&EPA
United States
Environmental Protection
Agency
Standardized Analytical Methods
for Environmental Restoration
Following Homeland Security
Events
REVISION 3.1
Office of Research and Development
National Homeland Security Research Center
i
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EPA/600/R-07/136
November 15, 2007
Standardized Analytical Methods for
Environmental Restoration following
Homeland Security Events
Revision 3.1
November 15, 2007
Prepared by
Computer Sciences Corporation
Alexandria, VA 22304-3540
Prepared under
EPA Contract No. EP-W-06-046
Prepared for
National Homeland Security Research Center
United States Environmental Protection Agency
Office of Research and Development
Cincinnati, OH 45268
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Disclaimer
Disclaimer
The U.S. Environmental Protection Agency through its Office of Research and Development funded and
managed the research described here under Contract EP-W-06-046 with Computer Sciences Corporation
(CSC). This document has been subjected to EPA's peer and administrative review and has been
approved for publication as an EPA document.
Mention of trade names or commercial products in this document or in the methods referenced in this
document does not constitute endorsement or recommendation for use.
Questions concerning this document or its application should be addressed to:
Oba Vincent
National Homeland Security Research Center
Office of Research and Development (163)
U.S. Environmental Protection Agency
26 West Martin Luther King Drive
Cincinnati, OH 45268
(513)569-7456
yjnrent.oba@epa.gov
SAM Revision 3.1 iii November 15, 2007
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Use of This Document
Use of This Document
The information contained in this document represents the latest step in an ongoing National
Homeland Security Research Center effort to provide standardized analytical methods for use by
laboratories (e.g., EPA contract laboratories) tasked with performing confirmatory analyses of
environmental samples following a homeland security incident. Although at this time, some of the
methods listed have not been fully validated for a particular analyte (e.g., analytes not explicitly
identified in the method) or sample type, the methods are considered to contain the most
appropriate currently available techniques. Unless a published method that is listed in this
document states specific applicability to the analyte/sample type for which it has been selected, it
should be assumed that method testing is needed, and adjustments may be required to accurately
account for variations in analyte/sample type characteristics, environmental samples, and target
risk levels. Many of the target analytes listed in this document have only recently become an
environmental concern. EPA is actively pursuing development and validation of Standard
Analytical Protocols (SAPs) based on the methods listed, including optimization of procedures for
measuring target compounds. In those cases where method procedures are determined to be
insufficient for a particular situation, EPA will provide guidance regarding appropriate actions. This
will be an ongoing process as EPA will strive to establish a consistent level of validation for all
listed analytes.
SAM Revision 3.1 iv November 15, 2007
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Foreword
Foreword
The U.S. Environmental Protection Agency (EPA) is charged by Congress with protecting the Nation's
land, air, and water resources. Under a mandate of national environmental laws, EPA strives to formulate
and implement actions leading to a compatible balance between human activities and the ability of natural
systems to support and nurture life. To meet this mandate, EPA's research program is providing data and
technical support for solving environmental problems today and building a scientific base necessary to
manage our ecological resources wisely, understand how pollutants affect our health, and prevent or
reduce environmental risks in the future.
The National Homeland Security Research Center (NHSRC) is EPA's center for conducting research to
facilitate protection and decontamination of structures and water infrastructure subject to chemical,
biological, or radiological (CBR) terror attacks. NHSRC's research is designed to provide appropriate,
effective, and validated technologies, methods, and guidance to understand the risks posed by CBR agents
and to enhance our ability to detect, contain, and clean up in the event of an incident involving such
agents. This document is intended to provide guidance for selecting methods that have a high likelihood
of ensuring analytical consistency when laboratories are faced with a large scale environmental
restoration crisis. At the same time, the document can be used as a tool to identify analytes that require
further methods development and verification to ensure desired performance. NHSRC will also provide
direct technical assistance to response personnel in the event of a CBR attack, as well as provide related
interagency liaisons.
This publication has been produced as part of the NHSRC's long-term research plan. It is published and
made available by EPA's Office of Research and Development to assist the user community and to link
researchers with their clients.
Jonathan G. Herrmann, Director
National Homeland Security Research Center
SAM Revision 3.1 v November 15, 2007
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Foreword
SAM Revision 3.1 vi November 15, 2007
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Abbreviations and Acronyms
Abbreviations and Acronyms
AEM Applied and Environmental Microbiology
AGI sampler All Glass Impinger Sampler
2-Am-DNT 2-Amino-4,6-dinitrotoluene
4-Am-DNT 4-Amino-2,6-dinitrotoluene
Amp-ELISA Amplified-Enzyme-Linked Immunosorbent Assay
APHA American Public Health Association
AOAC AOAC International (formerly the Association of Official Analytical Chemists)
ASTM ASTM International (formerly the American Society for Testing and Materials)
AWWA American Water Works Association
BCM Biosynth Chromogen Medium
BGMK Buffalo Green Monkey Kidney
BS Bismuth Sulfite
BLEB Buffered Listeria Enrichment Broth
BSA Bovine Serum Albumin
BSL Biosafety Level
°C Degrees Celsius
Campy-BAC Campylobacter-Brucella agar base with sheep blood and antibiotics
CAS RN Chemical Abstracts Service Registry Number
CBR Chemical, Biological, or Radiological
CCID Coordinating Center for Infectious Diseases
CDC Centers for Disease Control and Prevention
CFR Code of Federal Regulations
CFSAN Center for Food Safety and Applied Nutrition
CPU Colony Forming Unit
CLP Contract Laboratory Program
CoV Coronavirus
CPE Cytopathic Effect
cps counts per second
CS Tear gas; Chlorobenzylidene malonitrile
CVAA Cold Vapor Atomic Absorption or 2-Chlorovinylarsonous acid
CVAFS Cold Vapor Atomic Fluorescence Spectrometry
DAPI 4',6-diamidino-2-phenylindole
DAS Diacetoxyscipenol
DAS-HG-HAS Diacetoxyscipenol Hemiglutarate Human Serum Albumin
DAS-HS-HRP Diacetoxyscipenol Hemisuccinate Horseradish Peroxidase Conjugate
DBPR Division of Bioterrorism Preparedness and Response
DHS U.S. Department of Homeland Security
DIC Differential Interference Contrast
DIG-ELISA Digoxigenin Labeled Enzyme-Linked Immunosorbent Assay
DIMP Diisopropyl methylphosphonate
DNA Deoxyribonucleic Acid
3,5-DNA 3,5-Dinitroaniline
1,3-DNB 1,3-Dinitrobenzene
2,4-DNPH 2,4-Dinitrophenylhydrazine
2,4-DNT 2,4-Dinitrotoluene
2,6-DNT 2,6-Dinitrotoluene
DoD U.S. Department of Defense
DOE U.S. Department of Energy
DOT U.S. Department of Transportation
DPD N, N-diethyl-p-phenylenediamine
EA2192 Diisopropylaminoethyl methylthiophosphonate
SAM Revision 3.1
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Abbreviations and Acronyms
BCD Electron Capture Detector
ECL Electrochemiluminescence
ED Electron Diffraction or Ethyldichloroarsine
EDEA N-Ethyldiethanolamine
EDL Estimated Detection Limit
EDTA Ethylenediaminetetraacetic acid
EDXA Energy Dispersive X-ray Analysis
EEB EHEC Enrichment Broth
EHEC Enterohemorrhagic Escherichia coll
EIA Enzyme Immunoassay
ELCD Electrolytic Conductivity Detector
ELISA Enzyme-Linked Immunosorbent Assay
EMC Emission Measurement Center
EMJH Ellinghausen-McCullough Johnson Harris Formulation
EML Environmental Measurements Laboratory
EMMI Environmental Monitoring Methods Index
EMPA Ethyl methylphosphonic acid
EMSL Environmental Monitoring and Support Laboratory
EPA U.S. Environmental Protection Agency
EQL Estimated Quantitation Limit
ESI Electrospray lonization
ETV Environmental Technology Verification
FA Fluorescence Assay
FBI Federal Bureau of Investigation
FDA Food and Drug Administration
FEMS Federation of European Microbiological Societies
FGI Fluorescein derivative of Conus geographus a-conotoxin
FID Flame lonization Detector
FITC Fluorescein Isothiocyanate
FRET Fluorescence Resonance Energy Transfer
FSIS Food Safety and Inspection Service
Furadan Carbofuran
GA Tabun
GC Gas Chromatograph or Gas Chromatography
GC-MS Gas Chromatograph/Mass Spectrometer or Gas Chromatography/Mass Spectrometry
GD Soman
GE 1-Methylethyl ester ethylphosphonofluoridic acid or Genome Equivalent
Ge(Li) Germanium (Lithium)
GF Cyclohexyl sarin
GFAA Graphite Furnace Atomic Absorption Spectrophotometer or Graphite Furnace Atomic
Absorption Spectrophotometry
GITC Guanidinium Isothiocyanate
GTC Guanidinium Thiocyanate
HA Hemagglutinin
HAdV Human Adenoviruses
HASL Health and Safety Laboratory, currently known as Environmental Measurements
Laboratory (EML)
HAV Hepatitis A Virus
HCoV Human Coronavirus
HCT Human Ileocecal Adenocarcinoma
HEV Hepatitis E Virus
HECD Hall Electrolytic Conductivity Detector
HEPA High Efficiency Particulate Air (Filter)
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Abbreviations and Acronyms
HD Sulfur mustard / mustard gas; bis(2-chloroethyl) sulfide
HHA Hand-held Assay (e.g., immunochromatographic test device)
HHS Health and Human Services
HMTD Hexamethylenetriperoxidediamine
HMX Octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine
HN-1 Nitrogen mustard 1; bis(2-chloroethyl)ethylamine
HN-2 Nitrogen mustard 2; 2,2'-dichloro-N-methyldiethylamine N,N-bis(2-
chloroethyl)methylamine
HN-3 Nitrogen mustard 3; tris(2-chloroethyl)amine
HPGe High Purity Germanium
HPLC High Performance Liquid Chromatograph or High Performance Liquid Chromatography
HPLC-FL High Performance Liquid Chromatograph - Fluorescence
HPLC-MS High Performance Liquid Chromatograph - Mass Spectrometer
HPLC-PDA High Performance Liquid Chromatography - Photodiode Array
HPLC-vis High Performance Liquid Chromatography - visible
1C Ion Chromatograph or Ion Chromatography
ICC Integrated Cell Culture
ICC/RT-PCR Integrated Cell Culture/Reverse Transcriptase Polymerase Chain Reaction
ICP Inductively Coupled Plasma
ICP-AES Inductively Coupled Plasma - Atomic Emission Spectrometry
ICP-MS Inductively Coupled Plasma - Mass Spectrometry
ICR Information Collection Rule or Information Collection Request (depending on context)
ID50 A dose which would be infectious to 50% of the population
IMPA Isopropyl methylphosphonic acid
IMS Immunomagnetic Separation
INCHEM INCHEM is a means of rapid access to internationally peer reviewed information on
chemicals commonly used throughout the world, which may also occur as contaminants
in the environment and food. It consolidates information from a number of
intergovernmental organizations whose goal it is to assist in the sound management of
chemicals, http://www.inchem.org/
IO Inorganic
IRIS Integrated Risk Information System (U.S. EPA)
ISO International Organization for Standardization
ISE Ion Specific Electrode
K-D Kuderna-Danish
L-l Lewisite 1; 2-Chlorovinyldichloroarsine
L-2 Lewisite 2; bis(2-Chlorovinyl)chloroarsine
L-3 Lewisite 3; tris(2-Chlorovinyl)arsine
LB-M Lim Benyesh-Melnick
LC Liquid Chromatograph
LC/APCI-MS Liquid Chromatography Atmospheric Pressure Chemical lonization Mass Spectrometry
LC/ESI-MS Liquid Chromatography Electrospray lonization Mass Spectrometry
LC-MS-MS Liquid Chromatography Tandem Mass Spectrometry
LIA Lysine Iron Agar
LLD Lower Limit of Detection
LOD Limit of Detection
LRN Laboratory Response Network
LSE Liquid/Solid Extraction
M Molar
Mab Monoclonal Antibody
MARLAP Multi-Agency Radiological Laboratory Analytical Protocols
MDL Method Detection Limit
MLD Minimum Lethal Dose
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Abbreviations and Acronyms
MPA Methylphosphonic acid
MPN Most Probable Number
MS Mass Spectrometer or Mass Spectrometry or Matrix Spike
MSB Matrix Spike Duplicate
MSRV Modified Semisolid Rappaport-Vasiliadis
MTBE Methyl fert-butyl ether
MW Molecular Weight
NA Not Applicable
Nal(Tl) Thallium-Activated Sodium Iodide
NB Nitrobenzene
NBD chloride 7-Chloro-4-nitrobenzo-2-oxa-l,3-diazol
NCPDCID National Center for the Prevention, Detection, and Control of Infectious Diseases
NEMI National Environmental Methods Index
NERL-CI National Exposure Research Laboratory-Cincinnati
NG Nitroglycerine
NH3-N Ammonia Nitrogen
NHSRC National Homeland Security Research Center
NIOSH National Institute for Occupational Safety and Health
NIST National Institute of Standards and Technology
nM Nanomolar
NNSA National Nuclear Security Administration
NoV Norovirus
NPD Nitrogen-phosphorus Detector
NOS Not Otherwise Specified
NRC National Research Council
nS nano Siemens
2-NT 2-Nitrotoluene
3-NT 3-Nitrotoluene
4-NT 4-Nitrotoluene
NTIS National Technical Information Service
NTU Nephelometric Turbidity Units
OAQPS Office of Air Quality Planning and Standards
OEM U.S. EPA Office of Emergency Management
ONPG ort/20-Nitrophenyl-p-D-galactopyranoside
ORAU Oak Ridge Associated Universities
ORIA U.S. EPA Office of Radiation and Indoor Air
ORISE Oak Ridge Institute of Science and Education
ORD U.S. EPA Office of Research and Development
ORF Open Reading Frame
OSWER U.S. EPA Office of Solid Waste and Emergency Response
OSHA Occupational Safety and Health Administration
OW U.S. EPA Office of Water
OXA Oxford Medium
PAHs Poly cyclic Aromatic Hydrocarbons
PBS Phosphate Buffered Saline
PCDDs Polychlorinated Dibenzo-p-dioxins
PCDFs Polychlorinated Dibenzofurans
pCi Picocuries
PCR Polymerase Chain Reaction
PEL Permissible Exposure Limit
PETN Pentaerythritol tetranitrate
PFE Pressurized Fluid Extraction
PFIB Perfluoroisobutylene
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Abbreviations and Acronyms
PID50 50% Pig Infectious Dose
PMPA Pinacolyl methyl phosphonic acid
PubMED PubMED is a service of the U.S. National Library of Medicine (www.pubmcd.gov).
containing citations from scientific journals
PV Partially Validated
QA Quality Assurance
QC Quality Control
qPCR Qualitative Polymerase Chain Reaction
RDX Hexahydro-1,3,5 -trinitro-1,3,5 -triazine
RNA Ribonucleic Acid
RP-HPLC Reversed-Phase High Performance Liquid Chromatography
rRNA Ribosomal Ribonucleic Acid
RTECS Registry of Toxic Effects of Chemical Substances
RT-PCR Reverse Transcription-Polymerase Chain Reaction
SAED Select Area Electron Diffraction
SAM Standardized Analytical Methods for Environmental Restoration following Homeland
Security Events
SAP Standardized Analytical Protocol
SARS Severe Acute Respiratory Syndrome
SaV Sapovirus
SM Standard Methods for the Examination of Water and Wastewater
SPE Solid-Phase Extraction
spp. Species (plural)
STEC Shiga-toxigenic E. coll
STEL Short Term Exposure Limit
STX Saxitoxin
Stx Shiga Toxin
Stx-1 Shiga Toxin Type 1
Stx-2 Shiga Toxin Type 2
SW Solid Waste
TATP Triacetone triperoxide
TBD To Be Determined
TCBS Thiosulfate Citrate Bile Salts Sucrose
TC SMAC Tellurite Cefixime Sorbitol MaConkey Agar
TCLP Toxicity Characteristic Leaching Procedure
TDG Thiodiglycol
TEA Triethanolamine
TEM Transmission Electron Microscope or Microscopy
Temik Aldicarb
TETR-PCR Touchdown Enzyme Time Release-Polymerase Chain Reaction
THF Tetrahydrofuran
1,3,5-TNB 1,3,5-Trinitrobenzene
2,4,6-TNT 2,4,6-Trinitrotoluene
TOFMS Time of Flight Mass Spectrometry
TOXNET Toxicology Data Network
TRF Time Resolved Fluorescence
TRU Transuranic
TSB Tryptic Soy Broth
TTX Tetrodotoxin
TSAye Trypticase Soy Agar with yeast extract
TSC Tryptose-Sulfite Cycloserine
TSI Triple Sugar Iron
TSP Thermospray
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Abbreviations and Acronyms
TTN Technical Transfer Network
TWA Total Weighted Average
USDA U.S. Department of Agriculture
USGS U.S. Geological Survey
UV Ultraviolet
VCSB Voluntary Consensus Standard Body
VE Phosphonothioic acid, ethyl-, S-(2-(diethylamino)ethyl) O-ethyl ester
VEE Venezeulan Equine Encephalitis
VM Phosphonothioic acid, S-(2-(diethylamino)ethyl) O,O-diethyl ester
VOCs Volatile Organic Compounds
VOA Volatile Organic Analysis
VX O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate
WEF Water Environment Federation
WHO World Health Organization
XLD Xylose Lysine Desoxycholate
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Acknowledgments
Acknowledgments
The contributions of the following persons and organizations to the development of this document are
gratefully acknowledged:
United States Environmental Protection Agency
• Office of Research and Development, National Homeland Security Research Center
Joan Bursey
Kathy Hall
Romy Lee
Alan Lindquist
Matthew Magnuson
Tonya Nichols
Ben Packard
Eugene Rice
Rob Rothman
Frank Schaefer
Sanjiv Shah
Ramona Sherman
Erin Silvestri
Oba Vincent
Charlena Yoder
• Office of Research and Development, National Exposure Research Laboratory (NERL), Las
Vegas
Don Betowski (Environmental Sciences Division)
Christian Daughton (Environmental Sciences Division)
Jane Denne (Environmental Sciences Division)
Michael Hiatt (Environmental Sciences Division)
Tammy Jones-Lepp (Environmental Sciences Division)
Stephen Pia (Environmental Sciences Division)
Lee Riddick (Environmental Sciences Division)
Charlita Rosal (Environmental Sciences Division)
Brian Schumacher (Environmental Sciences Division)
Wayne Sovocool (Environmental Sciences Division)
Katrina Varner (Environmental Sciences Division)
John Zimmerman (Environmental Sciences Division)
Kim Rogers (Human Exposure and Atmospheric Sciences Division)
Jeannette Van Emon (Human Exposure and Atmospheric Sciences Division)
• Office of Research and Development, National Exposure Research Laboratory (NERL),
Cincinnati
James Owens (Microbiological and Chemical Exposure Assessment Research Division)
Gerard Stelma (Microbiological and Chemical Exposure Assessment Research Division)
Ann Grimm (Microbiological and Chemical Exposure Assessment Research Division)
• Office of Solid Waste and Emergency Response (OSWER)
Barry Lesnik (Office of Solid Waste)
Michael Johnson (Office of Superfund Remediation and Technology Innovation)
Lawrence Kaelin (Office of Emergency Management, National Decon. Team)
Michael Ottlinger (Office of Emergency Management, National Decon. Team)
Terry Smith (Office of Emergency Management)
Shen Yi-yang (Office of Emergency Management)
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Acknowledgments
• Office of Air and Radiation, Office of Radiation and Indoor Air
Lindsey Bender (Radiation Protection Division)
George Dilbeck (National Air and Radiation Environmental Laboratory)
Dennis Farmer (Radiation and Indoor Environments National Laboratory)
John Griggs (National Air and Radiation Environmental Laboratory)
Daniel Mackney (National Air and Radiation Environmental Laboratory)
David Musick (Radiation and Indoor Environments National Laboratory)
• Office of Water, Office of Ground Water and Drinking Water
Jessica Pulz (Water Security Division)
• Office of Prevention, Pesticides, and Toxic Substances, Office of Pesticide Programs
Yaorong Qian (Biological and Economic Analysis Division)
• Office of Enforcement and Compliance Assurance, Office of Criminal Enforcement,
Forensics and Training
Larry Strattan (National Enforcement Investigations Center)
• EPA Regions
John Curry (Region 3)
Joe Dorsey (Region 3)
Diane Gregg (Region 6)
Adrienne Greenlee (Region 7)
Stephanie Harris (Region 10)
Peggy Knight (Region 10)
Ed O'Neill (Region 6)
Peter Philbrook (Region 1)
Steve Reimer (Region 10)
Dennis Revell (Region 4)
Norman Rodriguez (Region 7)
Sue Warner (Region 3)
Laura Webb (Region 7)
Wayne Whipple (Region 5)
Larry Zintek (Region 5)
United States Centers for Disease Control and Prevention (CDC)
Kevin Ashley (National Institute for Occupational Safety and Health)
Raymond Biagini (National Institute for Occupational Safety and Health)
Erin Black (National Center for Infectious Diseases)
Cheryl Bopp (National Center for Infectious Diseases)
Jay Gee (National Center for Infectious Diseases)
Vince Hill (National Center for Infectious Diseases)
Rudolph Johnson (National Center for Environmetnal Health)
Richard Kellogg (National Center for Infectious Diseases)
Gene Kennedy (National Institute for Occupational Safety and Health)
Stephen A. Morse (National Center for Infectious Diseases)
John Snawder (National Institute for Occupational Safety and Health)
Richard Wang (National Center for Environmental Health)
United States Department of Agriculture
Jim Trapp
Jim Trout
Paul Zimba
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Acknowledgments
United States Department of Commerce
Peter Moeller (National Oceanic & Atmospheric Administration)
United States Food and Drug Administration
Jennifer Brzezinski
David Craft
Eric Garber
United States Department of Homeland Security
Anna Berne (Environmental Measurements Laboratory)
United States Department of Defense
Johnathan Kiel (U.S. Air Force)
United States Department of the Army
Ronald Swatski (Center for Health Promotion and Preventive Medicine)
Sina Bavari (Medical Research Institute of Infectious Diseases, USAMRIID)
Alan Hewitt (U.S. Army Corps of Engineers)
Mark Poli (Medical Research Institute of Infectious Diseases, USAMRIID)
State Agencies
Rick Bokanyi (Ohio Department of Health)
