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Overview and Status of EPA Collaborations for Detection of Selected Biotoxins in Drinking Water, Soils, and Wipes

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Overview and Status of EPA Collaborations for Detection of Selected Biotoxins in Drinking Water, Soils, and Wipes Overview and Status of EPA collaborations for Detection of Selected Biotoxins in Drinking Water, Soils, and Wipes Matthew L. Magnuson, Ph.D. US Environmental Protection Agency National Homeland Security Research Center Office of Research and Development 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 Office of Research and Development 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 to monitor water quality might be used during other water emergencies and/or clean-up after contamination. Office of Research and Development NHSRC Products • 125 reports and journal articles since 2003 (including classified) • 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.’ Office of Research and Development Overview of Detection of Biotoxins • Laboratory methods – SAM method compendium – Collaborative projects – Future directions Office of Research and Development Laboratory methods • Method development and study – Documentation: Methods and study reports • Methods aim to have 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 (ERLN) • Availability of Methods: Most on-going studies or in clearance – Availability announced on website and we also maintain a list of stakeholders who are specifically informed – Register to get updates at http://www.epa.gov/nhsrc Office of Research and Development Standardized Analytical Methods (SAM) Document • Cornerstone document of Environmental Response Laboratory Network (ERLN) • Identifies chemical, biological and radiological (CBR) agents of concern and preferred analytical methods to be used for ENVIRONMENTAL RESTORATION NOT EMERGENCY RESPONSE • Originally published in 2005, updated annually • SAM 5.0 published in October 2009 • Searchable SAM website: www.epa.gov/sam Office of Research and Development Generic Response Analytical Performance Considerations for Environmental Restoration • Sample Magnitude: 10,000’s of samples for weeks to months. • Multiple Analysis Types: Field and laboratory • Multiple Sample Types: Clinical, Environmental, Forensic • Multiple Sample Matrices: Clinical, food, solids, liquids, etc. Office of Research and Development Analysis needs in Generic Response Timeline Office of Research and Development SAM Revision 5.0 Analytical Methods •Chemical Methods •Radiochemistry Methods •135 analytes •23 analytes •6 sample matrices •5 sample matrices •Sample preparation and •Qualitative determination and determinative methods confirmatory methods •Biotoxin Methods •Pathogen Methods •18 analytes •32 analytes •5 sample matrices •5 sample matrices •Presumptive, confirmatory, •General remediation efficacy added and biological activity methods using spore strips Office of Research and Development SAM Analytical Methods • Sample matrix types – Solid Samples (e.g. soil) – Aqueous liquid samples (e.g. surface water, waste water, etc) – Drinking water samples – Air samples – Wipes (from surfaces) • These are NOT compliance monitoring methods. But the goal is to meet similar DQOs as regulated analytes. Office of Research and Development Method Development Process Goal: All steps between sample collection and data reporting Optimize Instrumentation Determine Best Calibration (chromatography, mass calibrate, (linear/quadratic, internal tune, evaluate instrument stability) standards) Determine Interferences (Are DQOs Determine Preservatives met in various difficult matrices?) (e.g. select antimicrobial & dechlor) YES Determine Does Method Meet Holding Time DQOs? Write Method NO Revise Technical Approach Use in ERLN, WLA Office of Research and Development Adapted from J. Shoemaker and B. Boutin, USEPA/ORD at WQTC 2008 Analytical Target Levels for Methods – Water and Air Risk-based Exposure Guidelines Are Used to Estimate Appropriate Target Levels for Analytical Methods – Risk-Based Criteria to Support Validation of Detection Methods for Drinking Water and Air http://www.epa.gov/nhsrc/pubs/600r08021.pdf Office of Research and Development Detection of Biotoxins in environmental matrices— Biotoxins and SAM • Expert input through