogens 4.89E-07 NA 1.96E-05 9.13E-05 Exposure Variable General Public
STDN Technician Ship's Crew/ Dock Worker IngR x ET x EF x ED BW x AT

IngR child x ET child x EF child x ED child BW child x AT + IngR adult x ET adult x EF adult x ED ad u BW adult x AT

12.2.4 Health effects in humans

The objective of this section is to provide info rmation regarding the potential for health risks from exposure to chemicals potentially present in treated ballast water. Specifically, this section describes how dose-response, or toxicity values , are established and used for non-carcinogenic and carcinogenic COPCs. Because only the inges tion and dermal exposure routes are evaluated quantitatively, this section focuse s exclusively on oral toxicity va lues (which are commonly used as surrogates for evaluation of dermal exposure).

12.2.4.1 Acute health effects

All of the COPCs identified can be skin and eye irritants as described in Table 9. With the exception of sodium bisulfite, all of the COPCs identified are generated in situ at aqueous concentrations in the μg/L to mg/L range. These in situ generated COPCs are at concentrations too low to cause serious eye and skin irritation. There may be some lim ited potential for minor eye irritation if there is direct contact with ba llast water, due to a combination of the COPCs, salts, and other compounds which may be in th e ballast/seawater prior to treatment. Skin irritation is considered unlikely given the very low concentrations of the COPCs. THMs, with chloroform an example of a solv ent known to de-fat th e skin, are known to cause both skin andMEPC 60/2/9 ANNEX Page 100

I:\MEPC\60\2-9.doc eye irritation when present at much hi gh concentrations than those present in situ in ballast water (IPCS 2004).

The one COP, which could represent an acute ey e and skin hazard, is the sodium bisulfite solution stored on the vessel. The MSDS (Tab le 9) clearly states that contact with the 38% solution can cause eye and skin irrita tion. Appropriate PPE should be used when working with this solution to minimize the poten tial for these acute health effects, and only trained personnel should be handling the c oncentrated solution (Mallinckrodt 2006a).

The potential for acute irritation of the nose an d throat upon inhalation is unlikely. The THMs are the most likely COPCs to volatize and are cons idered in a separate inhalation assessment in Table 39. The safe levels used in the inha lation assessment (section 12.2.5.5) are considered protective of irritation of mucous membranes caused by volatile THMs (OSHA 2009).

The more critical endpoint for potential exposure to ballast water COPCs at low concentrations are chronic health issues such as liver disease and cancer. These chronic effects are fully addressed in the risk assessment.

12.2.4.2 Non-carcinogenic adverse health effects

For the non-carcinogenic effects of specific constituents, most regu latory agencies assume a dose exists below, which no adverse health eff ects will be seen (US EPA 1989). Below this “threshold” it is believed that exposure to a chem ical can be tolerated w ithout adverse effects. Adverse effects manifest only when physiolo gic protective mechanisms are overcome by exposure to doses above the threshold. The re ference dose (RfD), e xpressed in units of milligrams per kilogram-day (mg/kg-d), repres ents the daily intake of a constituent (averaged over a year) per kilogr am of body weight that is belo w the effect threshold for the constituent. It is assumed that non-carcinoge nic exposure doses are not cumulative from age group to age group over a lifetime of exposure (US EPA 1989). An RfD is specific to the constituent, route of exposure, and duration over which the exposure occurs.

Agencies, most notably the US EPA, review a ll relevant human and animal studies for each constituent and select the studies pertinent to th e derivation of specific RfDs. Each study is evaluated to determine the no-observable-adv erse-effect level (NOA EL) or, if data are inadequate for such a determin ation, the lowest-observable-a dverse-effect level (LOAEL). The NOAEL corresponds to the dose (mg/kg-d) that can be administered over a lifetime without inducing observable adverse effects. The LOAEL corresponds to the lowest daily dose (mg/kg-d) that can be administered over a lifetime that induces an observable adverse effect. The toxic effect characterized by the LOAEL is referred to as the “cr itical effect” (US EPA 1989).