David Degenhardt (Wisconsin State Lab of Hygiene)
Robert Glowacky (Virginia State Department of General Services)
Ted Haigh (Oregon Department of Environmental Quality)
Rebecca Hoffman (Wisconsin State Lab of Hygiene)
Christopher Retarides (Virginia State Department of General Services)
Tom York (Virginia State Department of General Services)
Sandia National Laboratory
Gary Brown
Hamilton Sundstrand
Richard Trubey
Computer Sciences Corporation
Eric Boring
Danielle Carter
Joan Cuddeback
Melody Jensen
Larry Umbaugh
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Acknowledgments
SAM Revision 3.1 xvi November 15, 2007
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Table of Contents
Standardized Analytical Methods for Environmental Restoration
following Homeland Security Events
Revision 3.1
November 15, 2007
Contents
Disclaimer iii
Use of This Document iv
Foreword v
Abbreviations and Acronyms vii
Acknowledgments xiii
Section 1.0: Introduction 1
Section 2.0: Background 3
Section 3.0: Scope and Application 7
Section 4.0: Points of Contact 11
Section 5.0: Selected Chemical Methods 13
5.1 General Guidance 14
5.1.1 Standard Operating Procedures for Identifying Chemical Methods 14
5.1.2 General Quality Control (QC) Guidance for Chemical Methods 27
5.1.3 Safety and Waste Management 29
5.2 Method Summaries 29
5.2.1 EPA Method 8: Determination of Sulfuric Acid and Sulfur Dioxide Emissions from
Stationary Sources 29
5.2.2 EPA Method 200.8: Determination of Trace Elements in Waters and Wastes by Inductively
Coupled Plasma-Mass Spectrometry 30
5.2.3 EPA Method 245.2: Mercury (Automated Cold Vapor Technique) 31
5.2.4 EPA Method 252.2: Osmium (Atomic Absorption, Furnace Technique) 31
5.2.5 EPA Method 300.1, Revision 1.0: Determination of Inorganic Anions in Drinking Water by
Ion Chromatography 32
5.2.6 EPA Method 335.4: Determination of Total Cyanide by Semi-Automated Colorimetry 32
5.2.7 EPA Method 350.1: Nitrogen, Ammonia (Colorimetric, Automated Phenate) 33
5.2.8 EPA Method 507: Determination of Nitrogen- and Phosphorus-Containing Pesticides in
Water by Gas Chromatography with a Nitrogen-Phosphorus Detector 33
5.2.9 EPA Method 524.2: Measurement of Purgeable Organic Compounds in Water by Capillary
Column Gas Chromatography / Mass Spectrometry 34
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Table of Contents
5.2.10 EPA Method 525.2: Determination of Organic Compounds in Drinking Water by Liquid-
Solid Extraction and Capillary Column Gas Chromatography / Mass Spectrometry 34
5.2.11 EPA Method 531.2: Measurement of N-Methylcarbamoyloximes and N-Methylcarbamates
in Water by Direct Aqueous Injection HPLC with Postcolumn Derivatization 35
5.2.12 EPA Method 549.2: Determination of Diquat and Paraquat in Drinking Water by Liquid-
Solid Extraction and High-Performance Liquid Chromatography with Ultraviolet Detection
35
5.2.13 EPA Method 551.1: Determination of Chlorination Disinfection Byproducts, Chlorinated
Solvents, and Halogenated Pesticides/Herbicides in Drinking Water by Liquid-Liquid
Extraction and Gas Chromatography with Electron-Capture Detection 36
5.2.14 EPA Method 614: The Determination of Organophosphorus Pesticides in Municipal and
Industrial Wastewater 36
5.2.15 EPA Method 3031 (SW-846): Acid Digestion of Oils for Metals Analysis by Atomic
Absorption or ICP Spectrometry 37
5.2.16 EPA Method 3050B (SW-846): Acid Digestion of Sediments, Sludges, and Soils 37
5.2.17 EPA Method 3520C (SW-846): Continuous Liquid-Liquid Extraction 38
5.2.18 EPA Method 3535A (SW-846): Solid-Phase Extraction 40
5.2.19 EPA Method 3541 (SW-846): Automated Soxhlet Extraction 43
5.2.20 EPA Method 3545A (SW-846): Pressurized Fluid Extraction (PFE) 44
5.2.21 EPA Method 3571 (SW-846): Extraction of Solid and Aqueous Samples for Chemical
Agents 47
5.2.22 EPA Method 3580A (SW-846): Waste Dilution 47
5.2.23 EPA Method 3585 (SW-846): Waste Dilution for Volatile Organics 49
5.2.24 EPA Method 5030C (SW-846): Purge-and-Trap for Aqueous Samples 50
5.2.25 EPA Method 5035A (SW-846): Closed-System Purge-and-Trap and Extraction for Volatile
Organics in Soil and Waste Samples 51
5.2.26 EPA Method 6010C (SW-846): Inductively Coupled Plasma - Atomic Emission
Spectrometry 52
5.2.27 EPA Method 6020A (SW-846): Inductively Coupled Plasma - Mass Spectrometry 53
5.2.28 EPA Method 7010 (SW-846): Graphite Furnace Atomic Absorption Spectrophotometry... 54
5.2.29 EPA Method 7470A (SW-846): Mercury in Liquid Wastes (Manual Cold-Vapor Technique)
54
5.2.30 EPA Method 747IB (SW-846): Mercury in Solid or Semisolid Wastes (Manual Cold-Vapor
Technique) 54
5.2.31 EPA Method 7473 (SW-846): Mercury in Solids and Solutions by Thermal Decomposition,
Amalgamation, and Atomic Absorption Spectrophotometry 55
5.2.32 EPA Method 7580 (SW-846): White Phosphorus by Solvent Extraction and Gas
Chromatography (GC) 55
5.2.33 EPA Method 8015C (SW-846): Nonhalogenated Organics Using GC-FID 56
5.2.34 EPA Method 8260C (SW-846): Volatile Organic Compounds by Gas Chromatography-
Mass Spectrometry (GC-MS) 56
5.2.35 EPA Method 8270D (SW-846): Semivolatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC-MS) 57
5.2.36 EPA Method 8315A (SW-846): Determination of Carbonyl Compounds by High
Performance Liquid Chromatography (HPLC) 60
5.2.37 EPA Method 8316 (SW-846): Acrylamide, Acrylonitrile and Acrolein by High Performance
Liquid Chromatography (HPLC) 60
5.2.38 EPA Method 8318A (SW-846): N-Methylcarbamates by High Performance Liquid
Chromatography (HPLC) 61
5.2.39 EPA Method 832IB (SW-846): Solvent-Extractable Nonvolatile Compounds by High
Performance Liquid Chromatography-Thermospray-Mass Spectrometry (HPLC-TS-MS) or
Ultraviolet (UV) Detection 61
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Table of Contents
5.2.40 EPA Method 8330B (SW-846): Nitroaromatics and Nitramines by High Performance
Liquid Chromatography (HPLC) 62
5.2.41 EPA ILM05.3 Cyanide: Analytical Methods for Total Cyanide Analysis 63
5.2.42 IO [Inorganic] Compendium Method IO-3.1: Selection, Preparation, and Extraction of Filter
Material 64
5.2.43 IO [Inorganic] Compendium Method IO-3.4: Determination of Metals in Ambient
Particulate Matter Using Inductively Coupled Plasma (ICP) Spectroscopy 65
5.2.44 IO [Inorganic] Compendium Method IO-3.5: Determination of Metals in Ambient
Particulate Matter Using Inductively Coupled Plasma/Mass Spectrometry (ICP-MS) 66
5.2.45 IO [Inorganic] Compendium Method IO-5: Sampling and Analysis for Vapor and Particle
Phase Mercury in Ambient Air Utilizing Cold Vapor Atomic Fluorescence Spectrometry
(CVAFS) 67
5.2.46 EPA Air Method, Toxic Organics - 10A (TO-10A): Determination of Pesticides and
Polychlorinated Biphenyls in Ambient Air Using Low Volume Polyurethane Foam (PUF)
Sampling Followed by Gas Chromatographic/Multi-Detector Detection (GC- MD) 67
5.2.47 EPA Air Method, Toxic Organics - 15 (TO-15): Determination of Volatile Organic
Compounds (VOCs) in Air Collected in Specially-Prepared Canisters and Analyzed by Gas
Chromatography/Mass Spectrometry (GC-MS) 70
5.2.48 NIOSHMethod 1612: Propylene Oxide 71
5.2.49 NIOSHMethod 2016: Formaldehyde 71
5.2.50 NIOSHMethod 2513: Ethylene Chlorohydrin 71
5.2.51 NIOSHMethod 3510: Monomethylhydrazine 72
5.2.52 NIOSH Method 5600: Organophosphorus Pesticides 72
5.2.53 NIOSH Method 5601: Organonitrogen Pesticides 73
5.2.54 NIOSH Method 6001: Arsine 73
5.2.55 NIOSH Method 6002: Phosphine 74
5.2.56 NIOSH Method 6004: Sulfur Dioxide 74
5.2.57 NIOSH Method 6010: Hydrogen Cyanide 74
5.2.58 NIOSH Method 6013: Hydrogen Sulfide 75
5.2.59 NIOSH Method 6015: Ammonia 75
5.2.60 NIOSH Method 6402: Phosphorus Trichloride 75
5.2.61 NIOSH Method 7903: Acids, Inorganic 76
5.2.62 NIOSH Method 7905: Phosphorus 76
5.2.63 NIOSH Method 7906: Fluorides, Aerosol and Gas 77
5.2.64 NIOSH Method S301-1: Fluoroacetate Anion 77
5.2.65 OSHA Method 40: Methylamine 77
5.2.66 OSHA Method 54: Methyl Isocyanate 78
5.2.67 OSHA Method 61: Phosgene 78
5.2.68 OSHA Method ID-211: Sodium Azide and Hydrazoic Acid in Workplace Atmospheres ... 79
5.2.69 OSHA Method ID-216SG: Boron Trifluoride (BF3) 79
5.2.70 OSHA Method PV2004: Acrylamide 79
5.2.71 OSHA Method PV2103: Chloropicrin 80
5.2.72 ASTM Method D5755-03: Standard Test Method for Microvacuum Sampling and Indirect
Analysis of Dust by Transmission Electron Microscopy (TEM) for Asbestos Structure
Number Surface Loading 80
5.2.73 ASTM Method D6480-99: Standard Test Method for Wipe Sampling of Surfaces, Indirect
Preparation, and Analysis for Asbestos Structure Number Concentration by Transmission
Electron Microscopy 81
5.2.74 ISO Method 10312:1995: Ambient Air - Determination of Asbestos Fibres - Direct-transfer
Transmission Electron Microscopy Method (TEM) 81
5.2.75 Standard Method 45 00-NH3 B: Nitrogen (Ammonia) Preliminary Distillation Step 82
5.2.76 Standard Method 45 00-NH3 G: Nitrogen (Ammonia) Automated Phenate Method 82
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5.2.77 Standard Method 4500-C1 G: DPD Colorimetric Method 82
5.2.78 Literature Reference for Chlorine (Analyst, 1999. 124: 1853-1857) 83
5.2.79 Literature Reference for Fluoroacetate salts (Analytical Letters, 1994. 27 (14): 2703-2718)
83
5.2.80 Literature Reference for Perfluoroisobutylene (Journal of Chromatography A, 2005. 1098:
156-165) 84
5.2.81 Literature Reference for Sodium Azide (Journal of Forensic Science, 1998. 43(1):200-202)
84
Section 6.0: Selected Radiochemical Methods 85
6.1 General Guidance 86
6.1.1 Standard Operating Procedures for Identifying Radiochemical Methods 86
6.1.2 General Quality Control (QC) Guidance for Radiochemical Methods 89
6.1.3 Safety and Waste Management 89
6.2 Method Summaries 90
6.2.1 EPA Method 111: Determination of Polonium-210 Emissions from Stationary Sources ... 91
6.2.2 EPA Method 900.0: Gross Alpha and Gross Beta Radioactivity in Drinking Water 91
6.2.3 EPA Method 901.1: Gamma Emitting Radionuclides in Drinking Water 92
6.2.4 EPA Method 903.0: Alpha-Emitting Radium Isotopes in Drinking Water 92
6.2.5 EPA Method 903.1: Radium-226 in Drinking Water - Radon Emanation Technique 93
6.2.6 EPA Method 908.0: Uranium in Drinking Water - Radiochemical Method 93
6.2.7 EPA Method EMSL-19: Determination of Radium-226 and Radium-228 in Water, Soil, Air
and Biological Tissue 94
6.2.8 EPA Method EMSL-33: Isotopic Determination of Plutonium, Uranium, and Thorium in
Water, Soil, Air, and Biological Tissue 94
6.2.9 EML HASL-300 Method Am-01-RC: Americium in Soil 95
6.2.10 EML HASL-300 Method Am-02-RC: Americium-241 in Soil-Gamma Spectrometry 96
6.2.11 EML HASL-300 Method Am-04-RC: Americium in QAP Water and Air Filters - Eichrom's
TRU Resin 96
6.2.12 EML HASL-300 Method Ga-01-R: Gamma Radioassay 97
6.2.13 EML HASL-300 Method Po-02-RC: Polonium in Water, Vegetation, Soil, and Air Filters97
6.2.14 EML HASL-300 Method Pu-12-RC: Plutonium and/or Americium in Soil or Sediments... 98
6.2.15 EML HASL-300 Method Sr-03-RC: Strontium-90 in Environmental Samples 98
6.2.16 FRMAC Method Volume 2, Page 33: Gross Alpha and Beta in Air 99
6.2.17 ORISE Method AP-1: Gross Alpha and Beta for Various Matrices 99
6.2.18 ORISE Method AP-11: Sequential Determination of the Actinides in Environmental
Samples Using Total Sample Dissolution and Extraction 100
6.2.19 ASTM Method D3084: Standard Practice for Alpha Spectrometry in Water 101
6.2.20 ASTM Method D3972: Standard Test Method for Isotopic Uranium in Water by
Radiochemistry 101
6.2.21 Standard Method 7110 B: Gross Alpha and Gross Beta Radioactivity (Total, Suspended,
and Dissolved) 102
6.2.22 Standard Method 7120: Gamma-Emitting Radionuclides 103
6.2.23 Standard Method 7500-Ra B: Radium: Precipitation Method 103
6.2.24 Standard Method 7500-Ra C: Radium: Emanation Method 104
6.2.25 Standard Method 7500-Sr B: Total Radioactive Strontium and Strontium-90: Precipitation
Method 104
6.2.26 Standard Method 7500-U B: Uranium: Radiochemical Method 105
6.2.27 Standard Method 7500-U C: Uranium: Isotopic Method 105
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Section 7.0: Selected Pathogen Methods 107
7.1 General Guidance 108
7.1.1 Standard Operating Procedures for Identifying Pathogen Methods 108
7.1.2 General Quality Control (QC) Guidance for Pathogen Methods Ill
7.1.3 Safety and Waste Management 112
7.2 Method Summaries 113
7.2.1 Laboratory Response Network (LRN) 113
7.2.2 USEPA Manual of Methods for Virology, EPA/600/4-84/013, April 2001 115
7.2.3 USEPA Environmental Regulations and Technology, Control of Pathogens and Vector
Attraction in Sewage Sludge EPA/625/R-92/013, July 2003: Baylisascaris procyonis 115
7.2.4 EPA Method 1622: Cryptosporidium in Water by Filtration/IMS/FA 116
7.2.5 EPA Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA 116
7.2.6 EPA Method 1682: Salmonella spp 117
7.2.7 U.S. FDA Bacteriological Analytical Manual, Chapter 10, 2003: Listeria monocytogenesllS
7.2.8 Standard Methods 9213 B: Staphylococcus aureus 118
7.2.9 Standard Methods 9260 B: General Qualitative Isolation and Identification Procedures for
Salmonella 119
7.2.10 Standard Methods 9260 E: Shigella 119
7.2.11 Standard Methods 9260 F: Pathogenic Escherichia coli 120
7.2.12 Standard Methods 9260 G: Campylobacter jejuni 120
7.2.13 Standard Methods 9260 H: Vibrio cholerae 121
7.2.14 Standard Methods 9260 I: Leptospira 121
7.2.15 Literature Reference for Campylobacter jejuni (Molecular and Cellular Probes. 2006. 20:
269-279) 122
7.2.16 Literature Reference for Chlamydophilapsittaci (Journal of Clinical Microbiology. 2000.
38: 1085-1093) 122
7.2.17 Literature Reference for Adenoviruses (Applied and Environmental Microbiology. 2005.
71(6): 3131-3136) 123
7.2.18 Literature Reference for Astroviruses (Canadian Journal of Microbiology. 2004. 50: 269-
278) 124
7.2.19 Literature Reference for Noroviruses (Journal of Clinical Microbiology. 2004. 42(10):
4679-4685) 124
7.2.20 Literature Reference for Sapoviruses (Journal of Medical Virology. 2006. 78(10): 1347-
1353) 125
7.2.21 Literature Reference for Coronaviruses (SARS) (Journal of Virological Methods. 2004.
122:29-36) 126
7.2.22 Literature Reference for Hepatitis E Virus (Journal of Virological Methods. 2006. 131(1):
65-71) 126
7.2.23 Literature Reference for Influenza H5N1 (Emerging Infectious Diseases. 2005. 11(8):
1303-1305) 127
7.2.24 Literature Reference for Enteric Viruses (Applied and Environmental Microbiology. 2003.
69(6): 3158-3164) 128
7.2.25 Literature Reference for Cryptosporidium spp. (Applied and Environmental Microbiology.
1999. 65(9): 3936-3941) 128
7.2.26 Literature Reference for Entamoeba histolytica (Journal of Parasitology. 1972. 58(2): 306-
310) 129
7.2.27 Literature Reference for Entamoeba histolytica (Journal of Clinical Microbiology. 2005.
43(11): 5491-5497) 129
7.2.28 Literature Reference for Giardia spp. (Transactions of the Royal Society of Tropical
Medicine and Hygiene. 1983. 77(4): 487-488) 130
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7.2.29 Literature Reference for Toxoplasma gondii (Emerging Infectious Diseases. 2006. 12(2):
326-329) 130
7.2.30 Literature Reference for Toxoplasma gondii (Applied and Environmental Microbiology.
2004. 70(7): 4035-4039) 131
Section 8.0: Selected Biotoxin Methods 133
8.1 General Guidance 134
8.1.1 Standard Operating Procedures for Identifying Biotoxin Methods 134
8.1.2 General Quality Control (QC) Guidance for Biotoxin Methods 137
8.1.3 Safety and Waste Management 138
8.2 Method Summaries 139
8.2.1 Laboratory Response Network (LRN) 139
8.2.2 U.S. FDA, Bacteriological Analytical Manual Online, Chapter 17, 2001: Botulinum
Neurotoxins 140
8.2.3 U.S. FDA, Bacteriological Analytical Manual Online, Appendix 1, 2001: Rapid Methods
for Detecting Foodborne Pathogens 141
8.2.4 AOAC Official Method 991.31: Aflatoxins in Corn, Raw Peanuts, and Peanut Butter 141
8.2.5 AOAC Official Method 993.06: Staphylococcal Enterotoxins in Selected Foods 142
8.2.6 AOAC Official Method 994.08: Aflatoxin in Corn, Almonds, Brazil Nuts, Peanuts, and
Pistachio Nuts 142
8.2.7 Literature Reference for Abrin (119th AOAC Annual Meeting & Exposition, 2005, p. 613)
143
8.2.8 Literature Reference for Abrin and Shiga and Shiga-like Toxins (Pharmacological
Toxicology. 2001. 88(5): 255-260) 143
8.2.9 Literature Reference for Abrin and Ricin (Analytical Biochemistry. 2006. 357(2): 200-207)
144
8.2.10 Literature Reference for a-Conotoxin (Biochemical Journal. 1997. 328: 245-250) 144
8.2.11 Literature Reference for a-Conotoxin (Journal of Medicinal Chemistry. 2004. 47(5): 1234-
1241) 145
8.2.12 Literature Reference for a-Amanitin, Ricin, T-2 Mycotoxin (Journal of Food Protection.
2005.68(6): 1294-1301) 145
8.2.13 Literature Reference for a-Amanitin (Journal of Chromatography. 1991. 563(2): 299-311)
146
8.2.14 Literature Reference for Anatoxin-a (Biomedical Chromatography. 1996. 10: 46-47) 146
8.2.15 Literature Reference for Brevetoxins (Environmental Health Perspectives. 2002. 110(2):
179-185) 147
8.2.16 Literature Reference for Brevetoxins (Toxicon. 2004. 43(4): 455-465) 147
8.2.17 Literature Reference for Cylindrospermopsin (FEMS Microbiology Letters. 2002. 216(2):
159-164) 148
8.2.18 Literature Reference for Diacetoxyscirpenol (DAS) (International Journal of Food
Microbiology. 1988. 6(1): 9-17) 148
8.2.19 Literature Reference for Diacetoxyscirpenol (DAS) and T-2 Mycotoxin (Rapid
Communications in Mass Spectrometry. 2006. 20(9): 1422-1428) 149
8.2.20 Literature Reference for Microcystins (Journal of AOAC International. 2001. 84(4): 1035-
1044) 149
8.2.21 Literature Reference for Microcystins (Analyst. 1994. 119(7): 1525-1530) 150
8.2.22 Literature Reference for Picrotoxin (Journal of Pharmaceutical & Biomedical Analysis.
1989. 7(3): 369-375) 150
8.2.23 Literature Reference for Saxitoxin (Journal of AOAC International. 1995. 78(2): 528-532)
151
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8.2.24 Literature Reference for Shiga and Shiga-like Toxin (Journal of Clinical Microbiology.
2007. 45(10): 3377-3380) 151
8.2.25 Literature Reference for Tetrodotoxin (Journal of Clinical Laboratory Analysis. 1992. 6(2):
65-72) 152
8.2.26 Literature Reference for Tetrodotoxin (Analytical Biochemistry. 2001. 290(1): 10-17)... 152
8.2.27 Lateral Flow Immunoassay Kits 153
Section 9.0: Conclusions 155
Appendices
Appendix A: Selected Chemical Methods A-l
Appendix B: Selected Radiochemical Methods B-l
Appendix C: Selected Pathogen Methods C-l
Appendix D: Selected Biotoxin Methods D-l
Figures
Figure 1-1. Environmental Evaluation Analytical Process Roadmap for Homeland Security Events 2
Figure 2-1. SAM Method Selection Process 5
Tables
Table 5-1. Chemical Methods and Corresponding Text Section Numbers 14
Table 5-2. Sources of Chemical Methods 27
Table 6-1. Radiochemical Methods and Corresponding Text Section Numbers 86
Table 6-2. Sources of Radiochemical Methods 88
Table 7-1. Pathogen Methods and Corresponding Text Section Numbers 109
Table 7-2. Sources of Pathogen Methods 110
Table 8-1. Biotoxin Methods and Corresponding Text Section Numbers 135
Table 8-2. Sources of Biotoxin Methods 136
SAM Revision 3.1 xxiii November 15, 2007
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SAM Revision 3.1 xxiv November 15, 2007
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Section 1 - Introduction
Section 1.0: Introduction
In the aftermath of the terrorist attacks of September 11, 2001, and the anthrax attacks in the fall of 2001,
federal and state personnel carried out their mission to provide response, recovery, and remediation under
trying circumstances, including an unprecedented demand on their capabilities to analyze environmental
samples. In reviewing these events, EPA identified several areas where the country could better prepare
itself in the event of future terrorist incidents. One of the most important areas identified was the need to
improve the nation's laboratory capacity and capability to analyze environmental samples following a
homeland security event.
In response, EPA formed the Homeland Security Laboratory Capacity Workgroup to identify and
implement opportunities for near-term improvements and to develop recommendations for addressing
longer-term laboratory issues. The EPA Homeland Security Laboratory Capacity Workgroup consists of
representatives from the Office of Research and Development, Office of Air and Radiation, Office of
Water, Office of Solid Waste and Emergency Response, Office of Environmental Information, Office of
Pollution Prevention and Toxics, and several EPA regional offices.
A critical area identified by the workgroup was the need for a list of analytical methods to be used by all
laboratories when analyzing homeland security event samples and, in particular, when analysis of a large
number of samples is required over a short period of time. Having standardized methods would reduce
confusion, permit sharing of sample load between laboratories, improve data comparability, simplify the
task of outsourcing analytical support to the commercial laboratory sector, and improve the follow-up
activities of validating results, evaluating data, and making decisions. To this end, workgroup members
formed an Analytical Methods Subteam to address homeland security methods issues.
The Analytical Methods Subteam recognized that widely different analytical methods are required for
various phases of environmental sample analyses in support of homeland security preparation and
response: (1) ongoing surveillance and monitoring, (2) response and rapid screening for determining
whether an event has occurred, (3) preliminary site characterizations to determine the extent and type of
contamination, and (4) confirmatory laboratory analyses to plan, implement, and evaluate the
effectiveness of site remediation. Figure 1-1 represents these analytical phases. EPA's Standardized
Analytical Methods for Environmental Restoration following Homeland Security Events (SAM) provides
information for analytical methods to be applied during the "Site Remediation" phase.