SAM Biotoxin Working Group • Proteins – Abrine, Ricin, Bot (A, B, E, F), Shiga 1 & 2, SEA, SEB, SEC • 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 Office of Research and Development Detection of Biotoxins in environmental matrices – Biotoxins and SAM • Analytical techniques – Antibody based detection schemes • ELISA • Lateral flow devices • Various antibody-capture-release detectors (fluorescence, electrochemiluminescence, etc. – Instrumental analysis • Liquid chromatography-mass spectrometry • Gas chromatography-mass spectrometry • others Office of Research and Development 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) Office of Research and Development Previous biotoxin projects Dahlgren Naval Surface Warfare Center • Included both literature review and experimental components. • Develop an improved scientific understanding of the nature of biotoxins potentially used to intentionally contaminate drinking water systems. • Methods of reducing the risks from biotoxin contamination • Target applications: – Decontamination – Threat analysis Office of Research and Development Experimental Approach Designed to reflect possible operational paradigm during water system contamination/decontamination • Different oxidants •chlorine, chloramine, chlorine dioxide, ozone • Fate process •Hydrolysis, Heat inactivation, Coagulation Office of Research and Development Previous biotoxin projects EPA Technology Testing and Evaluation Program (TTEP) Test, evaluate, and report on the performance of homeland security-related technologies • building and outdoor area decontamination • air, water, and wastewater treatment • water security technologies • detectors/monitors for air and water Desired Outcome: Informed decision making by our customers to detect, contain, decontaminate, and manage hazardous chemical, biological, and radiological materials purposefully or accidentally introduced into structures, facilities, drinking water systems, or the environment. Office of Research and Development Reports for various technologies at www.epa.gov/etv • Immunoassay Test Kits for Biotoxins – BADD Anthrax, Botulinum Toxin, and Ricin Immunoassay Test Strips – BioVerify Botulinum Toxin A and Ricin Test Kits and M-Series M1M Analyzer – EzyBot A and EzyBot B Test Kits – QTL Biosensor – RAMP Anthrax, Botulinum Toxin, and Ricin Immunoassay Test Cartridges – BioThreat Alert Anthrax, Botulinum Toxin, and Ricin Immunoassay Test Strips (Tetracore) – Anthrax, Botulinum Toxin, and Ricin Enzyme-Linked Immunosorbent Assay (ELISA, Tetracore) • Rapid Toxicity Testing Systems (included ricin and bot) – AbraTox Kit – Chem-IQ Tox Test Kit – IQ Toxicity Test Rapid Toxicity Testing System – ToxScreen II Rapid Toxicity Testing System (Round 1) – ToxScreen II Test Kit (Round 2) – Toxi-Chromotest – ToxTrak Rapid Toxicity Testing System – BioTox Rapid Toxicity Testing System – POLYTOX Rapid Toxicity Testing System – LuminoTox SAPS Test Kit – Eclox Rapid Toxicity Testing System Office of Research and Development Current biotoxin detection projects Centers for Disease Control and Prevention • The CDC’s Chemical Terrorism Analytical Response Laboratory is currently developing assays for the quantitation of biotoxins in clinical matrices such as urine. These clinical assays are expected to be compatible with drinking water matrices. • LC/MS/MS methods used in chemical laboratory response network (LRN) for clinical samples • Addition of drinking water matrices to experimental design • Stability, extraction, chromatography, MS tuning • High throughput, small samples, automation, IT Office of Research and Development Biotoxins will be extracted from water samples, separated, and analyzed HPLC Solid Phase Extraction Data Analysis Mass Spectrometry Office of Research and Development Features of Adapted Methods • 100-1000 samples/day • Analytes –α , β –Mycotoxins-amanitin –Marine Toxins –TETS – asparagine Biomarkers for ricin and abrin H O 2 N 4-hydroxyproline • O OH Stable Isotopic Internal Standards O N cysteine NH HN O HO O 4,5-dihydroxyisoleucine O • S HN Availability: Fall 2010 HN OH glycine NH HN O O HO H N 4-hydroxytryptophan isoleucine O O Exact Mass: 918.35 NH glycine Office of Research and Development Native Labeled ISTD Current Projects Centers for Disease Control and Prevention
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