To derive an RfD, the NOAEL (or LOAEL) is divided by uncertainty factors (alternatively known as safety factors) to ensure protection of human health. Uncertainty factors are applied to account for: (1) extrapolation of data from laboratory animals to humans (interspecies extrapolation); (2) variation in human sensitivity to the toxic effects of a constituent (intraspecies differences); (3) derivation of a chronic RfD ba sed on a subchronic rather than a chronic study; and (4) derivation of an RfD from the LOAEL rather than the NOAEL. A safe ty factor of 10 is typically applied for each of these uncertainties during the development of the RfD. Thus, the safety factor for an individual COPC could be as high as 10,000. In additi on to these uncertainty factors, modifying factors betw een 0 and 10 may be applied to reflect additional qualitative considerations (US EPA 1989).MEPC 60/2/9 ANNEX Page 101

I:\MEPC\60\2-9.doc 12.2.4.3 Carcinogenic adverse health effects

The incremental lifetime can cer risk (ILCR) attributed to a ca rcinogen is calculated as a product of the daily intake (mg/kg-d) and the cancer slope factor (CSF). The US EPA’s model of carcinogenesis assumes the relationship between exposure to a carcinogen and cancer risk is linear over the entire dose range, except at very high doses (US EPA 1989). This linearity assumes there is no threshold-of-exposure dose be low which harmful effects will not occur. Because of this, carcinogenic effects are considered to be cumulative across age groups when considering lifetime exposures.

CSFs are upper-bound (95% upper confidence limit (UCL)) estimates of the increased cancer risk per unit dose, in which risk is expressed as the probability that an individual will develop cancer within his or her lifetime as the result of exposure to a given level of a carcinogen. All cancers or tumors are considered whether or not death results. This approach is inherently conservative because of the no-threshold assumption and the use of the 95% UCL of the estimated slope of dose versus cancer risk.

To identify oral RfDs and slope factors, the studies and appro aches described by the US EPA in its Integrated Risk Information System (IRIS) we re compared to the toxicological information reported in section 4 (Tables 11 through 13). When no toxicity values are re ported in IRIS, best professional judgment was used to develop valu es based on the data reported in Tables 11 through 13.

12.2.4.4 Hypochlorous acid and hypobromous acid

The health effects asso ciated with hypochlorite (salts of hypochlorous aci ds) have been studied primarily as it relates to its use as a disinfectant in drinking wate r. While various drinking water goals for disinfectants have been established by the US EPA, the World Health Organization (WHO), and the European Union (EEC), only the US EPA has identified an oral reference dose for chlorine: 0.1 mg/kg-day (US EPA 1994). This value, as reported in IRIS, is based on the NOAEL of 14 mg/kg-day for rats exposed to ch lorine in drinking water for two years (NTP 1992a), as reported in Table 11 (Chroni c Mammalian Toxicity). The US EPA (1994) applies a composite unc ertainty factor of 100: 10 to account for inte rspecies extrapolation and 10 to account for the protection of sensitive subpopulations.

No toxicological information was found for hypobromous acid, or the associated hypobromite salts. As previously reported, hypobromous acid is believed to be similar in toxicity to its chlorinated analogue, hypochlorous acid. Theref ore, as indicated in the COPC selection section 12.2.2, the Active S ubstance in the BalPure ® system is evaluated qualitatively as hypochlorous acid.

12.2.4.5 Bromate

The US EPA has identified an oral refere nce dose for bromate in IRIS: 0.004 mg/kg-day (US EPA 2001a). This value is based on th e NOAEL of 1.1 mg/kg-day for rats exposed to potassium bromate in drinking water for 100 w eeks (US EPA 2001a), as reported in Table 11 (Chronic Mammalian Toxicity). The oral referen ce dose includes a composite uncertainty factor of 300: 10 to account for extrapolation from animal data to humans, 10 to protect sensitive subpopulations and account for potential differenc es between adults and children, and 3 to account for some deficiencies in the database (US EPA 2001a).MEPC 60/2/9 ANNEX Page 102

I:\MEPC\60\2-9.doc As summarized in Table 13 (Summary of Data on Carcinogenicity), IARC has concluded that bromate is a possible human carci nogen, while the US EPA classifies it as a probable human carcinogen. Based on the same study of rats e xposed to potassium bromate in drinking water for 100 weeks (and also summarized in Table 13), the US EPA (2001a) includes an oral slope factor of 0.7 per mg/kg-day in IRIS, based on the incidence of renal tubular tumors, thyroid follicular tumors, and testicular mesotheliomas.

