Water and Wastewater Analysis Following Catastrophic Events

Matthew L. Magnuson & Hiba S. Ernst US Environmental Protection Agency/Office of Research and Development National Homeland Security Research Center/Water Infrastructure Protection Division

National Environmental Monitoring Conference San Antonio, TX, August 5-8, 2013 Presentation Overview

• Lab analysis during catastrophic events - Is there a difference? • What is being done? – Lab Networks – Pre-selected and verified methods – Biotoxin research projects – Chemistry research projects

2 NHSRC Mission

To conduct research and develop scientific products that improve the capability of the Agency to carry out its homeland security responsibilities

ADVANCING OUR NATION’S SECURITY THROUGH SCIENCE EPA Homeland Security Roles

• Protecting water and wastewater infrastructure • Indoor and outdoor clean-up following attack or natural disaster . can use millions of gallons of water . can result in even more contaminated wastewater • Development of a nationwide laboratory network • Reducing vulnerability of chemical & hazardous materials • Cyber security NHSRC Research Projects

Homeland Security

Multi Use Cross Agency

“Normal” Environmental Operations

Many homeland security practices may also benefit day to day operation. For example, emerging analytical techniques useful for water emergencies and/or clean-up after contamination might also be useful for monitoring water quality. Water Security & Sustainability

6 Water Security Threats

Water Distribution Networks

• Multiple points of access over thousands of miles of pipe • Vulnerable to insider, physical, and cyber threats • Vulnerable to intentional or accidental contamination (e.g., injection at hydrant or cross-connection) • Contamination might not be detected until people are sick or in the hospital • Intelligence information on adversary capabilities 7 Why Waste Water?

• Wide area contamination release could affect water sources. • Water can become contaminated not only from the initial release, but also as a result of decontamination activities. • Millions of gallons of water may be used for decon. • Contaminated water may enter the storm or wastewater collection system, impacting the wastewater treatment plant and receiving waters.

8 CBR Decontamination and Consequence Management Paradigm

Analysis is key

Contamination Characteristics • Persistence • Chemical • Physical

Waste Spread of Management Contamination • Pre-decon • Dispersion • Post-decon • Resuspension • Treatment/ • Tracking Handling Decon Method • Efficacy • Engineering Possible Numbers of Samples from an Incident

10 Meeting Lab Throughput

Potential approaches • High throughput sampling techniques and laboratory analysis techniques, including automated or rapid method capabilities

• Methods which can be applied to multiple labs by Environmental Response Laboratory Network (ERLN) and Water Laboratory Alliance

11

Selected Analytical Methods (SAM)

SAM 2012 Published: July 2012

Chemical Methods 142 analytes 5 matrices Pathogen Methods 31 analytes 4 matrices Radiochemical Methods 25 analytes 6 matrices Biotoxin Methods 18 analytes 5 matrices

12 www.epa.gov/sam

Laboratory Method Development

• Methods aim to have data quality objectives (DQOs) fit for their intended use by – EPA/Water Security Division through the Water Laboratory Alliance – EPA/Office of Emergency Management through the Environmental Response Laboratory Network

• Availability of Method and Study Reports – Availability announced on website, and we also maintain a list of stakeholders who are specifically informed – Register at http://www.epa.gov/nhsrc NHSRC Research Products

• Results presented many other ways—stakeholder meetings, symposia, workshops, etc. • Products and research plans receive rigorous quality reviews

Most scientists regarded the new streamlined peer-review process as ‘quite an improvement.’ Method Development in Water Matrices (Biotoxins and Chem) Method Development for Biotoxins

• Protein – Abrin, Ricin, Botulinum neurotoxin (A, B, E, F), Shiga 1 & 2, Staphylococcal Enterotoxin (A, B, C) • Small molecule – Aflatoxin B1, α-Amanitin, anatoxin-a, brevetoxin B, α-Conotoxin, Cylindrospermopsin, Diacetoxyscirpenol, microcystins (LA, LR, LW, RR, YR), picrotoxin, saxitoxins, T2 mycotoxin, tetrodotoxin

Detection of Biotoxins

• Analytical techniques – Antibody-based detection schemes • Enzyme Linked Immunosorbant Assay (ELISA) • Lateral flow devices • Various antibody-capture-release detectors (fluorescence, electrochemiluminescence, etc.) – Instrumental analysis • Liquid chromatography-mass spectrometry • Gas chromatography-mass spectrometry • others

Meeting Throughput Requirements

Multi-tier analysis approach • Environmental restoration hopefully is effective, so while there are many samples, fewer will be positive after initial decontamination activities • Samples initially subjected to higher throughput screening methods • Followed by analysis of selected samples with lower throughput, but more definitive, techniques • Application approach of techniques listed will appear in future version of SAM (e.g. see techniques of ricin in table below)