SAM Revision 3.1 1 November 15, 2007
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Section 1 - Introduction
Figure 1-1. Environmental Evaluation Analytical Process Roadmap for Homeland
Security Events
Surveillance and Monitoring
Immediate Response/
Credibility Determination
Preliminary Site
Characterization
SAM
Site Remediation
• Assessment
»Cleanup
• Clearance
SAM
(Standardized Analytical Methods for Environmental
Restoration following Homeland Security Events)
SAM Revision 3.1
November 15, 2007
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Section 2 - Background
Section 2.0: Background
SAM identifies a single method per analyte/sample type to ensure a consistent analytical approach across
multiple laboratories when analyzing environmental samples following an event. In support of this
document, EPA periodically assembles methods experts from within EPA and other federal agencies to
review methods and, if necessary, revise the methods listed. Method selection is based on consideration
of specific criteria that emphasize method performance and include existing laboratory capabilities,
laboratory capacity, method applicability to multiple environmental sample types, and method
applicability to multiple SAM analytes. For some analytes, the preferred method is a clear choice; for
others, competing criteria make the choice more difficult. Final method selections are based on technical
recommendations from the SAM work groups. For analytes where limited laboratory testing/experience
exists, such as chemical warfare agents, methods were selected based on their applicability to similar
chemicals (e.g., nerve agents and some pesticides). In these cases, laboratory studies to test the ability of
the selected method to measure the target analyte(s) are planned. Figure 2-1 summarizes steps and
provides the criteria used during the SAM method selection process. It is important to note that the
method selection criteria included in this figure are listed in non-hierarchical order and, in some cases,
only a subset of the criteria was considered.
In 2004, EPA's National Homeland Security Research Center (NHSRC) brought together experts from
across EPA and its sister agencies to develop a compendium of analytical methods to be used when
analyzing environmental samples to address site characterization, remediation and clearance following
future homeland security events. Participants included representatives from EPA program offices, EPA
regions, EPA national laboratories, Centers for Disease Control and Prevention (CDC), Food and Drug
Administration (FDA), Department of Homeland Security (DHS), Federal Bureau of Investigation (FBI),
Department of Defense (DoD), Department of Agriculture (USDA), and U.S. Geological Survey (USGS).
Methodologies were considered for chemical and biological agents of concern in the types of
environmental samples that would be anticipated. The primary objective of this effort was to identify
appropriate SAM Analytical Methods Subteam consensus methods that represent a balance between
providing existing, documented, determinative techniques and providing consistent and valid analytical
results.
A survey of available confirmatory analytical methods for approximately 120 biological and chemical
analytes was conducted using existing resources including the following:
• National Environmental Methods Index (NEMI) and NEMI-Chemical, Biological, and Radiological
(NEMI-CBR)
• Environmental Monitoring Method Index (EMMI)
• EPA Test Methods Index
EPA Office of Solid Waste SW-846 Methods
• EPA Microbiology Methods
• National Institute for Occupational Safety and Health (NIOSH) method index
• Occupational Safety and Health Administration (OSHA) method index
• AOAC International
• ASTM International
• International Organization for Standardization (ISO) methods
• Standard Methods for the Examination of Water and Wastewater
• PubMED Literature Database
In September 2004, EPA published Standardized Analytical Methods for Use During Homeland Security
Events, Revision 1.0 (EPA/600/R-04/126), SAM Revision 1.0, which provided a list of analytical and
sample preparation methods that were selected for measurement of 82 chemical analytes in
aqueous/liquid, solid, oily solid, and air samples, and 27 biological analytes in water, dust, and aerosol
SAM Revision 3.1 3 November 15, 2007
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Section 2 - Background
samples. During 2005, SAM was expanded to include radioisotopes, several persistent chemical warfare
agent degradation products, a drinking water sample type, methods for determination of the viability of
biological organisms, and a separate section for biotoxin analytes. Where necessary, the methods
included in SAM Revision 1.0 were updated to reflect more recent or appropriate methodologies. Similar
efforts to those used for method selection during development of SAM Revision 1.0 were undertaken to
select and include methods for measurement of radioisotopes and chemical warfare agent degradation
products in all sample types, for measurement of chemical, biological, and radiochemical analytes in
drinking water, and to determine the viability of biological organisms. These additional analytes and the
corresponding methods selected were included in SAM Revision 2.0.
During 2006, SAM was revised further to incorporate analytes included on updated federal agency lists,
provide additional or more current method listings for target analytes, incorporate explosives into the
chemical analytes listing, combine identification and viability methods information for pathogens, and
address comments from EPA Science Advisory Board's Homeland Security Advisory Committee1 to
clarify the intended use of the document. These changes were included in SAM Revision 3.0. Following
publication of SAM Revision 3.0, NHSRC plans to convene SAM Technical Workgroups at least once
per year, to evaluate and, if necessary, update the analytes and methods that are listed in SAM. This
version, SAM Revision 3.1, reflects the first of these updates and includes the addition of several
chemical analytes, one radionuclide, and one biotoxin, along with corresponding selected methods.
NHSRC plans to provide the updated information in a Web-based version of the document that will allow
users and other stakeholders to search for specific needs and to submit questions and comments regarding
the information.
JEPA Science Advisory Board's Homeland Security Advisory Committee meeting summary:
hitp://www.cpa.gov/sab/06minuics/lisac ws and sain 01 30-31 06 minntcs.pdf
SAM Revision 3.1 4 November 15, 2007
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Section 2 - Background
Figure 2-1. SAM Method Selection Process
Step 1
Is there an EPA
published method f
analyte in the matrix of
5 there a method that has beei
developed and published by another
Standard Body (VCSB) for measun
of the analyte in the matrix of interest
s there an EPA, federal, o
VCSB method that has been
of the analyte in another
jrement of the analyte
n the matrix of interest
Evaluate method against
selection criteria
NO
Repeat Steps 1 - 4 to identify methods that measure
analytes s/m//arto the analyte of concern
Use the following criteria as guidelines to assess which method is most
appropriate for inclusion in SAM:
• Has the method been tested/approved by issuing program office?
• Has the method been evaluated based on reliability, performance criteria
(e.g., sensitivity, specificity, false positives/false negatives, precision,
recovery)?
• Is the method appropriate for measurement of this analyte in the matrix
of interest to assess extent of contamination and decontamination
effectiveness?
• Has the method been tested for the specific intended use?
• Is the existing lab capacity (i.e., equipment, number of labs, cost) suitable
for implementation of the method?
• Is the required equipment readily available?
• Is the method capable of determining viability of an organism?
• What is the time required for analysis?
• Are reagents, standards, controls, etc., available and accessible?
• Is specific and/or unique training required?
• Are large sample volumes required?
• Are analytical costs high?
• Has the method already been selected for other SAM analytes?
• Are modifications needed to accommodate the analytes or matrices?
Select meth<
lod for inclusion in SAM j
I If no methods are available, prioritize
for further research
SAM Revision 3.1
November 15, 2007
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Section 2 - Background
SAM Revision 3.1 6 November 15, 2007
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Section 3 - Scope and Application
Section 3.0: Scope and Application
The premise and purpose of this document is to standardize the analytical methods that will be used in
cases when multiple laboratories are called on to analyze environmental samples following a homeland
security event. The document also is intended as a tool that will be available to assist state and local
laboratories in planning for and analyzing environmental samples following a homeland security event.
The methods presented in this document should be used to:
• Confirm the identification of contaminants;
Evaluate the extent of contamination; and
Evaluate the effectiveness of decontamination.
The list of methods provided is limited to those methods that would be used to determine, to the extent
possible within analytical limitations, the presence of chemical, radiochemical, pathogen, and biotoxin
analytes of concern and their concentrations in environmental media. The methods include detailed
laboratory procedures for confirming the identification of analytes and determining their concentrations in
environmental samples. The methods, therefore, are not designed to be used for rapid or immediate
response or for conducting an initial evaluation (triage or screening) of suspected material to determine if
it poses an immediate danger or should be analyzed in specially designed, highly secure facilities. This
document also is not intended to provide information regarding sample collection activities or equipment.
Methods for addressing these needs are and will be the subject of other efforts.
Methods are provided in this document as corresponding to specific analyte/sample type combinations
that are listed in Appendices A (chemical), B (radiochemical), C (pathogen), and D (biotoxin).
Summaries of each method are provided in numerical order by the developing agency, throughout
Sections 5.2 (chemical methods), 6.2 (radiochemical methods), 7.2 (pathogen methods), and 8.2 (biotoxin
methods).
It is important to note that, in some cases, the methods included in this document have not been fully
validated for the analyte/sample type combination(s) for which they have been selected. The
information contained in this document represents the latest step in an ongoing National Homeland
Security Research Center effort to provide standardized analytical methods for use by laboratories
(e.g., EPA contract laboratories) tasked with performing confirmatory analyses on environmental
samples following a homeland security incident. Although at this time, some of the methods listed
have not been fully validated for a particular analyte (e.g., analytes not explicitly identified in the
method) or sample type, the methods are considered to contain the most appropriate currently
available techniques. Unless a published method that is listed in this document states specific
applicability to the analyte/sample type combination for which it has been selected, it should be
assumed that method testing is needed, and adjustments may be required to accurately account for
variations in analyte characteristics, environmental samples, and target risk levels. Many of the
target analytes listed in this document have only recently become an environmental concern. EPA is
actively pursuing development and validation of Standard Analytical Protocols (SAPs) based on the
methods listed, including optimization of procedures for measuring target compounds. In those cases
where method procedures are determined to be insufficient for a particular situation, EPA will
provide guidance regarding appropriate actions. This will be an ongoing process as EPA will strive
to establish a consistent level of validation for all listed analytes.
SAM Revision 3.1 7 November 15, 2007
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Section 3 - Scope and Application
EPA recognizes that specification of a single method may limit laboratory capacity and techniques that
may be needed to evaluate difficult samples. In those cases where method procedures are determined to
be insufficient for a particular situation, EPA will provide guidance regarding appropriate actions (see list
of contacts in Section 4). EPA is developing and validating Standardized Analytical Protocols (SAPs)
based on the methods that are listed in this document, where further development and testing are
necessary. Once validation is complete, data regarding the resulting method performance and data quality
objectives will be available. The SAM document and corresponding SAPs will be reviewed frequently.
EPA plans to continue to update the SAM document to address the needs of homeland security, reflect
improvements in analytical methodology and new technologies, and incorporate changes in analytes
based on needs. EPA also anticipates that addenda may be generated to provide guidance regarding
issues that currently are not addressed by this document. Any deviations from the methods referenced in
this document should be coordinated with the appropriate point(s) of contact identified in Section 4.
Participants in the chemical, radiochemical, pathogen, and biotoxin work groups, including
representatives from the U.S. EPA, CDC, FDA, DHS, FBI, DoD, USDA, and USGS evaluated the
suitability of existing methodologies and selected this set of methods for use by EPA laboratories and
contract laboratories if called upon in an emergency. EPA recognizes that this advanced selection of such
methods may pose potential risks, including the following:
• Selecting technologies that may not be the most cost-effective technologies currently available for
addressing the particular situation at hand;
• Selecting methodologies that may not be appropriate for use in responding to a particular
emergency because EPA did not anticipate having to analyze for a particular analyte or
analyte/sample type combination; and
Preventing development and adoption of new and better measurement technologies.
To address these potential risks as soon as possible, EPA plans to take several steps. These include the
following:
Developing and specifying measurement quality objectives (i.e., required minimum standards of
accuracy (bias and precision) and sensitivity for the analysis of samples that support the data
quality needs of the particular stage of the emergency response/recovery process) for all
analyte/sample type combinations listed in this document;
Specifying guidance for ensuring the analytical methods listed provide results that are consistent
with and support their intended use;
Working with other government agencies and the private sector to establish a laboratory network to
ensure that laboratories selected to assist EPA and its federal, state, and local partners in responding
to homeland security events have the requisite expertise and systems to perform this type of testing;
and
Continuing to work with multiple agencies and stakeholders to update SAM and supporting
documents periodically.
EPA recognizes that having data of known and documented quality is critical for public officials to assess
accurately the activities that may be needed in remediating a site during and following emergency
situations. Data must be of sufficient quality to support decision making. Quality control (QC), however,
takes time, and time is often critical in emergency-related activities where there will be tremendous
pressure to conduct sampling and analytical operations quickly and efficiently. While reduced levels of
QC might be tolerated during the rapid screening stage of emergency response, implementation of
analytical methods for risk assessment and site release will require a higher and more appropriate level of
QC. Many of the methods listed in this document include QC requirements for collecting and analyzing
samples. These QC requirements may or may not be appropriate for addressing emergency response
SAM Revision 3.1 8 November 15, 2007
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Section 3 - Scope and Application
situations, and may be adjusted as necessary to maximize data and decision quality. Specific QC
recommendations for analysis of samples for chemical, radiochemical, pathogen, and biotoxin analytes
are provided in each corresponding section of this document (i.e., Sections 5.1.2, 6.1.2, 7.1.2, and 8.1.2,
respectively).
SAM Revision 3.1 9 November 15, 2007
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Section 3 - Scope and Application
SAM Revision 3.1 10 November 15, 2007
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Section 4 - Points of Contact
Section 4.0: Points of Contact
Questions concerning this document, or the methods identified in this document, should be addressed to
the appropriate point(s) of contact identified below. These contacts should be consulted regarding any
method deviations or modifications, sample problems or interferences, quality control requirements, or
the use of potential alternative methods. As previously indicated, any deviations from the recommended
method(s) should be reported immediately to ensure data comparability is maintained when responding to
homeland security events.
General
Oba Vincent - Primary
National Homeland Security Research Center
U.S. EPA Office of Research and Development (NG16)
26 West Martin Luther King Jr. Drive
Cincinnati, OH 45268
(513)569-7456
Rob Rothman - Alternate
National Homeland Security Research Center
U.S. EPA Office of Research and Development (NG16)
26 West Martin Luther King Jr. Drive
Cincinnati, OH 45268
(513)569-7187
Chemical Methods
Barry Lesnik - Primary
Steve Reimer - Alternate
Office of Solid Waste and Emergency Response (5307P) USEPA Region 10 - Manchester Laboratory
U.S. EPA Headquarters Ariel Rios Building
1200 Pennsylvania Avenue, NW
Washington, DC 20460
(703)308-0476
7411 Beach Drive East
Port Orchard, WA 98366
(360)871-8718
re i me r. ste ve@e pa. gov
Radiochemical Methods
John Griggs - Primary
U.S. EPA Office of Radiation and Indoor Air
Environmental Laboratory
540 South Morris Avenue
Montgomery, AL 36115-2601
(334) 270-3450
Kathy Hall - Alternate
National Homeland Security Research Center
U.S. EPA Office of Research and Development (NG16)
26 West Martin Luther King Jr. Drive
Cincinnati, OH 45268
(513)379-5260
haU.kafliYMcBa.gQY
Pathogen Methods
Alan Lindquist - Primary
National Homeland Security Research Center
U.S. EPA Office of Research and Development (NG16)
26 West Martin Luther King Jr. Drive
Cincinnati, OH 45268
(513)569-7192
Sanjiv Shah -Alternate
National Homeland Security Research Center
U.S. EPA Office of Research and Development (NG16)
26 West Martin Luther King Jr. Drive
Cincinnati, OH 45268
(513)569-7776
Biotoxins Methods
Matthew Magnuson
National Homeland Security Research Center
U.S. EPA Office of Research and Development (NG16)
26 West Martin Luther King Jr. Drive
Cincinnati, OH 45268
(513)569-7321
magnuson.matthcw@cpa.gov
Sanjiv Shah -Alternate
National Homeland Security Research Center
U.S. EPA Office of Research and Development (NG16)
26 West Martin Luther King Jr. Drive
Cincinnati, OH 45268
(513)569-7776
iv@epa.gov
SAM Revision 3.1
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November 15, 2007
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Section 4 - Points of Contact
SAM Revision 3.1 12 November 15, 2007
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Section 5 - Selected Chemical Methods
Section 5.0: Selected Chemical Methods
A list of methods to be used in analyzing environmental samples for chemical contaminants during
remediation activities following a homeland security event is provided in Appendix A. Methods are listed
for each analyte and for each sample type that potentially may need to be measured and analyzed when
responding to an environmental emergency. Procedures from peer-reviewed journal articles are listed for
those analyte-sample type combinations where methods are not available. Once standard procedures are
available, the literature references will be replaced.
Please note: This section provides guidance for selecting chemical methods that have a high likelihood
of assuring analytical consistency when laboratories are faced with a large scale environmental
restoration crisis. Not all methods have been verified for the analyte/sample type combination listed in
Appendix A. Please refer to the specified method to identify analyte/sample type combinations that
have been verified. Any questions regarding information discussed in this section should be addressed
to the appropriate contact(s) listed in Section 4.
Appendix A is sorted alphabetically by analyte and includes the following information:
Analyte(s). The component, contaminant, or constituent of interest.
• Chemical Abstract Survey Registration Number (CAS RN). A unique identifier for chemical
substances that provides an unambiguous way to identify a chemical or molecular structure when
there are many possible systematic, generic, or trivial names.
Determinative technique. An analytical instrument or technique used to determine the quantity and
identification of compounds or components in a sample.
Method type. Two method types (sample preparation and determinative) are used to complete
sample analysis. In some cases, a single method contains information for both sample preparation
and determinative procedures. In most instances, however, two separate methods may need to be
used in conjunction.
Solid samples. The recommended method/procedure to identify and measure the analyte of interest
in solid phase samples.
• Non-aqueous liquid/organic solid samples. The recommended method/procedure to identify and
measure the analyte of interest in non-aqueous liquid/organic phase samples. An organic solid
sample is a solid that completely dissolves in an organic solvent and leaves no solid residue.
Aqueous liquid samples. The recommended method/procedure to identify and measure the analyte
of interest in aqueous liquid phase samples.
Drinking water samples. The recommended method/procedure to identify and measure the analyte
of interest in drinking water samples.
Air samples. The recommended method/procedure to identify and measure the analyte of interest in
air samples.
SAM Revision 3.1 13 November 15, 2007
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Section 5 - Selected Chemical Methods
5.1 General Guidance
This section provides a general overview of how to identify the appropriate chemical method(s) for a
given analyte-sample type combination as well as recommendations for quality control procedures.
For additional information on the properties of the chemicals listed in Appendix A, TOXNET
Qittgy/toxncyilnijiih.goWindcjyibtil), a cluster of databases on toxicology, hazardous chemicals, and
related areas maintained by the National Library of Medicine, is an excellent resource. Additional
resources include:
Syracuse Research Corporation's Physprop and Chemfate, part of the Environmental Fate Database
supported by EPA. :7/www.syrrcs.com/csc/databascs.htm
• INCHEM at http://www.inchem.org/ contains both chemical and toxicity information.
• The Registry of Toxic Effects of Chemical Substances (RTECS) database can be accessed via the
National Institiute for Occupational Safety and Health (NIOSH) Web site at
http://www.cdc.gov/niosh/rtecs/vz72d288.htmltfJWIPAW for toxicity information.
• EPA's Integrated Risk Information System (IRIS): http://www.cpa.gov/iris/ contains toxicity
information.
The Forensic Science and Communications Journal published by the Laboratory Division of the
Federal Bureau of Investigation, http://www.fti.gov/liq/lab/fsc/ciirrent/backissu.htm
European Chemicals Bureau Toxicology and Chemical Substances: httg^/ecbjrcjt and
http://ccb.jrc.it/tcsting-mctliods/ containing information regarding European Directive 67/548/EEC
and Annex V
Additional research on chemical contaminants is ongoing within EPA, and databases to manage this
information are currently under development.
5.1.1 Standard Operating Procedures for Identifying Chemical Methods
To determine the appropriate method that is to be used on an environmental sample, locate the analyte of
concern in Appendix A: Chemical Methods under the "Analyte(s)" column. After locating the analyte of
concern, continue across the table to identify the appropriate determinative technique (e.g., HPLC, GC-
MS), then identify the appropriate sample preparation and determinative method(s) for the sample type of
interest (solid, non-aqueous liquid/organic solid, aqueous liquid, drinking water, or air). In some cases,
two methods (sample preparation and determinative) are needed to complete sample analysis.
Sections 5.2.1 through 5.2.81 below provide summaries of the sample preparation and determinative
methods listed in Appendix A. Once a method has been identified in Appendix A, Table 5-1 can be used
to locate the method summary.
Table 5-1. Chemical Methods and Corresponding Text Section Numbers
Analyte
Acrylamide
Acrylonitrile
CASRN
79-06-1
107-13-1
Method
8316(EPASW-846)
PV2004 (OSHA)
Section
5.2.37
5.2.70
SAM Revision 3.1 14 November 15, 2007
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Section 5 - Selected Chemical Methods
Analyte
Aldicarb (Temik)
Allyl alcohol
2-Amino-4,6-dinitrotoluene (2-Am-DNT)
4-Amino-2,6-dinitrotoluene (4-Am-DNT)
4-Aminopyridine
Ammonia
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
Arsine
Asbestos
Boron trifluoride
Brodifacoum
Bromadiolone
Carbofuran (Furadan)
Carbon disulfide
CASRN
116-06-3
107-18-6
35572-78-2
19406-51-0
504-24-5
7664-41-7
7803-55-6
7440-38-2
1327-53-3
7784-42-1
1332-21-4
7637-07-2
56073-10-0
28772-56-7
1563-66-2
75-15-0
Method
531. 2 (EPA OW)
8318A(EPASW-846)
5601 (NIOSH)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
TO-15(EPAORD)
3535A (EPA SW-846)
8330B (EPA SW-846)
350.1 (EPAOW)
6015 (NIOSH)
4500-NHs B (SM)
4500-NHs G (SM)
200.8 (EPA OW)
3031 (EPA SW-846)
3050B (EPA SW-846)
601 OC (EPA SW-846)
6020A (EPA SW-846)
IO-3.1 (EPAORD)
IO-3.4 (EPA ORD)
IO-3.5 (EPAORD)
200.8 (EPA OW)
3050B (EPA SW-846)
7010 (EPA SW-846)
6001 (NIOSH)
D5755-03 (ASTM)
D6480-99 (ASTM)
10312:1995 (ISO)
ID-216SG(OSHA)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 (EPA SW-846)
531. 2 (EPAOW)
831 8A (EPA SW-846)
5601 (NIOSH)
524.2 (EPA OW)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
TO-15(EPAORD)
Section
5.2.11
5.2.38
5.2.53
5.2.23
5.2.24
5.2.25
5.2.34
5.2.47
5.2.18
5.2.40
5.2.7
5.2.59
5.2.75
5.2.76
5.2.2
5.2.15
5.2.16
5.2.26
5.2.27
5.2.42
5.2.43
5.2.44
5.2.2
5.2.16
5.2.28
5.2.54
5.2.72
5.2.73
5.2.74
5.2.68
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.39
5.2.11
5.2.38
5.2.53
5.2.9
5.2.23
5.2.24
5.2.25
5.2.34
5.2.47
SAM Revision 3.1
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Section 5 - Selected Chemical Methods
Analyte
Chlorfenvinphos
Chlorpyrifos
Chlorine
2-Chloroethanol
3-Chloro-1 ,2-propanediol
Chloropicrin
Chlorosarin
Chlorosoman
2-Chlorovinylarsonous acid (CVAA)
Crimidine
CASRN
470-90-6
2921-88-2
7782-50-5
107-07-3
96-24-2
76-06-2
1445-76-7
7040-57-5
85090-33-1
535-89-7
Method
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
4500-CI G (SM)
Analyst, Vol. 124, 1999, pp.