12.2.4.6 Total Trihalomethanes

As discussed in section 2, trihalomethane (T HM) by-products in ballast water may include chloroform (trichloromethane), bromoform (t ribromomethane), dichlorobromomethane, and chlorodibromomethane. The US EPA has identifie d oral reference doses for all four compounds in IRIS with values ranging from 0.0 1 mg/kg-day (chloroform; US EPA 2001b) to 0.02 mg/kg-day (bromoform [US EPA 1991a], dichlorobromomethane [US EPA 1991b], and chlorodibromomethane [US EPA 1991c ]). The lowest (and therefor e most conserva tive) value is from the study of beagles administered oral doses (15 or 30 mg/kg-day) of chloroform in capsules for 5 days/week over 7.5 years for which a LOAEL of 15 mg/kg/day fo r liver effects, as is reported in Table 11 (Chronic Mammalian Toxicity). 8 The US EPA (2001b) derived the oral reference dose based on a modeled low benchmark dose (BMDL 10 ) of 1.0 mg/kg-day, and included a composite uncertainty factor of 100: 10 to account for extrapolation from animal data to humans and 10 to account for intr ahuman variability (US EPA 2009). 9

As reported in Table 13 (Summary of Data on Ca rcinogenicity) either IARC or the US EPA has classified each of the four THMs as “probable” or “possible” human car cinogens. Based on the studies identified in Table 13, US EPA, in IRIS, has derived or al slope factors for bromoform (7.3 x 10 -3 per mg/kg-day; US EPA 1991a), dichlorobromomethane (6.2 x 10 -2 per mg/kg-day; US EPA 1991b), and chlorodibromomethane (8.4 x 10 -2 per mg/kg-day; 1991c) 10 . The highest value is the most conservative, therefore the oral slope factor for chlorodibromomethane is used to evaluate carcinogenic risks from THMs.

12.2.4.7 Total Haloacetic Acids

As described in section 2, halo acetic acid (HAA) by-products in ballast water may include bromochloracetic acid, mono-, di- a nd trichloroacetic acids, and m ono- and dibromoacetic acids. Of these, the US EPA (2003) has identified an oral reference dose in IRIS for only dichloroacetic acid, however, this va lue is based on a subchronic (90 da y study), which is not optimal for the evaluation of chronic effects. As described abov e, a chronic effects level can be developed based on the NOAELs or LOAELs reported for chroni c toxicity studies in Table 11. The NOAELs identified for HAAs in Table 11 range from 3.5 mg/kg/day for monochl oroacetic acid (EU 2005) to 40 mg/kg/day for trichloroacetic acid (IPCS 2000a ); all studies are based on effects on rodents, therefore a composite uncertainty factor of 100 ( 10 to account for extrapolation from animal data to humans and 10 to account for intrahuman variabil ity) is appropriate for the determination of a reference dose from any of th ese studies. The lowest (and therefore most conservative) NOAEL-based calculated value is for monochlor oacetic acid – 0.034 mg/kg/day. The LOAELs

8 IRIS reference doses for bromoform and chlorodibromomethane are based on subchronic studies associated with the same NTP studies (1989 and 1985, respectively) cited in Table 11. The IRIS reference dose for dichlorobromomethane is based on the NTP (1987) chronic LOAEL of 25 mg/kg/d cited in Table 11.