Biotoxin Detection Projects

Centers for Disease Control and Prevention • Quantitation of biotoxins via methods adapted from clinical matrices such as urine • LC/MS/MS methods, as used in chemical laboratory response network (LRN) for clinical samples – Adaptation to drinking water matrices – Stability, extraction, chromatography, MS tuning – High throughput, small samples, automation, IT • Enzyme Linked Immunosorbant Assay (ELISA) for Botulinum toxin in clinical samples and food adapted to water • Endopep-MS (enzymatic activity) method adapted to water samples • Ability of the ultrafiltration (UF) technique to concentrate botulinum toxin in water samples

Features of Adapted Methods

-amanitin e lin pro xy dro -hy 4 e cin OH leu so ine yi ag rox ar yd asp O ih O 5-d HO 4, H2N N O OH

O NH HN O cysteine O S HN HN HN an ph pto try • 100-1000 samples/day xy ro yd O 4-h

e NH cin HO O gly H O N NH

e O cin • Analytes gly e cin leu iso – Alpha amanitin Exact Mass: 918.35 – TETS – Biomarkers for ricin and abrin • Stable Isotopic Internal Standards Native

Labeled ISTD

Method Development for Chemicals Verification of Chemicals in Existing Method

• Use methods as written, for lab familiarity • Understand performance, demonstrate all steps including QC • Examples . Semi-volatiles (e.g. pesticides) . CWA transformation products . Metals

22 Immunomagnetic Scavenging and LC/MS Detection of VX in Water

1) Conjugation of antibody to beads 4) LC/MS/MS Analysis

2) Binding of antibody to BuChE

3) Expose 3) Protein to water digestion

Magnetic Bead Antibody BuChE Peptides IMSc LC-MS/MS Method for Detection of VX in Water

• Method sensitivity down to the ppt level – Calculated method detection limit = 5.6 ng/L – Minimum reportable level = 25 ng/L – Small sample size (100 mL) • Can be used to analyze up to 500 samples per day • Low concentrations of VX can be detected in preserved tap water 91 days after spiking – Suggests applicability of this method for determining water contamination with VX and verifying environmental remediation

24 Wastewater Treatment Process

25 Analysis for System Studies

Degradation of contaminants by activated sludge Example contaminants: . EMPA (ethyl methyl phosphonic acid) : Environmental and decon product of VX, determined by LC/MS/MS . Malathion: Simulant for VX, determined by GC/MS . Evaluated total suspended solid concentration and contact time Conclusions: • If nitrifiers are active, they might degrade the analyte of concern (less than 25 percent) • EMPA, malathion, and similarly sorbed and biodegraded compounds may pass through an activated sludge wastewater treatment plant largely unchanged 26

Future Directions

• Extension to “All-hazard” catastrophes –natural disasters, industrial accidents, etc • Analysis approaches for other matrices • Additional biotoxins and chemicals • Refinement of existing methods • Application-focused studies (decontamination, system operations, etc.) • Enhanced collaborations – Federal (e.g. recent EPA, DHS, and DOD agreement) – Other: maybe YOU?!

Acknowledgements

• Centers for Disease Control and Prevention • Naval Surface Warfare Center – Rudy Johnson, Ph.D. – Elaine Strauss, Ph.D. – Jennifer Links, Ph.D. – Wynn Vo – Stephen Morse, Ph.D. – CT method – Andrew Slaterbeck, Ph.D. development group – Bradford Gutting, Ph.D.

• Food Safety Inspection Service • Environmental Protection Agency – Mark Campbell, Ph.D. – Michelle Burgess, Ph.D., EPA – Marcus Head, Ph.D. – Jim Jones, Ph.D. – Sanjiv Shah, Ph.D., NHSRC – Anne Hurley, DVM, MPH – Gene Rice, Ph.D., NHSRC – Hiba Ernst, Ph.D., NHSRC • US Air Force Institute of Technology – Alan Lindquist, Ph.D., NHSRC – Maj. LeeAnn Racz, Ph.D. – Frank Schaefer, Ph.D., NHSRC – Lt. Allen Janeckso, M.S. – Maj. Edward Walters, M.S. – Vince Gallardo – Tonya, Nichols, Ph.D., NHSRC

Acknowledgements

SAM 2012 Matthew Magnuson, Ph.D. Published: July 2012 Acting Associate Director/Chemist Water Infrastructure Protection Division [email protected]; 513-569-7321

Co-authors/Collaborators: Romy Campisano Erin Silvestri Stuart Willison

U.S. EPA, National Homeland Security Research Center

DISCLAIMER: The U.S. EPA through its Office of Research and Development partially funded the research described in this presentation. It has been reviewed by the Agency but does not Website: www.epa.gov/sam necessarily reflect the Agency’s views. No official endorsement should be inferred. EPA does not endorse the purchase or sale 29 of any commercial products or services.