1853-1857
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
2513(NIOSH)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
551 (EPA OW)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
PV2103(OSHA)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
200.8 (EPA OW)
3031 (EPA SW-846)
3050B (EPA SW-846)
601 OC (EPA SW-846)
6020A (EPA SW-846)
IO-3.1 (EPAORD)
IO-3.4 (EPA ORD)
IO-3.5 (EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
Section
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.77
5.2.78
5.2.23
5.2.24
5.2.25
5.2.34
5.2.50
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.13
5.2.20
5.2.22
5.2.35
5.2.71
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.2
5.2.15
5.2.16
5.2.26
5.2.27
5.2.42
5.2.43
5.2.44
5.2.18
5.2.20
5.2.22
5.2.35
SAM Revision 3.1
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Section 5 - Selected Chemical Methods
Analyte
Cyanide, Total
Cyanogen chloride
Cyclohexyl sarin (GF)
1,2-Dichloroethane
Dichlorvos
Dicrotophos
Diesel Range Organics
Diisopropyl methylphosphonate (DIMP)
Dimethylphosphite
CASRN
57-12-5
506-77-4
329-99-7
107-06-2
62-73-7
141-66-2
NA
1445-75-6
868-85-9
Method
335.4 (EPA OW)
ILM05.3 CN (EPA CLP)
6010(NIOSH)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
TO-15(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
524.2 (EPA OW)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
TO-15(EPAORD)
525.2 (EPA OW)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
801 5C (EPA SW-846)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
Section
5.2.6
5.2.41
5.2.57
5.2.23
5.2.24
5.2.25
5.2.34
5.2.47
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.9
5.2.23
5.2.24
5.2.25
5.2.34
5.2.47
5.2.10
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.33
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
SAM Revision 3.1
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Section 5 - Selected Chemical Methods
Analyte
Dimethylphosphoramidic acid
3,5-Dinitroaniline (3,5-DNA)
1,3-Dinitrobenzene (1,3-DNB)
2,4-Dinitrotoluene (2,4-DNT)
2,6-Dinitrotoluene (2,6-DNT)
Diphacinone
1,4-Dithiane
EA2192 [Diisopropylaminoethyl methyl-
thiophosphonate]
Ethyl methylphosphonic acid (EMPA)
Ethyldichloroarsine (ED)
N-Ethyldiethanolamine (EDEA)
Ethylene oxide
CASRN
33876-51-6
618-87-1
99-65-0
121-14-2
606-20-2
82-66-6
505-29-3
73207-98-4
1832-53-7
598-14-1
139-87-7
75-21-8
Method
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
8330B (EPA SW-846)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-15(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
TO-15(EPAORD)
Section
5.2.18
5.2.20
5.2.22
5.2.39
5.2.46
5.2.18
5.2.40
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.39
5.2.18
5.2.20
5.2.22
5.2.35
5.2.18
5.2.20
5.2.22
5.2.39
5.2.46
5.2.18
5.2.20
5.2.22
5.2.35
5.2.47
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.39
5.2.46
5.2.23
5.2.24
5.2.25
5.2.34
5.2.47
SAM Revision 3.1
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Section 5 - Selected Chemical Methods
Analyte
Fenamiphos
Fentanyl
Fluoride
Fluoroacetic acid and fluoroacetate salts
Formaldehyde
Gasoline Range Organics
Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)
Hexamethylenetriperoxidediamine (HMTD)
Hydrogen bromide
Hydrogen chloride
Hydrogen cyanide
Hydrogen fluoride
Hydrogen sulfide
Isopropyl methylphosphonic acid (IMPA)
Kerosene
CASRN
22224-92-6
437-38-7
16984-48-8
NA
50-00-0
NA
121-82-4
283-66-9
10035-10-6
7647-01-0
74-90-8
7664-39-3
7783-06-4
1832-54-8
64742-81-0
Method
525.2 (EPA OW)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
300.1, Rev 1.0 (EPA OW)
300.1, Rev 1.0 (EPA OW)
S301-1 (NIOSH)
Analytical Letters, 1994, 27
(14), 2703-2718
831 5A (EPA SW-846)
2016 (NIOSH)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
801 5C (EPA SW-846)
3535A (EPA SW-846)
8330B (EPA SW-846)
7903 (NIOSH)
335.4 (EPA OW)
ILM05.3 CN (EPA CLP)
6010 (NIOSH)
7903 (NIOSH)
6013 (NIOSH)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
801 5C (EPA SW-846)
Section
5.2.10
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.39
5.2.46
5.2.5
5.2.5
5.2.64
5.2.79
5.2.36
5.2.49
5.2.23
5.2.24
5.2.25
5.2.33
5.2.18
5.2.40
5.2.61
5.2.6
5.2.41
5.2.57
5.2.61
5.2.58
5.2.18
5.2.20
5.2.22
5.2.39
5.2.46
5.2.23
5.2.24
5.2.25
5.2.33
SAM Revision 3.1
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Section 5 - Selected Chemical Methods
Analyte
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite oxide
Mercury, Total
Methamidophos
Methyomyl
Methyl acrylonitrile
Methoxyethylmercuric acetate
Methyl fluoroacetate
Methyl hydrazine
Methyl isocyanate
Methyl parathion
Methyl-2,4,6-trinitrophenylnitramine (Tetryl)
Methylamine
CASRN
541-25-3
40334-69-8
40334-70-1
1306-02-1
7439-97-6
10265-92-6
16752-77-5
126-98-7
151-38-2
453-18-9
60-34-4
624-83-9
298-00-0
479-45-8
74-89-5
Method
200.8 (EPA OW)
3031 (EPA SW-846)
3050B (EPA SW-846)
601 OC (EPA SW-846)
6020A (EPA SW-846)
IO-3.1 (EPAORD)
IO-3.4 (EPA ORD)
IO-3.5 (EPAORD)
245.2 (EPA OW)
7473 (EPA SW-846)
IO-5 (EPA ORD)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
531. 2 (EPA OW)
831 8A (EPA SW-846)
5601 (NIOSH)
831 6 (EPA SW-846
PV2004 (OSHA)
245.2 (EPA OW)
7473 (EPA SW-846)
IO-5 (EPA ORD)
300.1, Rev 1.0 (EPA OW)
S301-1 (NIOSH)
Analytical Letters, 1994, 27
(14), 2703-2718
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
3510 (NIOSH)
OSHA 54
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
8330B (EPA SW-846)
OSHA 40
Section
5.2.2
5.2.15
5.2.16
5.2.26
5.2.27
5.2.42
5.2.43
5.2.44
5.2.3
5.2.31
5.2.45
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.11
5.2.38
5.2.53
5.2.37
5.2.70
5.2.3
5.2.31
5.2.45
5.2.5
5.2.64
5.2.79
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.51
5.2.66
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.40
5.2.65
SAM Revision 3.1
20
November 15, 2007
-------
Section 5 - Selected Chemical Methods
Analyte
N-Methyldiethanolamine (MDEA)
1-Methylethyl ester ethylphosphonofluoridic
acid (GE)
Methylphosphonic acid (MPA)
Mevinphos
Mustard, nitrogen (HN-1) [bis(2-chloroethyl)-
ethylamine]
Mustard, nitrogen (HN-2) [2,2'-dichloro-N-
methyldiethylamine N,N-bis(2-chloroethyl)-
methylamine]
Mustard, nitrogen (HN-3) [tris(2-chloroethyl)-
amine]
Mustard, sulfur (HD) / Mustard gas (H)
Nicotine sulfate
Nitrobenzene (NB)
Nitroglycerine (NG)
2-Nitrotoluene (2-NT)
3-Nitrotoluene (3-NT)
4-Nitrotoluene (4-NT)
Octahydro-1, 3,5, 7-tetranitro-1, 3,5,7-
tetrazocine (HMX)
CASRN
105-59-9
1189-87-3
993-13-5
7786-34-7
538-07-8
51-75-2
555-77-1
505-60-2
54-11-5
98-95-3
55-63-0
88-72-2
99-08-1
99-99-0
2691-41-0
Method
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
525.2 (EPA OW)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3571 (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
3535A (EPA SW-846)
8330B (EPA SW-846)
3535A (EPA SW-846)
8330B (EPA SW-846)
Section
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.39
5.2.46
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.20
5.2.22
5.2.39
5.2.46
5.2.10
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.21
5.2.35
5.2.46
5.2.18
5.2.20
5.2.22
5.2.35
5.2.18
5.2.40
5.2.18
5.2.40
SAM Revision 3.1
21
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-------
Section 5 - Selected Chemical Methods
Analyte
Organophosphate pesticides, NOS
Osmium tetroxide
Oxamyl
Paraquat
Parathion
Pentaerythritol tetranitrate (PETN)
Perfluoroisobutylene (PFIB)
Phencyclidine
Phenol
Phorate
Phosgene
CASRN
NA
20816-12-0
23135-22-0
4685-14-7
56-38-2
78-11-5
382-21-8
77-10-1
108-95-2
298-02-2
75-44-5
Method
507 (EPA OW)
61 4 (EPA OW)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
5600 (NIOSH)
252.2 (EPA OW)
3050B (EPA SW-846)
601 OC (EPA SW-846)
IO-3.1 (EPAORD)
IO-3.4 (EPA ORD)
531. 2 (EPA OW)
831 8A (EPA SW-846)
5601 (NIOSH)
549.2 (EPA OW)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
8330B (EPA SW-846)
OSHA61
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
OSHA61
Section
5.2.8
5.2.14
5.2.19
5.2.20
5.2.22
5.2.35
5.2.52
5.2.4
5.2.16
5.2.26
5.2.42
5.2.43
5.2.11
5.2.38
5.2.53
5.2.12
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.40
5.2.67
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.67
SAM Revision 3.1
22
November 15, 2007
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Section 5 - Selected Chemical Methods
Analyte
Phosphamidon
Phosphine
Phosphorus trichloride
Pinacolyl methyl phosphonic acid (PMPA)
Propylene oxide
R-33 (VR) [methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-methylpropyl ester]
Sarin
Semivolatile Organic Compounds, NOS
Sodium arsenite
CASRN
13171-21-6
7803-51-2
7719-12-2
616-52-4
75-56-9
159939-87-4
107-44-8
NA
7784-46-5
Method
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
6002 (NIOSH)
6402 (NIOSH)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
1612 (NIOSH)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3571 (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
525.2 (EPA OW)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
200.8 (EPA OW)
3031 (EPA SW-846)
3050B (EPA SW-846)
601 OC (EPA SW-846)
6020A (EPA SW-846)
IO-3.1 (EPAORD)
IO-3.4 (EPA ORD)
IO-3.5 (EPAORD)
Section
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.55
5.2.60
5.2.18
5.2.20
5.2.22
5.2.39
5.2.46
5.2.23
5.2.24
5.2.25
5.2.34
5.2.48
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.21
5.2.35
5.2.46
5.2.10
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.2
5.2.15
5.2.16
5.2.26
5.2.27
5.2.42
5.2.43
5.2.44
SAM Revision 3.1
23
November 15, 2007
-------
Section 5 - Selected Chemical Methods
Analyte
Sodium azide
Soman (GD)
Strychnine
Sulfur dioxide
Sulfur trioxide
Tabun (GA)
Tear gas (CS) [chlorobenzylidene malonitrile]
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Thallium sulfate
CASRN
26628-22-8
96-64-0
57-24-9
7446-09-5
7446-11-g
77-81-6
2698-41-1
107-49-3
80-12-6
10031-59-1
Method
300.1, Rev 1.0 (EPA OW)
3580A (EPA SW-846)
ID-211 (OSHA)
J. of Forensic Science, 1998.
43(1): 200-202
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
6004 (NIOSH)
Method 8 (EPA)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
200.8 (EPA OW)
3031 (EPA SW-846)
3050B (EPA SW-846)
601 OC (EPA SW-846)
6020A (EPA SW-846)
IO-3.1 (EPAORD)
IO-3.4 (EPA ORD)
IO-3.5 (EPAORD)
Section
5.2.5
5.2.22
5.2.68
5.2.81
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.20
5.2.22
5.2.35
5.2.56
5.2.1
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.2
5.2.15
5.2.16
5.2.26
5.2.27
5.2.42
5.2.43
5.2.44
SAM Revision 3.1
24
November 15, 2007
-------
Section 5 - Selected Chemical Methods
Analyte
Thiodiglycol (TDG)
1,4-Thioxane
Titanium tetrachloride
Triacetone triperoxide (TATP)
Triethanolamine (TEA)
Trimethyl phosphite
1,3,5-Trinitrobenzene(1,3,5-TNB)
2,4,6-Trinitrotoluene(2,4,6-TNT)
Vanadium pentoxide
VE [phosphonothioic acid, ethyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VG [phosphonothioic acid, S-(2-
(diethylamino)ethyl) O,O-diethyl ester]
VM [phosphonothioic acid, methyl-,S-(2-
(diethylamino)ethyl) O-ethyl ester]
VX [O-ethyl-S-(2-
diisopropylaminoethyl)methyl-
phosphonothiolate]
CASRN
111-48-8
15980-15-1
7550-45-0
17088-37-8
102-71-6
121-45-9
99-35-4
118-96-7
1314-62-1
21738-25-0
78-53-5
21770-86-5
50782-69-9
Method
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
3050B (EPA SW-846)
601 OC (EPA SW-846)
6020A (EPA SW-846)
3535A (EPA SW-846)
8330B (EPA SW-846)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8321 B (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3535A (EPA SW-846)
8330B (EPA SW-846)
200.8 (EPA OW)
3031 (EPA SW-846)
3050B (EPA SW-846)
601 OC (EPA SW-846)
6020A (EPA SW-846)
IO-3.1 (EPAORD)
IO-3.4 (EPA ORD)
IO-3.5 (EPAORD)
3520C (EPA SW-846)
3535A (EPA SW-846)
3541 (EPA SW-846)
3545A (EPA SW-846)
3580A (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
3571 (EPA SW-846)
8270D (EPA SW-846)
TO-IOA(EPAORD)
Section
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.20
5.2.22
5.2.35
5.2.16
5.2.26
5.2.27
5.2.18
5.2.40
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.39
5.2.46
5.2.18
5.2.20
5.2.22
5.2.35
5.2.46
5.2.18
5.2.40
5.2.2
5.2.15
5.2.16
5.2.26
5.2.27
5.2.42
5.2.43
5.2.44
5.2.17
5.2.18
5.2.19
5.2.20
5.2.22
5.2.35
5.2.46
5.2.21
5.2.35
5.2.46
SAM Revision 3.1
25
November 15, 2007
-------
Section 5 - Selected Chemical Methods
Analyte
White phosphorus
CASRN
12185-10-3
Method
7580 (EPA SW-846)
7905(NIOSH)
Section
5.2.32
5.2.62
The following analytes should be prepared and/or analyzed by the following methods only if problems (e.g.,
insufficient recovery, interferences) occur when using the sample preparation/determinative techniques identified for
these analytes in Appendix A.
Allyl alcohol
3-Chloro-1 ,2-propanediol
Chlorosarin
Chlorosoman
Crimidine
Diisopropyl methylphosphonate (DIMP)
Dimethylphosphoramidic acid
1,4-Dithiane
EA2192 [Diisopropylaminoethyl methyl-
thiophosphonate]
Ethyl methylphosphonic acid (EMPA)
Hydrogen fluoride
Isopropyl methylphosphonic acid (IMPA)
Mercury, Total
Methamidophos
Methoxymercuric acetate
1-Methylethyl ester ethylphosphonofluoridic
acid (GE)
Methylphosphonic acid (MPA)
Perfluoroisobutylene (PFIB)
Pinacolyl methyl phosphonic acid (PMPA)
Sarin
Soman (GD)
1,4-Thioxane
107-18-6
96-24-2
1445-76-7
7040-57-5
535-89-7
1445-75-6
33876-51-6
505-29-3
73207-98-4
1832-53-7
7664-39-3
1832-54-8
7439-97-6
10265-92-6
151-38-2
1189-87-3
993-13-5
382-21-8
616-52-4
107-44-8
96-64-0
15980-15-1
TO-IOA(EPAORD)
TO-15(EPAORD)
TO-15(EPAORD)
8321 B (EPA SW-846)
TO-15(EPAORD)
8270D (EPA SW-846)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
8270D (EPA SW-846)
7906 (NIOSH)
8270D (EPA SW-846)
7470A (EPA SW-846)
7471 B (EPA SW-846)
5600 (NIOSH)
7470A (EPA SW-846)
7471 B (EPA SW-846)
TO-15(EPAORD)
8270D (EPA SW-846)
J. Chrom. A, Vol 1098,2005,
pp. 156-165
8270D (EPA SW-846)
TO-15(EPAORD)
3585 (EPA SW-846)
5030C (EPA SW-846)
5035A (EPA SW-846)
8260C (EPA SW-846)
5.2.46
5.2.47
5.2.47
5.2.39
5.2.47
5.2.35
5.2.23
5.2.24
5.2.25
5.2.34
5.2.35
5.2.63
5.2.35
5.2.29
5.2.30
5.2.52
5.2.29
5.2.30
5.2.47
5.2.35
5.2.80
5.2.35
5.2.47
5.2.23
5.2.24
5.2.25
5.2.34
Method summaries are listed in order of method selection hierarchy (see Figure 2-1), starting with EPA
methods followed by methods from other federal agencies, voluntary consensus standard bodies (VCSB),
and literature references. Methods are listed in numerical order under each publisher. Where available, a
direct link to the full text of the method is provided in the method summary. For additional information
on preparation procedures and methods available through consensus standards organizations, please use
the contact information provided in Table 5-2.
SAM Revision 3.1
26
November 15, 2007
-------
Section 5 - Selected Chemical Methods
Table 5-2. Sources of Chemical Methods
Name
National Environmental Methods Index
(NEMI)
U.S. EPA Office of Water (OW)
Methods
U.S. EPA SW-846 Methods
U.S. EPA Office of Research and
Development Methods
U.S. EPA Air Toxics Methods
Occupational Safety and Health
Administration Methods
National Institutes for Occupational
Safety and Health Methods
Standard Methods for the Examination
of Water and Wastewater, 21st Edition,
2005*
Annual Book of ASTM Standards*
European GESTIS database
International Organization for
Standardization Methods*
Official Methods of Analysis of AOAC
International*
Publisher
EPA, USGS
EPA Office of Water
EPA Office of Solid Waste
and Emergency Response
EPA Office of Research and
Development
EPA Office of Air and
Radiation
OSHA
NIOSH
American Public Health
Association (APHA),
American Water Works
Association (AWWA), and
Water Environment
Federation (WEF)
ASTM International
HVBG
ISO
AOAC International
Reference
h|JE//wwjiemLgov
http://vwvw.epa.qov/safewater/methods/
sourcalt.html
http://vwvw.epa.qov/epaoswer/hazwaste
/test/mainjTtm
http://www.epa.gov/nerlcwww/ordmeth.
Mm
Htj)Jjvwm__:_ej)_a._Q_a^
http://www.osha.qov
http://www.cdc.qov/niosh/nmam/
http://www.standardmethods.orq
http://www.astm.orq
http://www.hvbq. de/e/bia/qestis/analytic
al methods/index. html
http://www.iso.orq
http://www.aoac.orq
' Subscription and/or purchase required.
5.1.2 General Quality Control (QC) Guidance for Chemical Methods
Having analytical data of appropriate quality requires that laboratories: (1) conduct the necessary QC to
ensure that measurement systems are in control and operating correctly, (2) properly document results of
the analyses, and (3) properly document measurement system evaluation of the analysis-specific QC,
including corrective actions. In emergency situations, however, speed and efficiency are also important.
Laboratories must be prepared with calibrated instruments, the proper standards, standard analytical
procedures, and qualified and trained technicians. Laboratories also must be capable of providing rapid
turnaround of sample analyses and data reporting.
The level or amount of QC needed during sample analysis and reporting depends on the intended purpose
of the data that are generated (i.e., the decision(s) to be made). The specific needs for data generation
should be identified. QC requirements and data quality objectives should be derived based on those
needs, and should be applied consistently across laboratories when multiple laboratories are used. For
almost all of the chemical warfare agents, most laboratories will not have access to analytical standards
for calibration and quality control. Use of these agents is strictly controlled by the DoD and access is
SAM Revision 3.1
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Section 5 - Selected Chemical Methods
limited. For information regarding purchase and distribution of ultradilute agents, please contact Terry
Smith, EPA's Office of Emergency Management, at (202) 564-2908.
A minimum set of analytical QC procedures should be planned and conducted for all chemical testing.
Some method-specific QC requirements are described in many of the individual methods that are cited in
this document and will be referenced in any Standardized Analytical Protocols (SAPs) developed to
address specific analytes and sample types of concern. Individual methods, sampling and analysis
protocols, or contractual statements of work also should be consulted to determine any additional QC that
may be needed. Analytical QC requirements generally consist of analysis of laboratory control samples
to document whether the analytical system is in control, matrix spikes (MS) to identify and quantify
measurement system accuracy for the media of concern and at the levels of concern, various blanks as a
measure of freedom from contamination, and matrix spike duplicates (MSB) or sample replicates to
assess data precision.
In general, for measurement of chemical analytes, appropriate QC includes an initial demonstration of
measurement system capability as well as ongoing analysis of standards and other samples to ensure the
continued reliability of the analytical results. Examples of appropriate quality control include:
Demonstration that measurement system is operating properly:
>• Initial calibration; and
>• Method blanks.
Demonstration of analytical method suitability for intended use:
>• Detection and quantitation limits;
>• Precision and recovery (verify measurement system has adequate accuracy); and
>• Analyte/matrix/level of concern-specific QC samples (verify that measurement system has
adequate sensitivity at levels of concern).
Demonstration of continued analytical method reliability:
>• Matrix spike/matrix spike duplicates (recovery and precision);
>• QC samples (system accuracy and sensitivity at levels of concern);
>• Surrogate spikes (where appropriate);
>• Continuing calibration verification; and
>• Method blanks.
QC tests should be run as frequently as necessary to ensure the reliability of analytical results. As with
the identification of needed QC samples, frequency should be established based on an evaluation of data
quality objectives. The type and frequency of QC can be focused over time.
Ensuring data quality also requires that laboratory results are properly evaluated and the results of the data
quality evaluation are transmitted to decision makers. This evaluation is as important as the data in
ensuring informed and effective decisions. While some degree of data evaluation is necessary in order to
be able to confirm data quality, 100% verification and/or validation is neither necessary nor conducive to
efficient decision making in emergency situations. The level of such reviews should be determined based
on the specific situation being assessed and on the corresponding data quality objectives. In every case,
the levels of QC and data review necessary to support decision making should be determined as much in
advance of data collection as possible.
Please note: The appropriate point of contact identified in Section 4 should be consulted regarding
appropriate quality assurance and quality control (QA/QC) procedures prior to sample analysis. These
contacts will consult with the EPA Office of Solid Waste and Emergency Response (OSWER)
coordinator responsible for laboratory activities during the specific event to ensure QA/QC procedures are
performed consistently across laboratories. OSWER is planning to develop QA/QC guidance for
laboratory support. EPA program offices will be responsible for ensuring that the QA/QC practices are
implemented.