9 A similar oral reference dose resulted from the NOAEL/LOAEL approach.

10 US EPA has determined that while or al exposure to chloroform may cause cancer, the reference dose of 0.01 mg/kg-day is considered to be protective against cancer risk.MEPC 60/2/9 ANNEX Page 103

I:\MEPC\60\2-9.doc identified for HAAs in Table 11 range from 2 mg/kg/day for dibromoacetic acid (NTP 2007) to 40 mg/kg/day for dichloroacetic acid (IPCS 2000a ); all studies are base d on effects on rodents, therefore, a composite uncertainty factor of 1000 (10 to account for the use of a LOAEL instead of a NOAEL, 10 to account for extrapolation from animal data to humans, and 10 to account for intrahuman variability) is appropriate for the dete rmination of a reference dose from any of these studies. The lowest (and therefore most conser vative) LOAEL-based value is for dibromoacetic acid – 0.002 mg/kg/day. Because the LOAEL-based va lue results in the lowest reference dose, it is used to evaluate non-carci nogenic risks from HAAs.

As reported in Table 13 (Summary of Data on Carcinogenicity) only dichloroacetic acid, dibromoacetic acid, and trichloroacetic acid have been classified as either “probable” or “possible” human carcinogens. 11 However, IRIS includes oral slope factors for only dichloroacetic acid (0.05 per mg/ kg-day; US EPA 2003), therefore this value is used to evaluate carcinogenic risks from HAAs. It is noted that while no cancer sl ope factor has been developed for the remaining HAAs, the effects levels reported in Table 13 are up to 20 times lower for dibromoacetic acid ( ≥ 4 mg/kg/day) than those reported for dichloroacetic acid (84 mg/kg/day). Therefore, some uncertainty exists in the carcinogenic evaluation of HAAs; specifically carcinogenic risks associated with dibromoacetic acid, the dominant residual HAA measured in ballast water (see Tables 27 to 29) may be underestimated.

12.2.4.8 Sodium bisulfite

The US EPA has not developed an oral reference dose for sodium bisulfite inclusion in IRIS. However, as reported in Table 11 (Chronic Mammalian Toxicity), The Health Council of the Netherlands identified a NOAEL of 72 mg/k g-day for rats fed up to 20,000 mg/L sodium bisulfite, based on local effect s (blood in feces; hyperplastic ch anges of gastric epithelium) (2005). In the absence of other information, an oral RfD can be calculated by applying a composite uncertainty factor of 100, 10 to account for interspecies extrapolation and 10 to account for intrahuman variability, to the NOAEL ( 72 mg/kg-day). This results in an oral RfD of 0.72 mg/kg-day.

As reported in Table 13 (Summary of Data on Carcinogenicity), sodi um bisulfite is not classifiable as to its carcinogeni city, therefore, no oral cancer slope factor is available.

As shown in Table 34, reference doses were identif ied for all COPCs. Ca ncer slope factors are available for only bromate, THMs, and HAAs; ther efore, carcinogenic effects can be evaluated for only these three COPCs.

11 As indicated in Table 13, NTP (2009) reports “Cl ear evidence of carcinogeni c activity” in animals for bromochloroacetic acid.MEPC 60/2/9 ANNEX Page 104

I:\MEPC\60\2-9.doc Table 34 : Summary of Toxicity Criteria

Chemical Oral Reference Dose (mg/kg/day) Oral Cancer Slope Factor (per mg/kg-day) Hypochlorous Acid 1.00E-01 (chlorine) NC

Bromate 4.00E-03 7.00E-01 THMs 1.00E-02 (chloroform) 8.40E -02 (chlorodibromomethane) HAAs 2.00E-03 (dibromoacetic acid) 5.00E-02 (dichloroacetic acid) Sodium bisulfite 7.20E-01 NC NC – No criteria

12.2.5 Risk characterization

Risk characterization, the final step in the risk assessment process, combines data from the hazard identification, health effects assessmen t, and exposure assessment to estimate the potential carcinogenic and non-carcinogenic effect s of COPCs over the applicable duration of exposure. Despite quantification, it is believed that the actual pot ential health risks are very low because of the low potential of exposure to chemicals associated with the BalPure ® system. Risks presented below should be c onsidered worst-case estimates.