SAM Revision 3.1 28 November 15, 2007
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Section 5 - Selected Chemical Methods
5.1.3 Safety and Waste Management
It is imperative that safety precautions are used during collection, processing, and analysis of
environmental samples. Laboratories should have a documented health and safety plan for handling
samples that may contain the target chemical, biological, or radiological contaminants, and laboratory
staff should be trained in and implement the safety procedures included in the plan. In addition, many of
the methods summarized or cited in Section 5.2 contain some specific requirements, guidance, or
information regarding safety precautions that should be followed when handling or processing
environmental samples and reagents. These methods also provide information regarding waste
management. Other resources that can be consulted for additional information include the following:
• Occupational Health and Safety Administration's (OSHA) standard for Occupational Exposure to
Hazardous Chemicals in Laboratories (29 CFR 1910.1450)
• OSHA regulations for hazardous waste operations and emergency response (29 CFR 1910)
• Environmental Protection Agency's standards regulating hazardous waste (40 CFR parts 260 - 270)
• U.S. Department of Transportation (DOT) regulations for transporting hazardous materials (49 CFR
Part 172)
• U.S. Department of Health and Human Services, Centers for Disease Control and Prevention's
requirements for possession, use, and transfer of select agents and toxins (42 CFR Part 1003)
5.2 Method Summaries
Summaries for the analytical methods listed in Appendix A are provided in Sections 5.2.1 through 5.2.81.
These sections contain summary information only, extracted from the selected methods. Each method
summary contains a table identifying the contaminants in Appendix A to which the method applies, a
brief description of the analytical method, and a link to or source for obtaining a full version of the
method. The full version of the method should be consulted prior to sample analysis.
5.2.1 EPA Method 8: Determination of Sulfuric Acid and Sulfur Dioxide Emissions from
Stationary Sources
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Sulfur Trioxide
CASRN
7446-11-9
A gas sample is extracted isokinetically. Sulfuric acid (H2SO4) and sulfur dioxide (SO2) are separated,
and both fractions are measured separately by the barium-thorin titration method. Sulfur trioxide (SO3) is
measured by the analysis of H2SO4. Possible interfering agents include fluorides, free ammonia, and
dimethyl aniline. Collaborative tests have shown that the minimum detectable limits of the method are
0.06 mg/m3 for H2SO4 mist/ SO3.
Source: EPA Method 8: Determination Of Sulfuric Acid and Sulfur Dioxide Emissions From Stationary
Sources. EPA Emission Measurement Center (EMC) of the Office of Air Quality Planning and Standards
(OAQPS). http://www.epa.gov/ttnemc01/methods/method8.html
SAM Revision 3.1 29 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.2 EPA Method 200.8: Determination of Trace Elements in Waters and Wastes by
Inductively Coupled Plasma-Mass Spectrometry
This method should be used for preparation and analysis of aqueous liquid and drinking water samples
for the contaminants identified below and listed in Appendix A.
Analyte(s)
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
Arsine
2-Chlorovinylarsonous acid (CVAA)
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]1
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite Oxide
Sodium arsenite
Thallium sulfate
Vanadium pentoxide
CASRN
7803-55-6
740-38-2
1327-53-3
7784-42-1
85090-33-1
541-25-3
40334-69-8
40334-70-1
1306-02-1
7784-46-5
10031-59-1
1314-62-1
1 Laboratory testing is currently underway for speciation of lewisite 1 using GC-MS techniques.
This method will determine metal-containing compounds only as the total metal (e.g., total arsenic). An
aliquot of a we 11-mixed, homogeneous sample is accurately weighed or measured for sample processing.
For total recoverable analysis of a sample containing undissolved material, analytes are first solubilized
by gentle refluxing with nitric and hydrochloric acids. After cooling, the sample is made up to volume,
mixed, and centrifuged or allowed to settle overnight prior to analysis. For determination of dissolved
analytes in a filtered aqueous sample aliquot, or for the "direct analysis" total recoverable determination
of analytes in drinking water where sample turbidity is < 1 NTU, the sample is made ready for analysis by
the addition of nitric acid, and then diluted to a predetermined volume and mixed before analysis. The
prepared sample is analyzed using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). Specific
analytes targeted by Method 200.8 are listed in Section 1.1 of the method. The recommended calibration
range is 10 to 200 (ig/L. Method detection limits for arsenic in aqueous samples have been found to be
1.4 (ig/L in scanning mode, and 0.4 (ig/L in selected ion monitoring mode.
Source: EPA Method 200.8: Determination of Trace Elements in Waters and Wastes by Inductively
Coupled Plasma-Mass Spectrometry, Revision 5.4, 1994.
http://infotrek.er.usgs.gov/pls/apex/f?D=l 19:38:1701708976536019::::P38 METHOD ID:4665
SAM Revision 3.1
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Section 5 - Selected Chemical Methods
5.2.3 EPA Method 245.2: Mercury (Automated Cold Vapor Technique)
This method should be used for preparation and analysis of drinking water samples for the contaminants
identified below and listed in Appendix A.
Analyte(s)
Mercury, Total
Methoxyethylmercuric acetate
CASRN
7439-97-6
151-38-2
This method will determine methoxyethylmercuric acetate as total mercury. If dissolved mercury is
targeted, the sample is filtered prior to acidification. To detect total mercury (inorganic and organic
mercury), the sample is treated with potassium permanganate and potassium persulfate to oxidize organic
mercury compounds prior to analysis. Inorganic mercury is reduced to the elemental state (using tin
sulfate or tin chloride) and aerated from solution. The mercury vapor passes through a cell positioned in
the light path of a cold vapor atomic absorption (CVAA) spectrophotometer. The concentration of
mercury is measured using the CVAA spectrophotometer. Applicable concentration range is 0.2 to 20.0
jig/L.
Source: EPA Method 245.2: Mercury (Automated Cold Vapor Technique), 1974.
http://infotrek.er.usgs.gov/pls/apex/ff p= 119:38:1701708976536019:::: P38 M ETHOP I P:4822
5.2.4 EPA Method 252.2: Osmium (Atomic Absorption, Furnace Technique)
This method should be used for the preparation and analysis of aqueous liquid and drinking water
samples for the contaminant identified below and listed in Appendix A.
Analyte(s)
Osmium tetroxide
CASRN
20816-12-0
This method will determine osmium tetroxide as osmium. Method 252.2 is not a stand alone method in
that sections of the method reference "Methods of Chemical Analysis of Water and Waste", EPA/600/4-
79/020, March 1983 (MCAWW). Samples are prepared according to the "direct aspiration method" (See
Section 9.1 of the Atomic Absorption Methods section of MCAWW) except that the addition of sulfuric
acid is omitted in the final adjustment. If only dissolved osmium is determined, the sample is filtered
before acidification with nitric acid. For total osmium, the sample is digested with nitric and hydrochloric
acids and made up to volume. Samples are analyzed according to the "furnace procedure" (see Section
9.3 of the Atomic Absorption Methods section of MCAWW), using a graphite furnace atomic absorption
spectrometer. A representative aliquot of sample is placed in the graphite tube in the furnace, evaporated
to dryness, chaffed, and atomized. Radiation from an excited element is passed through the vapor
containing ground state atoms of the element. The intensity of the transmitted radiation decreases in
proportion to the amount of the ground state element in the vapor. A monochromator isolates the
characteristic radiation from the hollow cathode lamp and a photosensitive device measures the attenuated
transmitted radiation. The optimal applicable concentration range is 50 to 500 (ig/L. Detection limits for
osmium using this method have been found to be 20 (ig/L.
Source: EPA Method 252.2: Osmium (AA, Furnace Technique), 1978.
ID:1199
SAM Revision 3.1 31 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.5 EPA Method 300.1, Revision 1.0: Determination of Inorganic Anions in Drinking
Water by Ion Chromatography
This method should be used for the preparation and analysis of aqueous liquid and drinking water
samples for the contaminants identified below and listed in Appendix A. It also should be used for
analysis of solid, non-aqueous liquid/organic solid, and air samples for fluoroacetic acid, fluoroacetate
salts, and methyl fluoroacetate when appropriate sample preparation techniques have been applied (refer
to Appendix A).
Analyte(s)
Fluoride
Fluoroacetic acid and fluoroacetate salts
Methyl fluoroacetate
CASRN
16984-48-8
NA
453-18-9
This method was developed for analysis of aqueous samples, and can be adapted for analysis of prepared
non-aqueous liquid/organic solid, solid, and air samples when appropriate sample preparation techniques
have been applied (see Appendix A). A small volume of an aqueous liquid sample (10 (iL or 50 (iL) is
introduced into an ion chromatograph. The volume selected depends on the concentration of
fluoroacetate ion in the sample. The anions of interest are separated and measured, using a system
comprising a guard column, analytical column, suppressor device, and conductivity detector. The
separator columns and guard columns, as well as eluent conditions, are identical. To achieve comparable
detection limits, an ion chromatographic system must use suppressed conductivity detection, be properly
maintained, and be capable of yielding a baseline with no more than 5 nS noise/drift per minute of
monitored response over the background conductivity. The method detection limit varies depending upon
the nature of the sample and the specific instrumentation employed. The estimated calibration range is
approximately two orders of magnitude. The detection limit for fluoride in reagent water is 0.009 mg/L.
Source: EPA Method 300.1: Determination of Inorganic Anions in Drinking Water by Ion
Chromatography, Revision 1.0, 1997.
IP:4674
5.2.6 EPA Method 335.4: Determination of Total Cyanide by Semi-Auto mated
Colorimetry
This method should be used for preparation and analysis of drinking water samples for the contaminants
identified below and listed in Appendix A.
Analyte(s)
Cyanide, Total
Hydrogen cyanide
CASRN
57-12-5
74-90-8
Cyanide is released from cyanide complexes as hydrocyanic acid by manual reflux-distillation, and
absorbed in a scrubber containing sodium hydroxide solution. The cyanide ion in the absorbing solution
is converted to cyanogen chloride by reaction with chloramine-T, which subsequently reacts with pyridine
and barbituric acid to give a red-colored complex. Some interferences, such as aldehydes, nitrate-nitrite,
oxidizing agents, thiocyanate, thiosulfate, and sulfide, are eliminated or reduced by distillation. The
applicable range is 5 to 500 (ig/L.
Source: EPA Method 335.4: Determination of Total Cyanide by Semi-Automated Colorimetry, Revision
1.0, 1993.
http://infotrek.er.usgs.gov/pls/apex/f?p=l 19:38:1701708976536019::::P38_METHOD_ID:5759
SAM Revision 3.1 32 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.7 EPA Method 350.1: Nitrogen, Ammonia (Colorimetric, Automated Phenate)
This method should be used for preparation and analysis of drinking water samples for the contaminant
identified below and listed in Appendix A.
Analyte(s)
Ammonia
CASRN
7664-41-7
This method identifies and determines the concentration of ammonia in drinking water samples by
spectrophotometry. Samples are buffered at a pH of 9.5 with borate buffer to decrease hydrolysis of
cyanates and organic nitrogen compounds, and are distilled into a solution of boric acid. Alkaline phenol
and hypochlorite react with ammonia to form indophenol blue that is proportional to the ammonia
concentration. The blue color formed is intensified with sodium nitroprusside and measured
spectrophotometrically. The working range for ammonia is 0.01 to 2.0 mg/L.
Source: EPA Method 350.1: Nitrogen, Ammonia (Colorimetric, Automated Phenate), Revision 2.0, 1993.
http://infotrok.cr.usgs.gov/pls/apcx/f?p=119:38:1701708976536019::::P38
5.2.8 EPA Method 507: Determination of Nitrogen- and Phosphorus-Containing
Pesticides in Water by Gas Chromatography with a Nitrogen-Phosphorus Detector
This method should be used for preparation and analysis of drinking water samples for the contaminant
identified below and listed in Appendix A.
Analyte(s)
Organophosphate Pesticides, NOS1
CASRN
NA
NOS = Not otherwise specified
This is a gas chromatographic (GC) method applicable to the determination of certain nitrogen and
phosphorus containing pesticides in ground water and finished drinking water. A 1-L sample is extracted
with methylene chloride by shaking in a separatory funnel or mechanical tumbling in a bottle. The
methylene chloride extract is isolated, dried, and concentrated to a volume of 5 mL during a solvent
exchange to methyl tert-butyl ether (MTBE). The concentrations of pesticides in the extract are measured
using a capillary column GC system equipped with a nitrogen-phosphorus detector (NPD). Specific
analytes targeted by Method 507 are listed in Section 1.1 of the method. Resulting estimated detection
limits and method detection limits (MDLs) differ depending on the specific pesticide.
Please note: The presence of any organophosphate pesticide must be confirmed by either a secondary GC
column or by a mass spectrometer.
Source: EPA Method 507: Determination of Nitrogen- and Phosphorus-Containing Pesticides in Water by
Gas Chromatography with a Nitrogen-Phosphorus Detector, Revision 2.1, 1995.
http://iiifotrok.cr.usgs.gov/pls/apcx/f?p=119:38:1701708976536019::::P38
SAM Revision 3.1 33 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.9 EPA Method 524.2: Measurement of Purgeable Organic Compounds in Water by
Capillary Column Gas Chromatography / Mass Spectrometry
This method should be used for preparation and analysis of drinking water samples for the contaminants
identified below and listed in Appendix A.
Analyte(s)
Carbon disulfide
1,2-Dichloroethane
CASRN
75-15-0
107-06-2
Volatile organic compounds and surrogates with low water solubility are extracted (purged) from the
sample matrix by bubbling an inert gas through the aqueous sample. Purged sample components are
trapped in a tube containing suitable sorbent materials. When purging is complete, the sorbent tube is
heated and backflushed with helium to desorb the trapped sample components into a capillary gas
chromatography (GC) column interfaced to a mass spectrometer (MS). The column is temperature
programmed to facilitate the separation of the method analytes, which are then detected with the MS.
Specific analytes targeted by Method 524.2 are listed in Section 1.1 of the method. Detection levels for
carbon disulfide and 1,2-dichloroethane in reagent water have been found to be 0.093 (ig/L and 0.02
(ig/L, respectively.
Source: EPA Method 524.2: Measurement of Purgeable Organic Compounds in Water by Capillary
Column Gas Chromatography/Mass Spectrometry, Revision 4.0, 1992.
http://infotrok.cr.usgs.gov/pls/apcx/f?p=119:38:1701708976536019::::P38
5.2.10 EPA Method 525.2: Determination of Organic Compounds in Drinking Water by
Liquid-Solid Extraction and Capillary Column Gas Chromatography / Mass
Spectrometry
This method should be used for preparation and analysis of drinking water samples for the contaminants
identified below and listed in Appendix A.
Analyte(s)
Dichlorvos
Fenamiphos
Mevinphos
Semivolatile Organic Compounds, NOS1
CASRN
62-73-7
22224-92-6
7786-34-7
NA
NOS = Not otherwise specified
Organic compounds, internal standards, and surrogates are extracted from a water sample by passing 1 L
of sample through a cartridge or disk containing a solid matrix with chemically bonded Ci8 organic phase
(liquid-solid extraction, LSE or solid-phase extraction, SPE). The organic compounds are eluted from the
LSE (SPE) cartridge or disk with small quantities of ethyl acetate followed by methylene chloride. The
resulting extract is concentrated further by evaporation of some of the solvent. Sample components are
separated, identified, and measured by injecting an aliquot of the concentrated extract into a high
resolution fused silica capillary column of a gas chromatography/mass Spectrometry (GC-MS) system.
Specific analytes targeted by Method 525.2 are listed in Section 1.1 of the method. The applicable
concentration range for most analytes is 0.1 to 10 (ig/L.
Source: EPA Method 525.2: Determination of Organic Compounds in Drinking Water by Liquid-Solid
Extraction and Capillary Column Gas Chromatography/Mass Spectrometry, Revision 2.0, 1995.
http://infotrek.er.usgs.gov/pls/apex/f?p= 119:38:1701708976536019:::: P38 M 10:4804
SAM Revision 3.1 34 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.11 EPA Method 531.2: Measurement of N-Methylcarbamoyloximes and N-
Methylcarbamates in Water by Direct Aqueous Injection HPLC with Postcolumn
Derivatization
This method should be used for preparation and analysis of drinking water samples for the contaminants
identified below and listed in Appendix A.
Analyte(s)
Aldicarb (Temik)
Carbofuran (Furadan)
Methomyl
Oxamyl
CASRN
116-06-3
1563-66-2
16752-77-5
23135-22-0
An aliquot of sample is measured in a volumetric flask. Samples are preserved, spiked with appropriate
surrogates and then filtered. Analytes are chromatographically separated by injecting a sample aliquot
(up to 1000 (iL) into a high performance liquid chromatographic (HPLC) system equipped with a reverse
phase (Ci8) column. After elution from the column, the analytes are hydrolyzed in a post column reaction
to form methylamine, which is in turn reacted to form a fluorescent isoindole that is detected by a
fluorescence detector. Analytes also are quantitated using the external standard technique.
Source: EPA Method 531.2: Measurement of N-Methylcarbamoyloximes and N-Methylcarbamates in
Water by Direct Aqueous Injection HPLC with Postcolumn Derivitization, Revision 1.0, 2001.
http://iiifotrck.cr.usgs.gov/pls/apcx/f?p=119:38:1701708976536019::::P38 ID:7392
5.2.12 EPA Method 549.2: Determination of Diquat and Paraquat in Drinking Water by
Liquid-Solid Extraction and High-Performance Liquid Chromatography with
Ultraviolet Detection
This method should be used for preparation and analysis of aqueous liquid and drinking water samples
for the contaminant identified below and listed in Appendix A.
Analyte(s)
Paraquat
CASRN
4685-14-7
A 250-mL sample is extracted using a C8 liquid/solid extraction (LSE) cartridge or a C8 disk that has been
specially prepared for the reversed-phase, ion-pair mode. The LSE disk or cartridge is eluted with acidic
aqueous solvent to yield the eluate/extract. An ion-pair reagent is added to the eluate/extract. The
concentrations of paraquat in the eluate/extract are measured using a high performance liquid
chromatography (HPLC) system equipped with a UV absorbance detector. A photodiode array detector is
used to provide simultaneous detection and confirmation of the method analytes. The analytical range
depends on the sample matrix and the instrumentation used.
Source: EPA Method 549.2: Determination of Diquat and Paraquat in Drinking Water by Liquid-Solid
Extraction and High Performance Liquid Chromatography with Ultraviolet Detection, Revision 1.0, 1997.
http://www.cpa.gOv/ncrlcwww/m 549 2.pdf
SAM Revision 3.1 35 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.13 EPA Method 551.1: Determination of Chlorination Disinfection Byproducts,
Chlorinated Solvents, and Halogenated Pesticides/Herbicides in Drinking Water by
Liquid-Liquid Extraction and Gas Chromatography with Electron-Capture
Detection
This method should be used for preparation and analysis of drinking water samples for the contaminant
identified below and listed in Appendix A.
Analyte(s)
Chloropicrin
CASRN
79-06-2
This is a gas chromatographic/electron capture detection (GC-ECD) method applicable to the
determination of halogenated analytes in finished drinking water, drinking water during intermediate
stages of treatment, and raw source water. A 50-mL sample aliquot is extracted with 3 mL of methyl tert-
butyl ether (MTBE) or 5 mL of pentane. Two uL of the extract is then injected into a GC equipped with a
fused silica capillary column and linearized electron capture detector for separation and analysis. This
liquid/liquid extraction technique efficiently extracts a wide boiling range of non-polar and polar organic
components of the sample. Thus, confirmation is quite important, particularly at lower analyte
concentrations. A confirmatory column is suggested for this purpose. The Estimated Detection Limit
(EDL) using MTBE and ammonium chloride-preserved reagent water on a DB-1 column has been found
tobe0.014(ig/L.
Source: EPA Method 551.1: Determination of Chlorination Disinfection Byproducts, Chlorinated
Solvents, and Halogenated Pesticides/Herbicides in Drinking Water by Liquid-Liquid Extraction and Gas
Chromatography with Electron-Capture Detection.
http://iiifotrck.cr.usgs.gov/pls/apcx/f?p=119:38:1701708976536019::::P38
5.2.14 EPA Method 614: The Determination of Organophosphorus Pesticides in
Municipal and Industrial Wastewater
This method should be used for preparation and analysis of aqueous liquid samples for the contaminant
identified below and listed in Appendix A.
Analyte(s)
Organophosphate Pesticides, NOS1
CASRN
NA
NOS = Not otherwise specified
This is a gas chromatographic (GC) method applicable to the determination of organophosphate
pesticides in industrial and municipal discharges using a GC with a phosphorus-specific flame
photometric detector (FPD) or thermionic bead detector in the nitrogen mode. A measured volume of
sample, approximately 1 L, is extracted with 15% methylene chloride in hexane using a separatory funnel.
The extract is dried and concentrated to a volume of 10 mL or less. Gas chromatographic conditions are
described for the separation and measurement of the compounds in the extract by flame photometric or
thermionic bead gas Chromatography. Specific analytes targeted by Method 614 are listed in Section 1.1
of the method.
Please note: The presence of any organophosphate pesticide must be confirmed by either a secondary GC
solumn or with a mass spectrometer.
SAM Revision 3.1 36 November 15, 2007
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Section 5 - Selected Chemical Methods
Source: "Methods for the Determination of Nonconventional Pesticides in Municipal and Industrial
Wastewater - Vol. I," United States Environmental Protection Agency, available from National Technical
Information Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161. Phone: 800-553-6847.
5.2.15 EPA Method 3031 (SW-846): Acid Digestion of Oils for Metals Analysis by Atomic
Absorption or ICP Spectrometry
This method should be used for preparation of non-aqueous liquid/organic solid samples for the
contaminants identified below and listed in Appendix A. Refer to Appendix A for the appropriate
determinative method for these analytes.
Analyte(s)
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
2-Chlorovinylarsonous acid (CVAA)
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite Oxide
Sodium arsenite
Thallium sulfate
Vanadium pentoxide
CASRN
7803-55-6
740-38-2
1327-53-3
85090-33-1
541-25-3
40334-69-8
40334-70-1
1306-02-1
7784-46-5
10031-59-1
1314-62-1
This method is used to prepare samples for the determination of arsenic trioxide, lewisite, lewisite
degradation products, and sodium arsenite as total arsenic; thallium sulfate as total thallium; and
ammonium metavanadate and vanadium pentoxide as total vanadium. A 0.5-g sample of oil, oil sludge,
tar, wax, paint, or paint sludge is mixed with potassium permanganate and sulfuric acid. The mixture is
then treated with nitric and hydrochloric acids, filtered, and diluted to volume. Excess manganese may be
removed with ammonium hydroxide. Digestates are analyzed by Method 6020A or 6010C (SW-846).
Source: EPA Method 3031 (SW-846): Acid Digestion of Oils for Metals Analysis by Atomic Absorption
or ICP Spectrometry, Revision 0, 1996. http://www.cpa.gov/cpaoswcr/hazwastc/tcst/pdfs/3031.pdf
5.2.16 EPA Method 3050B (SW-846): Acid Digestion of Sediments, Sludges, and Soils
This method should be used for preparation of solid samples for the contaminants identified below and
listed in Appendix A. Refer to Appendix A for the appropriate determinative method for these analytes.
Analyte(s)
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
Arsine
CASRN
7803-55-6
740-38-2
1327-53-3
7784-42-1
SAM Revision 3.1
37
November 15, 2007
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Section 5 - Selected Chemical Methods
Analyte(s)
2-Chlorovinylarsonous acid (CVAA)
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite Oxide
Osmium tetroxide
Sodium arsenite
Thallium sulfate
Titanium tetrachloride
Vanadium pentoxide
CASRN
85090-33-1
541-25-3
40334-69-8
40334-70-1
1306-02-1
20816-12-0
7784-46-5
10031-59-1
7550-45-0
1314-62-1
This method is used to prepare samples for the determination of arsenic trioxide, arsine, lewisite, lewisite
degradation products, and sodium arsenite as total arsenic; thallium sulfate as total thallium; titanium
tetrachloride as titanium; osmium tetroxide as osmium; and ammonium metavanadate and vanadium
pentoxide as total vanadium. A 1-g to 2-g sample is digested with nitric acid and hydrogen peroxide.
Sample volumes are reduced, then brought up to a final volume of 100 mL. Samples are analyzed for
total arsenic, total thallium, total titanium, or total vanadium by Method 6010C or 6020A (SW-846); use
Method 60 IOC (SW-846) for total osmium.
Source: EPA Method 3050B (SW-846): Acid Digestion of Sediments, Sludges, and Soils, Revision 2,
1996. httPi/AvwwxjpcLgOT/ej^^
5.2.17 EPA Method 3520C (SW-846): Continuous Liquid-Liquid Extraction
This method should be used for preparation of aqueous liquid and drinking water samples for the
contaminants identified below and listed in Appendix A. Note: For fenamiphos and semivolatile organic
compounds only, EPA Method 525.2 (rather than Method 3520C) should be used for preparation of
drinking water samples. Refer to Appendix A for the appropriate determinative method for these
analytes.
Analyte(s)
Brodifacoum
Bromadiolone
Chlorfenvinphos
3-Chloro-1 ,2-propanediol
Chlorosarin
Chlorosoman
Chlorpyrifos
Cyclohexyl sarin (GF)
Diesel Range Organics
CASRN
56073-10-0
28772-56-7
470-90-6
96-24-2
1445-76-7
7040-57-5
2921-88-2
329-99-7
NA
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Section 5 - Selected Chemical Methods
Analyte(s)
Diphacinone
N-Ethyldiethanolamine (EDEA)
Fenamiphos
Fentanyl
Methyl hydrazine
N-Methyldiethanolamine (MDEA)
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Mustard, nitrogen (HN-1) [bis(2-chloroethyl)ethylamine]
Mustard, nitrogen (HN-2) [2,2'-dichloro-N-
methyldiethylamine N,N-bis(2-chloroethyl)methylamine]
Mustard, nitrogen (HN-3) [tris(2-chloroethyl)amine]
Parathion
Phenol
Phosphamidon
R-33 (VR) [methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-methylpropyl ester]
Semivolatile Organic Compounds, NOS1
Tear gas (CS) [chlorobenzylidene malonitrile]
Tetramethylenedisulfotetramine
Triethanolamine (TEA)
VE [phosphonothioic acid, ethyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VG [phosphonothioic acid, S-(2-(diethylamino)ethyl)
O,O-diethyl ester]
VM [phosphonothioic acid, methyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
CASRN
82-66-6
139-87-7
22224-92-6
437-38-7
60-34-4
105-59-9
1189-87-3
538-07-8
51-75-2
555-77-1
56-38-2
108-95-2
13171-21-6
159939-87-4
NA
2698-41-1
80-12-6
102-71-6
21738-25-0
78-53-5
21770-86-5
1 NOS = Not otherwise specified
This method is applicable to the isolation and concentration of water-insoluble and slightly soluble
organics in preparation for a variety of chromatographic procedures. A measured volume of sample,
usually 1 L, is placed into a continuous liquid-liquid extractor, adjusted, if necessary, to a specific pH and
extracted with organic solvent for 18 to 24 hours. The extract is filtered through sodium sulfate to
remove residual moisture, concentrated, and exchanged as necessary into a solvent compatible with the
cleanup or determinative procedure used for analysis.
Source: EPA Method 3520C (SW-846): Continuous Liquid-Liquid Extraction, Revision 3, 1996.
http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3520c.pdf
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Section 5 - Selected Chemical Methods
5.2.18 EPA Method 3535A (SW-846): Solid-Phase Extraction
This method should be used for preparation of aqueous liquid and/or drinking water samples for the
contaminants identified below and listed in Appendix A. Note: For dichlorvos, fenamiphos, mevinphos,
and semivolatile organic compounds only, EPA Method 525.2 (rather than Method 3535A) should be
used for preparation of drinking water samples. For poly chlorinated biphenyls only, EPA Method 508
(rather than Method 3535 A) should be used for preparation of drinking water samples. Refer to
Appendix A for the appropriate determinative method for these analytes.
Analyte(s)
2-Amino-4,6-dinitrotoluene (2-Am-DNT)
4-Amino-2,6-dinitrotoluene (4-Am-DNT)
4-Aminopyridine
Brodifacoum
Bromadiolone
Chlorfenvinphos
3-Chloro-1 ,2-propanediol
Chlorosarin
Chlorosoman
Chlorpyrifos
Crimidine
Cyclohexyl sarin (GF)
Dichlorvos
Dicrotophos
Diesel Range Organics
Diisopropyl methylphosphonate (DIMP)
Dimethylphosphite
Dimethylphosphoramidic acid
3,5-Dinitroaniline (3,5-DNA)
1,3-Dinitrobenzene (1,3-DNB)
2,4-Dinitrotoluene (2,4-DNT)
2,6-Dinitrotoluene (2,6-DNT)
Diphacinone
1,4-Dithiane
EA2192 [Diisopropylaminoethyl
methylthiolophosphonate]
Ethyl methylphosphonic acid (EMPA)
Ethyldichloroarsine (ED)
CASRN
35572-78-2
19406-51-0
504-24-5
56073-10-0
28772-56-7
470-90-6
96-24-2
1445-76-7
7040-57-5
2921-88-2
535-89-7
329-99-7
62-73-7
141-66-2
NA
1445-75-6
868-85-9
33876-51-6
618-87-1
99-65-0
121-14-2
606-20-2
82-66-6
505-29-3
73207-98-4
1832-53-7
598-14-1
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Section 5 - Selected Chemical Methods
Analyte(s)
N-Ethyldiethanolamine (EDEA)
Fenamiphos
Fentanyl
Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)
Hexamethylenetriperoxidediamine (HMTD)
Isopropyl methylphosphonic acid (IMPA)
Methamidophos
Methyl hydrazine
Methyl parathion
Methyl-2,4,6-trinitrophenylnitramine (Tetryl)
N-Methyldiethanolamine (MDEA)
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Methylphosphonic acid (MPA)
Mevinphos
Mustard, nitrogen (HN-1) [bis(2-chloroethyl)ethylamine]
Mustard, nitrogen (HN-2) [2,2'-dichloro-N-
methyldiethylamine N,N-bis(2-chloroethyl)methylamine]
Mustard, nitrogen (HN-3) [tris(2-chloroethyl)amine]
Nicotine sulfate
Nitrobenzene (NB)
Nitroglycerin (NG)
2-Nitrotoluene (2-NT)
3-Nitrotoluene (3-NT)
4-Nitrotoluene (4-NT)
Octahydro-1 ,3,5,7-tetranitro-1 ,3,5,7-tetrazocine (HMX)
Parathion
Pentaerythritol tetranitrate (PETN)
Phencyclidine
Phenol
Phorate
Phosphamidon
Pinacolyl methyl phosphonic acid (PMPA)
CASRN
139-87-7
22224-92-6
437-38-7
121-82-4
283-66-9
1832-54-8
10265-92-6
60-34-4
298-00-0
479-45-8
105-59-9
1189-87-3
993-13-5
7786-34-7
538-07-8
51-75-2
555-77-1
54-11-5
98-95-3
55-63-0
88-72-2
99-08-1
99-99-0
2691-41-0
56-38-2
78-11-5
77-10-1
108-95-2
298-02-2
13171-21-6
616-52-4
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Section 5 - Selected Chemical Methods
Analyte(s)
R-33 (VR) [methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-methylpropyl ester]
Semivolatile Organic Compounds, NOS1
Soman (GD)
Strychnine
Tabun (GA)
Tear gas (CS) [chlorobenzylidene malonitrile]
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine2
Thiodiglycol (TDG)
1,4-Thioxane
Triacetone triperoxide (TATP)
Triethanolamine (TEA)
Trimethyl phosphite
1 ,3,5-Trinitrobenzene (1 ,3,5-TNB)
2,4,6-Trinitrotoluene(2,4,6-TNT)
VE [phosphonothioic acid, ethyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VG [phosphonothioic acid, S-(2-(diethylamino)ethyl)
O,O-diethyl ester]
VM [phosphonothioic acid, methyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
CASRN
159939-87-4
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
111-48-8
15980-15-1
17088-37-8
102-71-6
121-45-9
99-35-4
118-96-7
21738-25-0
78-53-5
21770-86-5
1 NOS = Not otherwise specified
2 This analyte may require SPE extraction using acetone or methyl ethylketone
This method describes a procedure for isolating target organic analytes from aqueous and liquid samples
using solid-phase extraction (SPE) media. Sample preparation procedures vary by analyte group.
Following any necessary pH adjustment, a measured volume of sample is extracted by passing it through
the solid-phase extraction medium (disks or cartridges), which is held in an extraction device designed for
vacuum filtration of the sample. Target analytes are eluted from the solid-phase media using an
appropriate solvent which is collected in a receiving vessel. The resulting solvent extract is dried using
sodium sulfate and concentrated, as needed.
Source: EPA Method 3535A (SW-846): Solid-Phase Extraction (SPE), Revision 1, 1998.
http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3535a.pdf
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Section 5 - Selected Chemical Methods
5.2.19 EPA Method 3541 (SW-846): Automated Soxhlet Extraction
This method should be used for preparation of solid
listed in Appendix A. Refer to Appendix A for the a
Analyte(s)
Brodifacoum
Bromadiolone
Chlorfenvinphos
3-Chloro-1 ,2-propanediol
Chlorosarin
Chlorosoman
Chlorpyrifos
Cyclohexyl sarin (GF)
Diesel Range Organics
Diphacinone
N-Ethyldiethanolamine (EDEA)
Fenamiphos
Fentanyl
Methamidophos
Methyl hydrazine
N-Methyldiethanolamine (MDEA)
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Mustard, nitrogen (HN-1) [bis(2-chloroethyl)ethylamine]
Mustard, nitrogen (HN-2) [2,2'-dichloro-N-
methyldiethylamine N,N-bis(2-chloroethyl)methylamine]
Mustard, nitrogen (HN-3) [tris(2-chloroethyl)amine]
Organophosphate Pesticides, NOS1
Parathion
Phenol
Phosphamidon
R-33 (VR) [methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-methylpropyl ester]
Semivolatile Organic Compounds, NOS1
Tear gas (CS) [chlorobenzylidene malonitrile]
Tetramethylenedisulfotetramine
samples for the contaminants identified below and
ppropriate determinative method for these analytes.
CASRN
56073-10-0
28772-56-7
470-90-6
96-24-2
1445-76-7
7040-57-5
2921-88-2
329-99-7
NA
82-66-6
139-87-7
22224-92-6
437-38-7
10265-92-6
60-34-4
105-59-9
1189-87-3
538-07-8
51-75-2
555-77-1
NA
56-38-2
108-95-2
13171-21-6
159939-87-4
NA
2698-41-1
80-12-6
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Section 5 - Selected Chemical Methods
Analyte(s)
Triethanolamine (TEA)
VE [phosphonothioic acid, ethyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VG [phosphonothioic acid, S-(2-(diethylamino)ethyl)
O,O-diethyl ester]
VM [phosphonothioic acid, methyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
CASRN
102-71-6
21738-25-0
78-53-5
21770-86-5
1 NOS = Not otherwise specified
Approximately 10 g of solid sample is mixed with an equal amount of anhydrous sodium sulfate and
placed in an extraction thimble or between two plugs of glass wool. After adding the appropriate
surrogate amount, the sample is extracted using an appropriate solvent in an automated Soxhlet extractor.
The extract is dried with sodium sulfate to remove residual moisture, concentrated and exchanged, as
necessary, into a solvent compatible with the cleanup or determinative procedure for analysis.
Source: EPA Method 3541 (SW-846): Automated Soxhlet Extraction, Revision 0, 1994.
^^
5.2.20 EPA Method 3545A (SW-846): Pressurized Fluid Extraction (PFE)
This method should be used for preparation of solid samples for the contaminants identified below and
listed in Appendix A. Refer to Appendix A for the appropriate determinative method for these analytes.
Analyte(s)
Brodifacoum
Bromadiolone
Chlorfenvinphos
3-Chloro-1 ,2-propanediol
Chloropicrin
Chlorosarin
Chlorosoman
Chlorpyrifos
Crimidine
Cyclohexyl sarin (GF)
Dichlorvos
Dicrotophos
Diesel Range Organics
Diisopropyl methylphosphonate (DIMP)
Dimethylphosphite
CASRN
56073-10-0
28772-56-7
470-90-6
96-24-2
76-06-2
1445-76-7
7040-57-5
2921-88-2
535-89-7
329-99-7
62-73-7
141-66-2
NA
1445-75-6
868-85-9
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Section 5 - Selected Chemical Methods
Analyte(s)
Dimethylphosphoramidic acid
Diphacinone
1,4-Dithiane
EA2192 [Diisopropylaminoethyl
methylthiolophosphonate]
Ethyl methylphosphonic acid (EMPA)
Ethyldichloroarsine (ED)
N-Ethyldiethanolamine (EDEA)
Fenamiphos
Fentanyl
Isopropyl methylphosphonic acid (IMPA)
Methamidophos
Methyl hydrazine
Methyl parathion
N-Methyldiethanolamine (MDEA)
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Methylphosphonic acid (MPA)
Mevinphos
Mustard, nitrogen (HN-1) [bis(2-chloroethyl)ethylamine]
Mustard, nitrogen (HN-2) [2,2'-dichloro-N-
methyldiethylamine N,N-bis(2-chloroethyl)methylamine]
Mustard, nitrogen (HN-3) [tris(2-chloroethyl)amine]
Nicotine sulfate
Organophosphate Pesticides, NOS1
Parathion
Phencyclidine
Phenol
Phorate
Phosphamidon
Pinacolyl methyl phosphonic acid (PMPA)
R-33 (VR) [methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-methylpropyl ester]
CASRN
33876-51-6
82-66-6
505-29-3
73207-98-4
1832-53-7
598-14-1
139-87-7
22224-92-6
437-38-7
1832-54-8
10265-92-6
60-34-4
298-00-0
105-59-9
1189-87-3
993-13-5
7786-34-7
538-07-8
51-75-2
555-77-1
54-11-5
NA
56-38-2
77-10-1
108-95-2
298-02-2
13171-21-6
616-52-4
159939-87-4
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Section 5 - Selected Chemical Methods
Analyte(s)
Semivolatile Organic Compounds, NOS1
Soman (GD)
Strychnine
Tabun (GA)
Tear gas (CS) [chlorobenzylidene malonitrile]
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Thiodiglycol (TDG)
1,4-Thioxane
Triethanolamine (TEA)
Trimethyl phosphite
VE [phosphonothioic acid, ethyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VG [phosphonothioic acid, S-(2-(diethylamino)ethyl)
O,O-diethyl ester]
VM [phosphonothioic acid, methyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
CASRN
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
111-48-8
15980-15-1
102-71-6
121-45-9
21738-25-0
78-53-5
21770-86-5
1 NOS = Not otherwise specified
Approximately 10 to 30 g of soil sample is prepared for extraction either by air drying the sample, or by
mixing the sample with anhydrous sodium sulfate or pelletized diatomaceous earth. The sample is then
ground and loaded into the extraction cell. The extraction cell containing the sample is heated to the
extraction temperature, pressurized with the appropriate solvent system, and extracted for 5 minutes (or as
recommended by the instrument manufacturer). The extract may be concentrated, if necessary, and
exchanged into a solvent compatible with the cleanup or determinative step being employed. This
method has been validated for solid matrices containing 250 to 12,500 ug/kg of semivolatile organic
compounds, 250 to 2500 ug/kg of organophosphorus pesticides, 5 to 250 ug/kg of organochlorine
pesticides, 50 to 5000 ug/kg of chlorinated herbicides, 1 to 1400 ug/kg of PCBs, and 1 to 2500 ng/kg of
poly chlorinated dibenzo-^-dioxins/poly chlorinated dibenzofurans (PCDDs/PCDFs).
Please note: Sodium sulfate can cause clogging, and air drying or diatomaceous earth may be preferred.
Source: EPA Method 3545A (SW-846): Pressurized Fluid Extraction (PFE), Revision 1, 1998.
http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3545a.pdf
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Section 5 - Selected Chemical Methods
5.2.21 EPA Method 3571 (SW-846): Extraction of Solid and Aqueous Samples for
Chemical Agents
This method should be used for preparation of solid, organic solid, nonaqueous liquid, aqueous liquid
and drinking water samples for the contaminant identified below and listed in Appendix A. Refer to
Appendix A for the appropriate determinative method for these analytes.
Analyte(s)
Mustard, sulfur/ Mustard gas (HD)_
Sarin (GB)
VX
CASRN
505-60-2
107-44-8
50782-69-9
This method provides procedures for sample collection and extraction of the referenced compounds from
solids and aqueous samples. A separate extract is required for each agent to be measured. Glacial acetic
acid is added as a preservative to samples being assayed for GB and glacial acetic acid/sodium chloride is
a preservative for samples assayed for HD. No preservative is added for VX. Samples are extracted with
10% isopropanol in dichloromethane by vortex mixing and filtered, if necessary. An optional water wash
is included for VX that back-extracts the compound from heavy organics that could interfere with the
assay. An optional column cleanup procedure is described to separate GB from heavy organics, if
needed. Solvents are used to elute the extract first through the Carboprep90 column, then the silica
column.
Source: EPA Method 3571: Extraction of Solid and Aqueous Samples for Chemical Agents.
http://www.cpa.gov/cpaoswcr/hazwastc/tcst/pdfs/3571.pdf
5.2.22 EPA Method 3580A (SW-846): Waste Dilution
This method should be used for preparation of non-aqueous liquid/organic solid samples for the
contaminants identified below and listed in Appendix A. Refer to Appendix A for the appropriate
determinative method for these analytes.
Analyte(s)
Brodifacoum
Bromadiolone
Chlorfenvinphos
3-Chloro-1 ,2-propanediol
Chloropicrin
Chlorosarin
Chlorosoman
Chlorpyrifos
Crimidine
Cyclohexyl sarin (GF)
Dichlorvos
Dicrotophos
CASRN
56073-10-0
28772-56-7
470-90-6
96-24-2
76-06-2
1445-76-7
7040-57-5
2921-88-2
535-89-7
329-99-7
62-73-7
141-66-2
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Section 5 - Selected Chemical Methods
Analyte(s)
Diesel Range Organics
Diisopropyl methylphosphonate (DIMP)
Dimethylphosphite
Dimethylphosphoramidic acid
Diphacinone
1,4-Dithiane
EA2192 [Diisopropylaminoethyl
methylthiolophosphonate]
Ethyl methylphosphonic acid (EMPA)
Ethyldichloroarsine (ED)
N-Ethyldiethanolamine (EDEA)
Fenamiphos
Fentanyl
Isopropyl methylphosphonic acid (IMPA)
Methamidophos
Methyl hydrazine
Methyl parathion
N-Methyldiethanolamine (MDEA)
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Methylphosphonic acid (MPA)
Mevinphos
Mustard, nitrogen (HN-1) [bis(2-chloroethyl)ethylamine]
Mustard, nitrogen (HN-2) [2,2'-dichloro-N-
methyldiethylamine N,N-bis(2-chloroethyl)methylamine]
Mustard, nitrogen (HN-3) [tris(2-chloroethyl)amine]
Nicotine sulfate
Organophosphate Pesticides, NOS1
Parathion
Phencyclidine
Phenol
Phorate
Phosphamidon
CASRN
NA
1445-75-6
868-85-9
33876-51-6
82-66-6
505-29-3
73207-98-4
1832-53-7
598-14-1
139-87-7
22224-92-6
437-38-7
1832-54-8
10265-92-6
60-34-4
298-00-0
105-59-9
1189-87-3
993-13-5
7786-34-7
538-07-8
51-75-2
555-77-1
54-11-5
NA
56-38-2
77-10-1
108-95-2
298-02-2
13171-21-6
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Section 5 - Selected Chemical Methods
Analyte(s)
Pinacolyl methyl phosphonic acid (PMPA)
R-33 (VR) [methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-methylpropyl ester]
Semivolatile Organic Compounds, NOS1
Soman (GD)
Strychnine
Tabun (GA)
Tear gas (CS) [chlorobenzylidene malonitrile]
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Thiodiglycol (TDG)
1,4-Thioxane
Triethanolamine (TEA)
Trimethyl phosphite
VE [phosphonothioic acid, ethyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VG [phosphonothioic acid, S-(2-(diethylamino)ethyl)
O,O-diethyl ester]
VM [phosphonothioic acid, methyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
CASRN
616-52-4
159939-87-4
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
111-48-8
15980-15-1
102-71-6
121-45-9
21738-25-0
78-53-5
21770-86-5
1 NOS = Not otherwise specified
This method describes solvent dilution of a non-aqueous waste sample prior to cleanup and/or analysis.
One gram of sample is weighed into a capped tube and the sample is diluted to 10.0 mL with an
appropriate solvent. The method is designed for wastes that may contain organic chemicals at a
concentration greater than 20,000 mg/kg and that are soluble in the dilution solvent.
Source: EPA Method 3580A (SW-846): Waste Dilution, Revision 1, 1992.
http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3580a.pdf
5.2.23 EPA Method 3585 (SW-846): Waste Dilution for Volatile Organics
This method should be used for preparation of non-aqueous liquid/organic solid samples for the
contaminants identified below and listed in Appendix A. Refer to Appendix A for the appropriate
determinative method for these analytes.
Analyte(s)
Allyl alcohol
Carbon disulfide
2-Chloroethanol
CASRN
107-18-6
75-15-0
107-07-3
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Section 5 - Selected Chemical Methods
Analyte(s)
Cyanogen chloride
1,2-Dichloroethane
Ethylene oxide
Gasoline Range Organics
Kerosene
Propylene oxide
CASRN
506-77-4
107-06-2
75-21-8
NA
64742-81-0
75-56-9
The following analytes should be prepared by this method (and determined by the corresponding SW-846 Method
8260C) only if problems (e.g., insufficient recovery, interferences) occur when using the sample
preparation/determinative techniques identified for these analytes in Appendix A.
1,4-Dithiane
1,4-Thioxane
505-29-3
15980-15-1
This method describes solvent dilution of a non-aqueous waste sample prior to direct injection analysis.
It is designed for use in conjunction with GC or GC-MS analysis of wastes that may contain organic
chemicals at a concentration greater than 1 mg/kg and that are soluble in the dilution solvent. Highly
contaminated or highly complex samples may be diluted prior to analysis for volatiles using direct
injection. One gram of sample is weighed into a capped tube or volumetric flask. The sample is diluted
to 2.0 to 10.0 mL with «-hexadecane or other appropriate solvent. Diluted samples are injected into the
GC or GC-MS for analysis.
Source: EPA Method 3585 (SW-846): Waste Dilution for Volatile Organics, Revision 0, 1996.
http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3585.pdf
5.2.24 EPA Method 5030C (SW-846): Purge-and-Trap for Aqueous Samples
This method should be used for preparation of aqueous liquid and/or drinking water samples for the
contaminants identified below and listed in Appendix A. Note: For carbon disulfide, 1,2-dichloroethane,
and volatile organic compounds only, EPA Method 524.2 (rather than Method 5030C) should be used for
preparation of drinking water samples. Refer to Appendix A for the appropriate determinative method for
these analytes.
Analyte(s)
Allyl alcohol
Carbon disulfide
2-Chloroethanol
Cyanogen chloride
1,2-Dichloroethane
Ethylene oxide
Gasoline Range Organics
Kerosene
Propylene oxide
CASRN
107-18-6
75-15-0
107-07-3
506-77-4
107-06-2
75-21-8
NA
64742-81-0
75-56-9
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Analyte(s)
CASRN
The following analytes should be prepared by this method (and determined by the corresponding SW-846 Method
8260C) only if problems (e.g., insufficient recovery, interferences) occur when using the sample
preparation/determinative techniques identified for these analytes in Appendix A.