12.2.5.1 Acceptable risk levels

The risk that is acceptable is very much de pendent on site-specific characteristics that include: the number of people potentially expo sed, the likelihood of exposure, the chemicals driving the risk, and the decisions of risk managers. The increm ental lifetime cancer risk (ILCR) is compared to a range of acceptable probabilities to determine whether the potential risk poses an unacceptable health threat. The US EPA directive, Role of the Baseline Risk Evaluation in Superfund Remedy Selection Decisions (US EPA 1991a), states that action is generally not warranted at a site when the cumulative carcinogenic risk for current and future land use is less than 1 x 10 -4 and the cumulative non-carcinogenic hazard index (HI) is less than 1.0. The level of non-cancer hazard concern increa ses as the HI increa ses above unity, alt hough the two are not linearly related (US EPA 1989). The US EPA uses a potential excess indi vidual lifetime cancer risk of 1 x 10 -6 (1 in 1,000,000) as a point of depa rture for risk management actions.

12.2.5.2 Quantitative estimate of potential risks to STDN BalPure ® t echnicians

Table 35 summarizes the risks quantified for the BalPure ® Technician who may be exposed to concentrated hypochlorous acid (generated concen tration of 0.1%) during maintenance activities or to the 38% sodium bisulfite solution during ch emical resupply. Neither hypochlorous acid nor sodium bisulfite is classifiable with respect to car cinogenicity, therefore cancer risks are not reported below. The non-cancer ha zard of 0.3 is well below the level of concern (1.0); use of personal protective equipment (PPE) and safe chem ical handling procedures will further reduce any potential exposures a nd associated effects.MEPC 60/2/9 ANNEX Page 105

I:\MEPC\60\2-9.doc

Table 35 : Summary of Cancer Risks and Non-cancer Hazards for BalPure ® Technicians

Cancer Risk Non-cancer Hazard

Chemical Ingestio

n Derma l Total Ingestion Dermal Total Hypochlorous Acid NC NC NA 0.0049 0.00032 0.0052 Sodium Bisulfite NC NC NA 0.26 0.017 0.28 TOTAL NA 0.3 NA = not applicable, NC = no criteria

12.2.5.3 Quantitative estimate of potential risks to ship’s crew or dock workers

Table 36 summarizes the risks quantified for ship ’s crew or dock workers who may be exposed to COPCs in ballast water from spray drift. Neither hypochlorous acid nor sodium bisulfite is classifiable with respect to carcinogenicity, ther efore cancer risks are reported for only bromate, THMs, and HAAs. The total estim ated cancer risk of 2 x 10 -9 (2 in 1,000,000,000) is far below the US EPA point of departure of 1 x 10 -6 (1 in 1,000,000). The total non-cancer hazard, 0.00002, is many orders of magnitude below the target level of 1.0.

Table 36 : Summary of Cancer Risks and Non-cancer Hazards for Ship’s Crew/Dock Workers

Cancer Risk Non-cancer Hazard

Chemical Ingestio

n Derma l Total Ingestion Dermal Total Hypochlorous Acid NA NC NA NA 0.0000035 0.0000035

Bromate NA 7.4E11 7.4E-11 NA 0.0000001 8 0.0000001 8

THMs NA 1.6E09 1.6E-09 NA 0.000013 0.000013

HAAs NA 2.0E10 2.0E-10 NA 0.000012 0.000012

Sodium Bisulfite NA NC NA NA 0.0000000 49 0.0000000 49 TOTAL 2E-09 0.00003 NC = no criteria, NA = not applicable

12.2.5.4 Quantitative estimate of potential ri sks to the general public (beachgoers)

Table 37 shows cancer risks to be achgoers who may swim in the vicinity of recently discharged ballast water associated with the BalPure ® system. Neither hypochlorous acid nor sodium bisulfite is classifiable with respect to carcinoge nicity, therefore cancer risks are reported for only bromate, THMs, and HAAs. The total estima ted cancer risk is ve ry low, at 4 x 10 -8 (4 in 100,000,000).MEPC 60/2/9 ANNEX Page 106