1,4-Dithiane
1,4-Thioxane
505-29-3
15980-15-1
This method describes a purge-and-trap procedure for the analysis of volatile organic compounds (VOCs)
in aqueous liquid samples and water miscible liquid samples. An inert gas is bubbled through a portion of
the aqueous liquid sample at ambient temperature, and the volatile components are transferred from the
aqueous liquid phase to the vapor phase. The vapor is swept through a sorbent column where the volatile
components are adsorbed. After purging is completed, the sorbent column is heated and backflushed with
inert gas to desorb the components onto a gas chromatographic column.
Source: EPA Method 5030C (SW-846): Purge-and-Trap for Aqueous Samples, Revision 3, 2003.
_^
5.2.25 EPA Method 5035A (SW-846): Closed-System Purge-and-Trap and Extraction for
Volatile Organics in Soil and Waste Samples
This method should be used for preparation of solid samples for the contaminants identified below and
listed in Appendix A. Refer to Appendix A for the appropriate determinative method for these analytes.
Analyte(s)
Allyl alcohol
Carbon disulfide
2-Chloroethanol
Cyanogen chloride
1,2-Dichloroethane
Ethylene oxide
Gasoline Range Organics
Kerosene
Propylene oxide
CASRN
107-18-6
75-15-0
107-07-3
506-77-4
107-06-2
75-21-8
NA
64742-81-0
75-56-9
The following analytes should be prepared by this method (and determined by the corresponding SW-846 Method
8260C) only if problems (e.g., insufficient recovery, interferences) occur when using the sample
preparation/determinative techniques identified for these analytes in Appendix A.
1,4-Dithiane
1,4-Thioxane
505-29-3
15980-15-1
This method describes a closed-system purge-and-trap process for analysis of volatile organic compounds
(VOCs) in solid samples containing low levels (0.5 to 200 ug/kg) of VOCs. The method also provides
specific procedures for preparation of samples containing high levels (>200 ug/kg ) of VOCs. For low-
level VOCs, a 5-g sample is collected into a vial that is placed into an autosampler device. Reagent
water, surrogates, and internal standards are added automatically, and the vial is heated to 40 °C. The
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volatiles are purged into an appropriate trap using an inert gas combined with sample agitation. When
purging is complete, the trap is heated and backflushed with helium to desorb the trapped sample
components into a gas chromatograph for analysis. For high-level VOCs, samples are either collected
into a vial that contains a water-miscible organic solvent or a portion of sample is removed from the vial
and dispersed in a water-miscible solvent. An aliquot of the solvent is added to reagent water, along with
surrogates and internal standards, then purged and analyzed using an appropriate determinative method
(e.g., Method 8015C or 8260C (SW-846)).
Source: EPA Method 5035A (SW-846): Closed-System Purge-and-Trap and Extraction for Volatile
Organics in Soil and Waste Samples, Draft Revision 1, 2002.
http://www.cpa.gov/cpaoswcr/liazwasto/tost/pdfs/5035a rl.pdf
5.2.26 EPA Method 6010C (SW-846): Inductively Coupled Plasma - Atomic Emission
Spectrometry
This method should be used for analysis of solid and non-aqueous liquid/organic solid samples for the
contaminants identified below and listed in Appendix A. Appropriate sample preparation techniques
should be used prior to analysis (refer to Appendix A). Note: Osmium tetroxide and titanium
tetrachloride are not of concern in non-aqueous liquid/organic solid samples.
Analyte(s)
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
2-Chlorovinylarsonous acid (CVAA)
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]1
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite Oxide
Osmium tetroxide
Sodium arsenite
Thallium sulfate
Titanium tetrachloride
Vanadium pentoxide
CASRN
7803-55-6
740-38-2
1327-53-3
85090-33-1
541-25-3
40334-69-8
40334-70-1
1306-02-1
20816-12-0
7784-46-5
10031-59-1
7550-45-0
1314-62-1
1 Laboratory testing is currently underway for speciation of lewisite 1 using GC-MS techniques.
This method determines arsenic trioxide, lewisite, lewisite degradation products, and sodium arsenite as
total arsenic; osmium tetroxide as osmium; thallium sulfate as thallium; titanium tetrachloride as titanium;
and ammonium metavanadate and vanadium pentoxide as total vanadium. Soil samples (prepared using
SW-846 Methods 3050B), and non-aqueous liquid/organic solid samples (prepared using SW-846
Methods 3031) are analyzed by Inductively Coupled Plasma - Atomic Emission Spectrometry (ICP-
AES). Detection limits vary with each analyte. Estimated instrument detection limits for arsenic and
titanium are 30 (ig/L and 5.0 (ig/L, respectively. The upper end of the analytical range may be extended
by sample dilution.
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Source: EPA Method 6010C (SW-846): Inductively Coupled Plasma-Atomic Emission Spectrometry,
Revision 3, 2000. http://www.cpa.gov/cpaoswcr/hazwastc/tcst/pdfs/6010c.pdf
5.2.27 EPA Method 6020A (SW-846): Inductively Coupled Plasma - Mass Spectrometry
This method should be used for analysis of solid and non-aqueous liquid/organic solid samples for the
contaminants identified below and listed in Appendix A. Appropriate sample preparation techniques
should be used prior to analysis (refer to Appendix A). Note: Titanium tetrachloride is not of concern in
non-aqueous liquid/organic solid samples.
Analyte(s)
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
2-Chlorovinylarsonous acid (CVAA)
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]1
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite oxide
Sodium arsenite
Thallium sulfate
Titanium tetrachloride
Vanadium pentoxide
CASRN
7803-55-6
740-38-2
1327-53-3
85090-33-1
541-25-3
40334-69-8
40334-70-1
1306-02-1
7784-46-5
10031-59-1
7550-45-0
1314-62-1
1 Laboratory testing is currently underway for speciation of lewisite 1 using GC-MS techniques.
This method will determine arsenic trioxide, lewisite, lewisite degradation products, and sodium arsenite
as total arsenic. The method also will determine thallium sulfate as total thallium, titanium tetrachloride
as titanium, and ammonium metavanadate and vanadium pentoxide as total vanadium. Aqueous samples
(prepared using SW-846 Method 5050), soil samples (prepared using SW-846 Methods 3050B or 5050),
non-aqueous liquid/organic solid samples (prepared using SW-846 Methods 3050B or 3031), and air
filter/particle samples (prepared using IO Method 3.5) are analyzed by Inductively Coupled Plasma -
Mass Spectrometry. Instrument detection limits, sensitivities, and linear ranges vary with sample type,
instrumentation, and operation conditions. In relatively simple sample types, detection limits will
generally be below 0.1 (ig/L. Less sensitive elements, such as arsenic, may have detection limits of 1.0
(ig/L or higher. The upper end of the analytical range may be extended by sample dilution.
Source: EPA Method 6020A (SW-846): Inductively Coupled Plasma-Mass Spectrometry, Revision 1,
1998. http://www.epa.gov/epaoswer/hazwaste/test/pdfs/6020a.pdf
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5.2.28 EPA Method 7010 (SW-846): Graphite Furnace Atomic Absorption
Spectrophotometry
This method should be used for analysis of solid samples for the contaminant identified below and listed
in Appendix A. Appropriate sample preparation techniques should be used prior to analysis (refer to
Appendix A).
Analyte(s)
Arsine
CASRN
7784-42-1
This method determines arsine as arsenic in environmental samples. Soil samples (prepared using SW-
846 Method 3 05 OB) are analyzed by Graphite Furnace Atomic Absorption Spectrophotometry (GFAA).
A representative aliquot of the sample is placed in the graphite tube in the furnace, evaporated to dryness,
charred, and atomized. Detection limits vary with each sample type and instrument used. The analytical
range may be extended by sample dilution.
Source: EPA Method 7010 (SW-846): Graphite Furnace Atomic Absorption Spectrophotometry,
Revision 0, 1998.
5.2.29 EPA Method 7470A (SW-846): Mercury in Liquid Wastes (Manual Cold-Vapor
Technique)
This method is not currently listed for any of the analyte/sample type combinations included in Appendix
A. If problems occur when using EPA SW-846 Method 7473, then this method should be used for
preparation and analysis of aqueous liquid samples for the contaminant identified below and listed in
Appendix A. (See Footnote 12 of Appendix A.)
Analyte(s)
Mercury, Total
Methoxyethylmercuric acetate
CASRN
7439-97-6
151-38-2
A 100-mL aqueous sample is digested with acids, permanganate solution, persulfate solution, and heat.
The sample is cooled and reduced with hydroxylamine-sodium chloride solution. Just prior to analysis,
the sample is treated with Sn(II), reducing the mercury to Hg(0). The reduced sample is sparged and the
mercury vapor is analyzed by cold vapor atomic absorption. The detection limit for the method is less
than 0.2 (ig/L. Chloride and copper are potential interferences.
Source: EPA Method 7470A (SW-846): Mercury in Liquid Waste (Manual Cold-Vapor Technique),
Revision 1, 1994. iMjsi/lwwy^^^
5.2.30 EPA Method 7471 B (SW-846): Mercury in Solid or Semisolid Wastes (Manual Cold-
Vapor Technique)
This method is not currently listed for any of the analyte/sample type combinations included in Appendix
A. If problems occur when using EPA SW-846 Method 7473, then this method should be used for
preparation and analysis of solid samples for the contaminant identified below and listed in Appendix
A. (See Footnote 12 of Appendix A.)
Analyte(s)
Mercury, Total
Methoxyethylmercuric acetate
CASRN
7439-97-6
151-38-2
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A 0.5-g to 0.6-g sample is digested with aqua regia, permanganate solution, and heat. The sample is
cooled and reduced with hydroxylamine-sodium chloride solution. Just prior to analysis, the sample is
treated with Sn(II), reducing the mercury to Hg(0). The reduced sample is sparged and the mercury vapor
is analyzed by cold vapor atomic absorption. Chloride and copper are potential interferences.
Source: EPA Method 747IB (SW-846): Mercury in Solid or Semisolid Waste (Manual Cold-Vapor
Technique), Revision 2, 1998. http://www.cpa.gov/cpaoswcr/liazwastc/tcst/pdfs/7471b.pdf
5.2.31 EPA Method 7473 (SW-846): Mercury in Solids and Solutions by Thermal
Decomposition, Amalgamation, and Atomic Absorption Spectrophotometry
This method should be used for preparation and analysis of solid and aqueous liquid samples for the
contaminants identified below and listed in Appendix A.
Analyte(s)
Mercury, Total1
Methoxyethylmercuric acetate1
CASRN
7439-97-6
151-38-2
1 If equipment is not available, use CVAA Methods 7471B (EPA SW-846) for solid samples and 7470A (EPA SW-
846) for aqueous liquid samples.
Controlled heating in an oxygenated decomposition furnace is used to liberate mercury from solid and
aqueous samples. The sample is dried and then thermally and chemically decomposed within the furnace.
The decomposition products are carried by flowing oxygen to the catalytic section of the furnace, where
oxidation is completed and halogens and nitrogen/sulfur oxides are trapped. The remaining
decomposition products are then carried to an amalgamator that selectively traps mercury. After the
system is flushed with oxygen to remove any remaining gases or decomposition products, the
amalgamator is rapidly heated, releasing mercury vapor. Flowing oxygen carries the mercury vapor
through absorbance cells positioned in the light path of a single wavelength atomic absorption
spectrophotometer. Absorbance (peak height or peak area) is measured at 253.7 nm as a function of
mercury concentration. The typical working range for this method is 0.05 - 600 ng. The instrument
detection limit (IDL) is 0.01-ng total mercury.
Source: EPA Method 7473 (SW-846): Mercury in Solids and Solutions by Thermal Decomposition,
Amalgamation, and Atomic Absorption Spectrophotometry, Revision 0, 1998.
http://www.epa.gov/epaoswer/hazwaste/test/pdfs/7473.pdf
5.2.32 EPA Method 7580 (SW-846): White Phosphorus by Solvent Extraction and Gas
Chromatography (GC)
This method should be used for preparation and analysis of solid, non-aqueous liquid/organic solid,
aqueous liquid, and drinking water samples for the contaminant identified below and listed in Appendix
A.
Analyte(s)
White Phosphorus
CASRN
12185-10-3
Method 7580 may be used to determine the concentration of white phosphorus in soil, sediment, and
water samples using solvent extraction and gas chromatography (GC). Water samples are extracted by
one of two procedures, depending on the sensitivity required. For the more sensitive procedure, a 500-
mL water sample is extracted with 50 mL of diethyl ether. The extract is concentrated by back extraction
with reagent water, yielding a final extract volume of approximately 1.0 mL. A 1.0 (iL aliquot of this
extract is injected into a GC equipped with a nitrogen-phosphorus detector (NPD). This procedure
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Section 5 - Selected Chemical Methods
provides sensitivity on the order of 0.01 (ig/L. Wet soil or sediment samples are analyzed by extracting a
40 g wet-weight aliquot of the sample with a mixture of 10.0 mL degassed reagent water and 10.0 mL
isooctane. The extraction is performed in a glass jar on a platform shaker for 18 hours. A 1 .0 (iL aliquot
of the extract is analyzed by GC-NPD, providing sensitivity on the order of 1
Source: EPA Method 7580 (SW-846): White Phosphorus (P4) by Solvent Extraction and Gas
Chromatography, Revision 0, 1996. httg7/www1cga.goy/sw^84_6/Bdfe/7580.pdf
5.2.33 EPA Method 8015C (SW-846): Nonhalogenated Organics Using GC-FID
This method should be used for analysis of solid, non-aqueous liquid/organic solid, aqueous liquid, and
drinking water samples for the contaminants identified below and listed in Appendix A. Appropriate
sample preparation techniques should be used prior to analysis (refer to Appendix A).
Analyte(s)
Diesel Range Organics
Gasoline Range Organics
Kerosene
CASRN
NA
NA
64742-81-0
This method provides gas chromatographic (GC) conditions for the detection of certain nonhalogenated
volatile and semivolatile organic compounds. Depending on the analytes of interest, samples may be
introduced into the GC by a variety of techniques including purge-and-trap, direct injection of aqueous
liquid samples, and solvent extraction. An appropriate column and temperature program are used in the
gas chromatograph to separate the organic compounds. Detection is achieved by a flame ionization
detector (FID). The method allows the use of packed or capillary columns for the analysis and
confirmation of the non-halogenated individual analytes. The estimated method detection limits vary
with each analyte and range between 2 and 48 ug/L for aqueous liquid samples. The method detection
limits in other matrices have not been evaluated for this method. The analytical range depends on the
target analyte(s) and the instrument used.
Source: EPA Method 8015C (SW-846): Nonhalogenated Organics Using GC/FID, Revision 3, 2000.
http://www.cpa.gov/cpaoswcr/liazwasto/tost/pdfs/8015c.pdf
5.2.34 EPA Method 8260C (SW-846): Volatile Organic Compounds by Gas
Chromatography-Mass Spectrometry (GC-MS)
This method should be used for analysis of solid, non-aqueous liquid/organic solid, aqueous liquid, and
drinking water samples for the contaminants identified below and listed in Appendix A. Appropriate
sample preparation techniques should be used prior to analysis (refer to Appendix A). Note: For carbon
disulfide and 1,2-dichloroethane only, EPA Method 524.2 (rather than 8260C) should be used for analysis
of drinking water samples.
Analyte(s)
Allyl alcohol
Carbon disulfide
2-Chloroethanol
Cyanogen chloride
1,2-Dichloroethane
CASRN
107-18-6
75-15-0
107-07-3
506-77-4
107-06-2
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Analyte(s)
Ethylene oxide
Propylene oxide
CASRN
75-21-8
75-56-9
The following analytes should be determined by this method (and corresponding sample preparation methods)
only if problems (e.g., insufficient recovery, interferences) occur when using the sample preparation/determinative
techniques identified for these analytes in Appendix A.
1,4-Dithiane
1,4-Thioxane
505-29-3
15980-15-1
Volatile compounds are introduced into a gas chromatograph (GC) by purge-and-trap or other procedures
(see Section 1.2 in Method 8260C). The analytes can be introduced directly to a wide-bore capillary
column or cryofocused on a capillary pre-column before being flash evaporated to a narrow-bore capillary
for analysis. Alternatively, the effluent from the trap is sent to an injection port operating in the split
mode for injection to a narrow-bore capillary column. The column is temperature-programmed to
separate the analytes, which are then detected with a mass spectrometer (MS) interfaced to the GC.
Analytes eluted from the capillary column are introduced into the mass spectrometer via a jet separator or
a direct connection. The estimated quantitation limit (EQL) for an individual analyte is dependent on the
instrument as well as the choice of sample preparation/introduction method. Using standard quadrupole
instrumentation and the purge-and-trap technique, estimated quantitation limits are 5 ug/kg (wet weight)
for soil/sediment samples and 5 ug/L for ground water. Somewhat lower limits may be achieved using an
ion trap mass spectrometer or other instrumentation of improved design. No matter which instrument is
used, EQLs will be proportionately higher for sample extracts and samples that require dilution or when a
reduced sample size is used to avoid saturation of the detector.
Source: EPA Method 8260C (SW-846): Volatile Organic Compounds by Gas Chromatography-Mass
Spectrometry (GC-MS), Revision 3, 2006. http://www.epa.gov/epaoswer/hazwaste/test/pdfs/8260c.pdf
5.2.35 EPA Method 8270D (SW-846): Semivolatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC-MS)
This method should be used for analysis of solid, non-aqueous liquid/organic solid, aqueous liquid, and
drinking water samples for the contaminants identified below and listed in Appendix A. Appropriate
sample preparation techniques should be used prior to analysis (refer to Appendix A). Note: For
dichlorvos, fenamiphos, mevinphos, and semivolatile organic compounds only, EPA Method 525.2
(rather than Method 8270D) should be used for analysis of drinking water samples. For organophosphate
pesticides only, EPA Methods 614 and 507 should be used for analysis of aqueous liquid and drinking
water samples, respectively. For chloropicrin only, EPA Method 551.1 should be used for analysis of
aqueous liquid and drinking water samples.
Analyte(s)
Chlorfenvinphos
3-Chloro-1 ,2-propanediol1
Chloropicrin2
Chlorosarin
Chlorosoman
Chlorpyrifos
Crimidine3
CASRN
470-90-6
96-24-2
76-06-2
1445-76-7
7040-57-5
2921-88-2
535-89-7
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Analyte(s)
Cyclohexyl sarin (GF)
Dichlorvos
Dicrotophos
Diisopropyl methylphosphonate (DIMP)
Dimethylphosphite
1,4-Dithiane4
Ethyldichloroarsine (ED)
Fenamiphos
Methamidophos
Methyl hydrazine
Methyl parathion
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Mevinphos
Mustard, nitrogen (HN-1) [bis(2-chloroethyl)ethylamine]
Mustard, nitrogen (HN-2) [2,2'-dichloro-N-
methyldiethylamine N,N-bis(2-chloroethyl)methylamine]
Mustard, nitrogen (HN-3) [tris(2-chloroethyl)amine]
Mustard, sulfur/ Mustard gas (HD)5
Nicotine sulfate
Organophosphate Pesticides, NOS6
Parathion
Phencyclidine
Phenol
Phorate
Phosphamidon
R-33 (VR) [methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-methylpropyl ester]
Sarin (GB)5
Semivolatile Organic Compounds, NOS6
Soman (GD)
Strychnine
Tabun (GA)
CASRN
329-99-7
62-73-7
141-66-2
1445-75-6
868-85-9
505-29-3
598-14-1
22224-92-6
10265-92-6
60-34-4
298-00-0
1189-87-3
7786-34-7
538-07-8
51-75-2
555-77-1
505-60-2
54-11-5
NA
56-38-2
77-10-1
108-95-2
298-02-2
13171-21-6
159939-87-4
107-44-8
NA
96-64-0
57-24-9
77-81-6
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Analyte(s)
Tear gas (CS) [chlorobenzylidene malonitrile]
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine2'7'8
Thiodiglycol (TDG)
1,4-Thioxane4
Trimethyl phosphite
VE [phosphonothioic acid, ethyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VG [phosphonothioic acid, S-(2-(diethylamino)ethyl)
O,O-diethyl ester]
VM [phosphonothioic acid, methyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VX [O-ethyl-S-(2-diisopropylaminoethyl)methyl-
phosphonothiolate]5
CASRN
2698-41-1
107-49-3
80-12-6
111-48-8
15980-15-1
121-45-9
21738-25-0
78-53-5
21770-86-5
50782-69-9
The following analyte should be determined by this method only if LC-MS (electrospray) procedures are not
available to the laboratory. These analytes should be analyzed with GC-MS procedures using derivatization based
on SW-846 Method 8270D. Sample preparation methods should remain the same. Both electrospray LC-MS and
GC/MS derivatization procedures are currently under development.
Dimethylphosphoramidic acid
EA2192 [Diisopropylaminoethyl
methylthiolophosphonate]
Ethylmethyl phosphonate (EMPA)
Isopropyl methylphosphonic acid (IMPA)
Methylphosphonic acid (MPA)
Pinacolyl methyl phosphonic acid (PMPA)
33876-51-6
73207-98-4
1832-53-7
1832-54-8
993-13-5
616-52-4
1 For this analyte, SW-846 Method 8270D must be modified to include a derivatization step.
2 This analyte requires determination using an injection port temperature of less than 200°C.
3 If problems occur when using this method, it is recommended that SW-846 Method 8321B be used.
4 If problems occur when using this method, it is recommended that SW-846 Method 8260C and appropriate
corresponding sample preparation procedures (i.e., 5035A for solid samples, 3585 for non-aqueous liquid/organic
solid samples, and 5030C for aqueous liquid and drinking water samples) be used.
5 For this analyte, refer to EPA SW-846 Method 8271 for GC-MS conditions
6 NOS = Not otherwise specified
7 This analyte may require SIM analyses in order to be determined.
8 When analyzing fortetramine, the injection temperature must not exceed 250°C (the decomposition temperature of
tetramine).
Samples are prepared for analysis by gas chromatography/mass spectrometry using the appropriate
sample preparation and, if necessary, sample cleanup procedures. Semivolatile compounds are
introduced into the GC-MS by injecting the sample extract into a gas chromatograph (GC) with a narrow-
bore fused-silica capillary column. The GC column is temperature-programmed to separate the analytes,
which are then detected with a mass spectrometer (MS) connected to the GC. Analytes eluted from the
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capillary column are introduced into the MS. For the determination of 3-chloro-l,2-propanediol,
dimethylphosphoramidic acid, EA2192, EMPA, IMPA, MPA, and pinacolyl methyl phosphonic acid, a
derivatization step is required prior to injection into the GC-MS. The phosphonic acids require
derivatization with a trimethylsilyl agent and 3-chloro-l,2-propanediol requires derivatization with a
heptafluorobutyryl agent. The estimated detection limits vary with each analyte and range between 10
and 1000 (ig/L for aqueous liquid samples and 660 and 3300 (ig/kg for soil samples. The analytical range
depends on the target analyte(s) and the instrument used.
Source: EPA Method 8270D (SW-846): Semivolatile Organic Compounds by Gas Chromatography/Mass
Spectrometry (GC/MS), Revision 4, 1998. http://www.cpa.gov/cpaoswcr/hazwastc/tcst/pdfs/8270d.pdf
5.2.36 EPA Method 8315A (SW-846): Determination of Carbonyl Compounds by High
Performance Liquid Chromatography (HPLC)
This method should be used for preparation and analysis of solid, aqueous liquid, and drinking water
samples for the contaminant identified below and listed in Appendix A.