I:\MEPC\60\2-9.doc Table 37 : Summary of Cancer Risks for the General Public

Cancer Risk *

Chemical Ingestio

n Derma l Total Hypochlorous Acid NC NC NA

Bromate 4.3E-09 1.0E09 5.3E-09

THMs 1.3E-08 2.2E08 3.5E-08

HAAs 1.8E-09 2.9E09 4.6E-09

Sodium Bisulfite NC NC NA TOTAL 4E-08

* Cancer risk is age-adjusted, assuming 6 years of exposure as a child and 24 years as an adult. NC = No criteria, NA = not applicable. Non-cancer hazards for the general public, which are summarized in Table 38, also are very low; 0.0004 for children and 0.0001 for adults.

Table 38 : Summary of Non-cancer Hazards for General Public

Adult Non-cancer Hazard Child Non-cancer Hazard Chemical Ingestion Dermal Total Ingestion Dermal Total Hypochlorous Acid 0.000039 0.000014 0.000053 0.00018 0.000024 0.00021

Bromate 0.0000021 0.000000 74 0.000002 8 0.000009 6 0.0000013 0.000011

THMs 0.000022 0.000053 0.000075 0.00010 0.000090 0.00019 HAAs 0.000020 0.000048 0.000068 0.000092 0.000082 0.00017

Sodium Bisulfite 0.0000005 4 0.000000 2 0.000000 74 0.000002 5 0.0000003 3 0.0000029

TOTAL 0.0002 0.0006

12.2.5.5 Semi-quantitative evaluation of potential risks from inhalation

The risk estimates reported in Tables 35 through 38 do not include inhalation of COPCs in indoor or outdoor air. While significan t volatilization of CO PCs from ballast water into indoor or outdoor air is not anticipated, some exposure to COPCs in air could occur. The most likely operation that could result in expo sure to COPCs in air is during ballasting, when air in the ballast tanks is vented to the atmo sphere as water is brough t into the tanks. If the ship’s crew is present during this operation, they could be exposed to residual THMs in vent air, since these disinfection by-products are both volatile and hydrophobic. Alt hough concentrations of THMs in vent air have not been measured, worst-case es timates of air concentrations can be made based on maximum ballast water THM concen trations (see sec tion 11.3) and the appropriate Henry’s Law constants (the ratio of the aqueous-phase concentration of a chemical toMEPC 60/2/9 ANNEX Page 107

I:\MEPC\60\2-9.doc its equilibrium partial pressure in the gas phase ). Because vent air will be immediately mixed into the surrounding atmosphere, the air concentra tion estimates are also adjusted for dilution into ambient air by dividing by a factor 10. As a conservativ e evaluation of the inhalation exposure pathway, Table 39 summarizes these data, and compares the estimated air concentrations to U.S. OSHA Permissible Exposure Levels (PELs).

Table 39 : Estimated Concentrations of THMs in Vent Air and Comparisons to PELs

Chemical Maximum Residual Ballast Water Conc. (ug/L) Henry’s Law Constant (unitless) Conc. in Air (mg/m 3 )* Air Exposure

Limit (mg/m 3 )** Air Concentratio

n Exceeds Exposure

Limit? THM Bromoform 1050 0.0218 2.3 5 No Chloroform 53 0.150 0.8 240 No Chlorodibromometh

ane 157 0.0320 0.5 NA NA

Dichlorobromometh

ane 90 0.0654 0.6 NA NA

NA = Not Available * Estimated based on Henry’s Law constant at 25  C, and divided by a factor of 10 for dilution into ambient air. ** U.S. Department of Labor, 2009. Occupa tional Health and Safety Administration, Permissible Exposure Limits (PEL).

Actual air exposures will depend on proximity to vent outlets, atmospheric conditions (wind), as well as the duration of ballasting. Because the TH Ms in vent air would quickly dissipate into the atmosphere, it is anticipated that air exposures will decrease as distance to the ballast water increases; therefore potential air exposures for dock workers and the general public would be even less than those for the ship’s crew. Based on this assessment of THMs in vent air as a result of concentrations in ballast wate r, there is no evidence for the potential of adverse health effects to the ship’s crew, dock work ers, or the general public.

12.2.5.6 Potential risks to consumers of seafood

Chemicals detected in treated ballast water that is discharged may pose a threat to seafood consumers only if they tend to bi oaccumulate (within a trophic leve l, an increase in concentration of a substance in tissues due to uptake from food and sediments) or biomagnify (an increase in the concentration of a substance in the f ood web). As indicated in section 10.1, the bioaccumulation criteria per document MEPC 53/2/1 (or Procedure (G9)) include an octanol-water partition coefficient (Log K ow ) of greater than 3 or a bioconcentration factor greater than 2000. The COPCs identified in this risk as sessment all have Log K ow s less than 3 (Table 16), and therefor e are not expected to persist in the food web.MEPC 60/2/9 ANNEX Page 108

I:\MEPC\60\2-9.doc 12.2.6 Risk assessment conclusions

The health risks to all receptors from the BalPure ® system under normal operating conditions are very small and well within acceptable levels. Even an STDN technician, exposed to concentrated sodium bisulfite during a spill during chemical resupply, and who is wearing no PPE – an unlikely scenario – would have low health risk . Cancer risk and non-cancer hazards for the general public/beachgoers are well below levels of concern; actua l risks to these receptors are expected to be even lower, as they are unlikely to be repeatedly exposed to treated ballast water as assumed in this risk evaluation. Since the risk assessment only evaluated direct exposure via ingestion and dermal contact with chemicals or ba llast water, as these were considered the most relevant pathways, further semi-qualitative eval uation of the inhalation pathway was performed for the volatile THMs. This assessment demons trated that conservativ e concentrations of airborne THMs generated by a release of ballast water were well below occupational exposure limits.

12.3 Risks to the aquatic environment

The data for the substances associated with use of the BalPure ® system indicate that there is a low potential for bioaccumulation, sediment ad sorption, and persistence in the aquatic environment. No effects or risks in the form of secondary (food chain) poisoning or to sediment species are anticipated. As suc h, toxicity presents the only lik ely potential risk for aquatic organisms.

Whole effluent toxicity testing (acute and sub- lethal endpoints) of ne utralized ballast water discharge indicates no apparent toxicity for all species a nd endpoints tested (E(L)C50 and/or NOEC values of ≥ 100% ballast water sample) with th e exception of the marine diatom, Skeletonema (96-hour exposure EC50 of 66.1% sample) (Nautilus, 2009). Less than a two-fold dilution would be required to reach a concentration less than the EC50 for Skeletonema. Certainly, when treated water is discharged to a harbour, the dilu tion would be much greater than two-fold.

To ensure that no risks can be expected from th e oxidant neutralizing agent, further testing was conducted on ballast water neutralized with a large excess of sodium bisulfite (22 mg/L). Testing results were that none of the test acute and/or chronic endpoi nts exhibited a 25% reduction relative to the control. These data suggest that a 3 mg/L treat ment of hypochlorite in seawater with up to 22 mg/L of sodium bisulfite, which is much larger than would normally be applied, would not be expected to exhibit an adverse effect (Nautilus, 2005).

Therefore, the toxicity testing of the treated ballast discharge indi cates that unacceptable risks to the aquatic environment are not e xpected from use of the BalPure ® system.MEPC 60/2/9 ANNEX Page 109

I:\MEPC\60\2-9.doc 13 ASSESSMENT REPORT

13.1 Overview of the data and endpoints

The constituents associated with the BalPure ® system were evaluated for a variety of endpoints, including toxicity, bioaccumulati on, and persistence in the envi ronment. The physical and chemical properties were evaluated to assess proper chemical handling and/or storage in a shipboard setting, and risks such as fire/explosio n. Treated ballast water discharge and the risks of the COPCs to human health were also evaluated.

Because TRO is neutralized prior to discharge, the DBPs potentially present in treated ballast water discharge are the most important to consider for this evaluation. In regards to aquatic toxicity, the existing database for chronic endpoints is limited for many of the THMs and HAAs. As such, the acute database, which is more co mplete, indicates the lowest EC50 was 0.028 mg/L for monochloracetic acid. The evaluation of th e chemical and physical properties of all chemicals relevant to the BalPure ® system suggests that bioaccumulation, sediment adsorption, or persistence is not e xpected to occur. Further, none of the chemicals met all three PBT substance criteria.

Whole effluent discharge toxi city testing has been conduc ted for different water types (fresh water, brackish, and seaw ater) at various Active Substan ce treatment and TOC levels, and at different holding times. Duri ng the most recent studies, and with the exception of one sample for the marine diatom ( Skeletonema ) in brackish water, toxicity was not observed in the Day 5 post-neutralization samples. Although the sensitivity between the taxonomic groups (algae, invertebrate, and fish) varied with the water type (freshwater, brackish, and seawater) tested, fish were the most tolerant species and invertebrates were generally the least tolerant species. Overall, the most sensitive species tested was Daphnia magna (Nautilus, 2009). In all species tested, no toxicity was observed in th e samples tested 5 days after neutralization (Day 5+).

Toxicity tests of treated water discharge fr om studies being done at the Naval Research Laboratory were also conducted. Results for th e chronic toxicity tests are summarized in Table 5, and show that no toxicity was observed with the treated water samples for any species tested. The highest concentra tion with no observed effect ( NOEC) was 100 percent sample (Nautilus, 2007).

Because the BalPure ® process uses an oxidant neutralization step during ballast water discharge, the TRO in the discharge will be non-detectable . Therefore, the discharge and predicted environmental concentration (PEC) for TRO is 0 mg/L.

During pilot studies of the BalPure ® system, the amount of DBPs in treated water discharge were evaluated in three different water types (freshwa ter, brackish water and seawater), at different Active Substance treatments and organic matter le vels. The data is presented in sections 11.1 to 11.3 above. In freshwater, total THMs in treated water ranged from 196.3 μg/L–400 μg/L and total HAAs ranged from 93.7 μg/L–154.7 μg/L. In brackish water, total THMs ranged from 225 μg/L–1099 μg/L and total HAAs ranged from 136.6μg/L–242.2 μg/L. In seawater, total THMs ranged from 33.3 – 121.3 μg/L and total HAAs ranged from 17.8 μg/L–36.8 μg/L. The DBP bromate was also measured in the brackis h and seawater studies. In brackish water, bromate concentrations ranged from <2 μg/L–42 μg/L. In seawater, bromate concentrations ranged from <2 μg/L–23 μg/L.MEPC 60/2/9 ANNEX Page 110

I:\MEPC\60\2-9.doc The DBP concentrations from the studies noted a bove consider analysis of the whole effluent (i.e. discharge concentration) onl y and do not reflect actual envi ronmental concentrations when treated ballast water is released in a harbour where dilution and mixing will occur. Meaningful PEC values will be derived using appropriate emission scenario modelling based on analysis of treated ballast water samples drawn from the fu ll-scale land-based test set-up of the BalPure ® system and will be presented in a future dossier for Final Approval.

Potential human exposures and he alth effects were evaluated qua litatively and quantitatively, when possible. Overall, the risk assessment dete rmined risks to all receptors from the BalPure ® system under normal operating conditions are very small and well within acceptable levels.

A variety of reputable sources we re consulted to gather test data and conduct the assessments required. These sources include, but are not limited to WHO, IPCS, OECD and US EPA. Information was also gathered from published i ndependent research, whic h in most cases is peer-reviewed by others with similar levels of expe rtise. In some cases the test method used was not specified; however if the data set was limited, this data was included to provide at least some information. Due to the primary sources relied upon during the eval uation the level of uncertainty with this assessment is low.

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