Analyte(s)
Formaldehyde
CASRN
50-00-0
A measured volume of aqueous liquid sample (approximately 100 mL), or an appropriate amount of
solids extract (approximately 25 g), is buffered to pH 3 and derivatized with 2,4-dinitrophenylhydrazine
(DNPH). Using the appropriate extraction technique, the derivatives are extracted using methylene
chloride and the extracts are exchanged with acetonitrile prior to HPLC analysis. HPLC conditions are
described permitting the separation and measurement of various carbonyl compounds in the extract by
absorbance detection at 360 nm. If formaldehyde is the only analyte of interest, the aqueous liquid
sample and/or solid sample extract should be buffered to pH 5.0 to minimize the formation of artifact
formaldehyde. The method detection limit for formaldehyde varies depending on sample conditions and
instrumentation, but is approximately 6.2 (ig/L for aqueous liquid samples.
Source: EPA Method 8315A (SW-846): Determination of Carbonyl Compounds by High Performance
Liquid Chromatography (HPLC), Revision 1, 1996.
5.2.37 EPA Method 8316 (SW-846): Acrylamide, Acrylonitrile and Acrolein by High
Performance Liquid Chromatography (HPLC)
This method should be used for preparation and/or analysis of solid, non-aqueous liquid/organic solid,
aqueous liquid, and drinking water samples for the contaminants identified below and listed in Appendix
A. Note: Solid and non-aqueous liquid/organic solid samples are extracted with water prior to analysis.
Analyte(s)
Acrylamide
Acrylonitrile
Methyl acrylonitrile
CASRN
79-06-1
107-13-1
126-98-7
Samples are analyzed by High Performance Liquid Chromatography (HPLC). A 200-(iL aliquot is
injected onto a Ci8 reverse-phase column, and compounds in the effluent are detected with an ultraviolet
(UV) detector. Solid and non-aqueous liquid/organic solid samples should be water extracted prior to
injection. Aqueous liquid and drinking water samples can be directly injected. Acrylamide has an MDL
of 10 (ig/L; acrylonitrile has an MDL of 20 (ig/L.
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Source: EPA Method 8316 (SW-846): Acrylamide, Acrylonitrile and Acrolein by High Performance
Liquid Chromatography (HPLC). http://www.cpa.gov/cpaoswcr/tiazwastc/tcst/pdfs/8316.pdf
5.2.38 EPA Method 8318A (SW-846): A/-Methylcarbamates by High Performance Liquid
Chromatography (HPLC)
This method should be used for preparation and analysis of solid, non-aqueous liquid/organic solid, and
aqueous liquid samples for the contaminants identified below and listed in Appendix A.
Analyte(s)
Aldicarb (Temik)
Carbofuran (Furadan)
Methomyl
Oxamyl
CASRN
116-06-3
1563-66-2
16752-77-5
23135-22-0
TV-methylcarbamates are extracted from aqueous liquid samples with methylene chloride, and from soils,
oily solid waste, and oils with acetonitrile. The extract solvent is exchanged to methanol/ethylene glycol,
and the extract is cleaned using a Qg cartridge, filtered, and eluted on a Ci8 analytical column. After
separation, the target analytes are hydrolyzed and derivatized post-column, then quantified
fluorometrically. The sensitivity of the method usually depends on the level of interferences present,
rather than on instrument conditions. Waste samples with a high level of extractable fluorescent
compounds are expected to yield significantly higher detection limits. The estimated method detection
limits vary with each analyte and range between 1.7 to 9.4 ug/L for aqueous liquid samples and 10 to 50
ug/kg for soil samples.
Source: EPA Method 8318A (SW-846): N-Methylcarbamates by High Performance Liquid
Chromatography (HPLC), Revision 1, 2000. http://www.cpa.gov/cpaoswcr/hazwastc/tcst/pdfs/8318a.pdf
5.2.39 EPA Method 8321B (SW-846): Solvent-Extractable Nonvolatile Compounds by
High Performance Liquid Chromatography-Thermospray-Mass Spectrometry
(HPLC-TS-MS) or Ultraviolet (UV) Detection
This method should be used for analysis of solid, non-aqueous liquid/organic solid, aqueous liquid, and
drinking water samples for the contaminant identified below and listed in Appendix A.
Analyte(s)
Brodifacoum
Bromadiolone
Dimethylphosphoramidic acid1
Diphacinone
EA2192 [Diisopropylaminoethyl
methylthiolophosphonate]1
Ethylmethyl phosphonate (EMPA)1
N-Ethyldiethanolamine (EDEA)
Fentanyl
Isopropyl methylphosphonic acid (IMPA)1
CASRN
56073-10-0
28772-56-7
33876-51-6
82-66-6
73207-98-4
1832-53-7
139-87-7
437-38-7
1832-54-8
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Analyte(s)
N-Methyldiethanolamine (MDEA)
Methylphosphonic acid (MPA)1
Pinacolyl methyl phosphonic acid (PMPA)1
Triethanolamine (TEA)
CASRN
105-59-9
993-13-5
616-52-4
102-71-6
The following analyte should be determined by this method only if problems (e.g., insufficient recovery,
interferences) occur when using SW-846 Method 8270D. Sample preparation methods should remain the same as
those listed in Appendix A.
Crimidine2
535-89-7
LC-MS (electrospray) procedures are preferred for these analytes; however, if this technique is not available to the
laboratory, GC-MS procedures using derivatization based on SW-846 Method 8270D may be used. Sample
preparation methods should remain the same. Both electrospray LC-MS and GC/MS derivatization procedures are
currently under development.
2 This analyte needs to be determined using a wavelength of 230 nm.
This method provides reversed-phase high performance liquid chromatographic (RP-HPLC), thermospray
(TSP) mass spectrometric (MS), and ultraviolet (UV) conditions for detection of the target analytes.
Sample extracts can be analyzed by direct injection into the thermospray or onto a liquid
chromatographic-thermospray interface. A gradient elution program is used to separate the compounds.
Primary analysis may be performed by UV detection; however, positive results should be confirmed by
TSP-MS. Quantitative analysis may be performed by either TSP-MS or UV detection, using either an
external or internal standard approach. TSP-MS detection may be performed in either a negative
ionization (discharge electrode) mode or a positive ionization mode, with a single quadrupole mass
spectrometer. The use of MS-MS techniques is an option. The analytical range and detection limits vary
depending on the target analyte and instrument used.
Source: EPA Method 832IB (SW-846): Solvent-Extractable Nonvolatile Compounds by High
Performance Liquid Chromatography-Thermospray-Mass Spectrometry (HPLC-TSP-MS) or Ultraviolet
(UV) Detection, Revision 2, 1998. http://www.epa.gov/epaoswer/hazwaste/test/pdfs/8321b.pdf
5.2.40 EPA Method 8330B (SW-846): Nitroaromatics and Nitramines by High Performance
Liquid Chromatography (HPLC)
This method should be used for preparation and/or analysis of solid, non-aqueous liquid/organic solid,
aqueous liquid, and drinking water samples for the contaminants identified below and listed in Appendix
A. Note: Aqueous liquid and drinking water samples are prepared using Methods 3535A or 8330B prior
to analysis.
Analyte(s)
2-Amino-4,6-dinitrotoluene (2-Am-DNT)
4-Amino-2,6-dinitrotoluene (4-Am-DNT)
4-Aminopyridine
3,5-Dinitroaniline (3,5-DNA)
1,3-Dinitrobenzene (1,3-DNB)
2,4-Dinitrotoluene (2,4-DNT)
CASRN
35572-78-2
19406-51-0
504-24-5
618-87-1
99-65-0
121-14-2
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Analyte(s)
2,6-Dinitrotoluene (2,6-DNT)
Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)
Hexamethylenetriperoxidediamine (HMTD)
Methyl-2,4,6-trinitrophenylnitramine (Tetryl)
Nitrobenzene (NB)
Nitroglycerin (NG)
2-Nitrotoluene (2-NT)
3-Nitrotoluene (3-NT)
4-Nitrotoluene (4-NT)
Octahydro-1 ,3,5,7-tetranitro-1 ,3,5,7-tetrazocine (HMX)
Pentaerythritol tetranitrate (PETN)
Triacetone triperoxide (TATP)
1 ,3,5-Trinitrobenzene (1 ,3,5-TNB)
2,4,6-Trinitrotoluene(2,4,6-TNT)
CASRN
606-20-2
121-82-4
283-66-9
479-45-8
98-95-3
55-63-0
88-72-2
99-08-1
99-99-0
2691-41-0
78-11-5
17088-37-8
99-35-4
118-96-7
This method is intended for the trace analysis of explosives and propellant residues by high performance
liquid chromatography (HPLC) using a dual wavelength ultraviolet (UV) detector in a water, soil, or
sediment matrix. All of the compounds listed in this method are either used in the manufacture of
explosives or propellants, or they are the degradation products of compounds used for that purpose.
Samples are prepared for analysis by HPLC-UV using the appropriate sample preparation technique
(solid phase extraction by 3535A or solvent extraction by 8330B) and, if necessary, sample cleanup
procedures. Method 8330 provides a salting-out extraction procedure for low concentration (ppt or ng/L)
of explosives residues in surface or ground water. Direct injection of diluted and filtered water samples
can be used for water samples of higher concentration. Soil and sediment samples are extracted using
acetonitrile in an ultrasonic bath, filtered and chromatographed. The detection limits, ranges, and
interferences depend on the target compound.
Source: EPA Method 8330B (SW-846): Nitroaromatics, Nitramines, and Nitrate Esters by High
Performance Liquid Chromatography (HPLC), Revision 2, 2006.
http://www.epa.gov/epaoswer/hazwaste/test/pdfs/8330b.pdf
5.2.41 EPA ILM05.3 Cyanide: Analytical Methods for Total Cyanide Analysis
This method should be used for preparation and analysis of solid and aqueous liquid samples for the
contaminants identified below and listed in Appendix A. Note: Hydrogen cyanide is not of concern in
solid samples.
Analyte(s)
Cyanide, Total
Hydrogen cyanide
CASRN
57-12-5
74-90-8
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The method allows for either large volume (500-mL aqueous liquid samples or 1-g to 5-g solid samples
mixed with 500 mL of reagent water) or medium volume (50-mL aqueous liquid samples or 1-g solid
samples mixed with 50 mL of reagent water) sample preparation. Aqueous liquid samples are tested for
sulfides and oxidizing agents prior to preparation. Sulfides are removed with cadmium carbonate or lead
carbonate. Samples are treated with sulfuric acid and magnesium chloride and distilled into a sodium
hydroxide solution. The solution is treated with color agents and the cyanide determined as an ion
complex by visible spectrophotometry. The method quantitation limits are 10 (ig/L or 2.5 mg/kg.
Surfactants may interfere with the distillation procedure.
Source: EPA ILM05.3: Exhibit D - Part D: Analytical Methods for Total Cyanide Analysis.
http://www.epa.gov/superfiind/programs/clp/download/ilm/ilm53d.pdf
5.2.42 IO [Inorganic] Compendium Method IO-3.1: Selection, Preparation, and Extraction
of Filter Material
This method should be used for preparation of air samples for the contaminants identified below and
listed in Appendix A. Refer to Appendix A for the appropriate determinative method for these analytes.
Analyte(s)
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
2-Chlorovinylarsonous acid (CVAA)
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]1
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite Oxide
Osmium tetroxide
Sodium arsenite
Thallium sulfate
Vanadium pentoxide
CASRN
7803-55-6
740-38-2
1327-53-3
85090-33-1
541-25-3
40334-69-8
40334-70-1
1306-02-1
20816-12-0
7784-46-5
10031-59-1
1314-62-1
1 Laboratory testing is currently underway for speciation of lewisite 1 using GC-MS techniques.
This method supports determination of arsenic trioxide, lewisite compounds, lewisite oxide, CVAA, and
sodium arsenite as total arsenic. Thallium sulfate is determined as total thallium and ammonium
metavanadate and vanadium pentoxide are determined as total vanadium. A subsample (one-ninth of the
overall filter) is obtained by cutting a strip from the filter used to collect the sample. The filter strip is
extracted using a hydrochloric/nitric acid mix and microwave or hotplate heating. The extract is filtered,
worked up to 20 mL, and analyzed using either Method IO-3.4 or Method IO-3.5.
Source: IO Compendium Method IO-3.1: Compendium of Methods for the Determination of Inorganic
Compounds in Ambient Air: Selection, Preparation and Extraction of Filter Material, 1999.
^
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5.2.43 IO [Inorganic] Compendium Method IO-3.4: Determination of Metals in Ambient
Particulate Matter Using Inductively Coupled Plasma (ICP) Spectroscopy
This method should be used for analysis of air samples for the contaminants identified below and listed in
Appendix A.
Analyte(s)
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
2-Chlorovinylarsonous acid (CVAA)
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]1
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite Oxide
Osmium tetroxide
Sodium arsenite
Thallium sulfate
Vanadium pentoxide
CASRN
7803-55-6
740-38-2
1327-53-3
85090-33-1
541-25-3
40334-69-8
40334-70-1
1306-02-1
20816-12-0
7784-46-5
10031-59-1
1314-62-1
1 Laboratory testing is currently underway for speciation of lewisite 1 using GC-MS techniques.
This method determines arsenic trioxide, lewisite, lewisite degradation products, and sodium arsenite as
total arsenic. Osmium tetroxide is determined as total osmium, thallium sulfate is determined as total
thallium, and ammonium metavanadate and vanadium pentoxide are determined as total vanadium.
Ambient air is sampled by high-volume filters using Method IO-2.1 (a sampling method) and the filters
are extracted by Method IO-3 . 1 . The extracts are analyzed by Inductively Coupled Plasma - Atomic
Emission Spectroscopy (ICP-AES) or Inductively Coupled Plasma - Mass Spectrometry (ICP -MS) (see
Method IO-3.5 in Section 5.2.41). Detection limits, ranges, and interference corrections are dependent on
the analyte and the instrument used.
Source: IO Compendium Method IO-3.4: Compendium of Methods for the Determination of Inorganic
Compounds in Ambient Air: Determination of Metals in Ambient Particulate Matter Using Inductively
Coupled Plasma (ICP) Spectroscopy, 1999.
^
IO Compendium Method IO-2.1: Compendium of Methods for the Determination of Inorganic
Compounds in Ambient Air: Sampling of Ambient Air for Total Suspended Particulate Matter (SPM) and
PM10 Using High Volume (HV) Sampler, 1999.
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5.2.44 IO [Inorganic] Compendium Method IO-3.5: Determination of Metals in Ambient
Particulate Matter Using Inductively Coupled Plasma/Mass Spectrometry (ICP-MS)
This method should be used for analysis of air samples for the contaminants identified below and listed in
Appendix A.
Analyte(s)
Ammonium metavanadate
Arsenic, Total
Arsenic trioxide
2-Chlorovinylarsonous acid (CVAA)
Lewisite 1 (L-1) [2-chlorovinyldichloroarsine]1
Lewisite 2 (L-2) [bis(2-chlorovinyl)chloroarsine]
Lewisite 3 (L-3) [tris(2-chlorovinyl)arsine]
Lewisite Oxide
Sodium arsenite
Thallium sulfate
Vanadium pentoxide
CASRN
7803-55-6
740-38-2
1327-53-3
85090-33-1
541-25-3
40334-69-8
40334-70-1
1306-02-1
7784-46-5
10031-59-1
1314-62-1
1 Laboratory testing is currently underway for speciation of lewisite 1 using GC-MS techniques.
This method determines arsenic trioxide, lewisite, lewisite degradation products, and sodium arsenite as
total arsenic. Thallium sulfate is determined as total thallium and ammonium metavanadate and
vanadium pentoxide are determined as total vanadium. Ambient air is sampled by high-volume filters
using Method IO-2. 1 (a sampling method). The filters are extracted by Method IO-3 . 1 and the extracts
analyzed by Inductively Coupled Plasma/Mass Spectrometry (ICP/MS) or Inductively Coupled Plasma -
Atomic Emission Spectroscopy (ICP-AES) (see Method IO-3.4 in Section 5.2.40). Detection limits,
ranges, and interference corrections are dependent on the analyte and the instrument used.
Source: IO Compendium Method IO-3.5: Compendium of Methods for the Determination of Inorganic
Compounds in Ambient Air: Determination of Metals in Ambient Particulate Matter Using Inductively
Coupled Plasma/Mass Spectrometry (ICP/MS), 1999.
M.g0y7to
IO Compendium Method IO-2.1: Compendium of Methods for the Determination of Inorganic
Compounds in Ambient Air: Sampling of Ambient Air for Total Suspended Particulate Matter (SPM) and
PM10 Using High Volume (HV) Sampler, 1999.
ajjoyTto
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5.2.45 IO [Inorganic] Compendium Method IO-5: Sampling and Analysis for Vapor and
Particle Phase Mercury in Ambient Air Utilizing Cold Vapor Atomic Fluorescence
Spectrometry (CVAFS)
This method should be used for preparation and analysis of air samples for the contaminants identified
below and listed in Appendix A.
Analyte(s)
Mercury, Total
Methoxyethylmercuric acetate
CASRN
7439-97-6
151-38-2
Vapor phase mercury is collected using gold-coated glass bead traps at a flow rate of 0.3 L/min. The
traps are directly desorbed onto a second (analytical) trap. The mercury desorbed from the analytical trap
is determined by Atomic Fluorescence Spectrometry. Particulate mercury is sampled on glass-fiber filters
at a flow rate of 30 L/min. The filters are extracted with nitric acid and microwave heating. The extract
is oxidized with BrCl, then reduced with stannous chloride and purged from solution onto a gold-coated
glass bead trap. This trap is desorbed onto a second trap, the second trap is desorbed, and the mercury is
determined by Atomic Fluorescence Spectrometry. The detection limits are 30 pg/m3 for particulate
mercury and 45 pg/m3 for vapor phase mercury. Detection limits, analytical range, and interferences are
dependent on the instrument used. There are no known positive interferences at 253.7 nm wavelength.
Water vapor will cause a negative interference.
Source: IO Compendium Method IO-5: Compendium of Methods for the Determination of Inorganic
Compounds in Ambient Air: Sampling and Analysis for Vapor and Particle Phase Mercury in Ambient
Air Utilizing Cold Vapor Atomic Fluorescence Spectrometry (CVAFS).
a^^
5.2.46 EPA Air Method, Toxic Organics - 10A (TO-10A): Determination of Pesticides and
Polychlorinated Biphenyls in Ambient Air Using Low Volume Polyurethane Foam
(PUF) Sampling Followed by Gas Chromatographic/Multi-Detector Detection (GC-
MD)
This method should be used for preparation and analysis of air samples for the contaminants identified
below and listed in Appendix A.
Analyte(s)
Chlorfenvinphos
Chlorpyrifos
3-Chloro-1 ,2-propanediol1'2
Chlorosarin1
Chlorosoman1
Cyclohexyl sarin (GF)
Dichlorvos
Dicrotophos
Diisopropyl methylphosphonate (DIMP)1
Dimethylphosphite
Dimethylphosphoramidic acid2
CASRN
470-90-6
2921-88-2
96-24-2
1445-76-7
7040-57-5
329-99-7
62-73-7
141-66-2
1445-75-6
868-85-9
33876-51-6
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Analyte(s)
EA2192 [Diisopropylaminoethyl
methylthiolophosphonate]2
Ethyl methylphosphonic acid (EMPA)2
N-Ethyldiethanolamine (EDEA)
Fenamiphos
Fentanyl
Isopropyl methylphosphonic acid (IMPA)2
Methamidophos3
Methyl parathion
N-Methyldiethanolamine (MDEA)
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Methylphosphonic acid (MPA)2
Mevinphos
Mustard, nitrogen (HN-1) [bis(2-chloroethyl)ethylamine]
Mustard, nitrogen (HN-2) [2,2'-dichloro-N-
methyldiethylamine N,N-bis(2-chloroethyl)methylamine]
Mustard, nitrogen (HN-3) [tris(2-chloroethyl)amine]
Mustard, sulfur/ Mustard gas (HD)
Parathion
Phencyclidine
Phenol
Phorate
Phosphamidon
Pinacolyl methyl phosphonic acid (PMPA)2
R-33 (VR) [methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-methylpropyl ester]
Sarin (GB)1
Semivolatile Organic Compounds, NOS4
Soman (GD)1
Tabun (GA)
Tear gas (CS) [chlorobenzylidene malonitrile]
Tetraethyl pyrophosphate
CASRN
73207-98-4
1832-53-7
139-87-7
22224-92-6
437-38-7
1832-54-8
10265-92-6
298-00-0
105-59-9
1189-87-3
993-13-5
7786-34-7
538-07-8
51-75-2
555-77-1
505-60-2
56-38-2
77-10-1
108-95-2
298-02-2
13171-21-6
616-52-4
159939-87-4
107-44-8
NA
96-64-0
77-81-6
2698-41-1
107-49-3
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Analyte(s)
Tetramethylenedisulfotetramine
Thiodiglycol (TDG)
Triethanolamine (TEA)
Trimethyl phosphite
VE [phosphonothioic acid, ethyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VG [phosphonothioic acid, S-(2-(diethylamino)ethyl)
O,O-diethyl ester]
VM [phosphonothioic acid, methyl-, S-(2-
(diethylamino)ethyl) O-ethyl ester]
VX [O-ethyl-S-(2-diisopropylaminoethyl)methyl-
phosphonothiolate]
CASRN
80-12-6
111-48-8
102-71-6
121-45-9
21738-25-0
78-53-5
21770-86-5
50782-69-9
The following analyte should be determined by this method only if problems (e.g., insufficient recovery,
interferences) occur when using Method TO-15.
Allyl alcohol
107-18-6
1 If problems occur when using this method, it is recommended that the canister Method TO-15 be used.
2 For this analyte, Method TO-10A must be modified to include a derivatization step.
3 If problems occur when using this method, it is recommended that NIOSH Method 5600 be used.
4 NOS = Not otherwise specified
A low-volume (1 to 5 L/minute) sample collection rate is used to collect vapors on a sorbent cartridge
containing polyurethane foam (PUF) or PUF in combination with another solid sorbent. Airborne
particles also are collected, but the sampling efficiency is not known. Pesticides and other chemicals are
extracted from the sorbent cartridge with 5% diethyl ether in hexane and determined by gas
chromatography coupled with an electron capture detector (BCD). A nitrogen-phosphorous detector
(NPD), flame photometric detector (FPD), Hall electrolytic conductivity detector (HECD), or mass
spectrometer (MS) also may be used. Dimethylphosphoramidic acid, EA2192, EMPA, IMPA, MPA, and
PMPA require derivatization with a trimethylsilyl agent prior to injection into the GC. This method is
applicable to multicomponent atmospheres, 0.001 to 50 (ig/m3 concentrations, and 4 to 24-hour sampling
periods. The limit of detection will depend on the specific compounds measured, the concentration level,
and the degree of specificity required. If analytical difficulties are noted, the canister procedures
described in Method TO-15 may be appropriate.
Source: EPA Air Method, Toxic Organics-lOA (TO-10A): Compendium of Methods for the
Determination of Inorganic Compounds in Ambient Air: Determination of Pesticides and Polychlorinated
Biphenyls in Ambient Air Using Low Volume Polyurethane Foam (PUF) Sampling Followed by Gas
Chromatographic/Multi-Detector Detection (GC/MD), 1999.
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5.2.47 EPA Air Method, Toxic Organics -15 (TO-15): Determination of Volatile Organic
Compounds (VOCs) in Air Collected in Specially-Prepared Canisters and Analyzed
by Gas Chromatography/Mass Spectrometry (GC-MS)
This method should be used for preparation and analysis of air samples for the contaminants identified
below and listed in Appendix A.
Analyte(s)
Allyl alcohol1
Carbon disulfide
Cyanogen chloride
1,2-Dichloroethane
Ethyldichloroarsine (ED)
Ethylene oxide
CASRN
107-18-6
75-15-0
506-77-4
107-06-2
598-14-1
75-21-8
The following analytes should be determined by this method only if problems (e.g., insufficient recovery,
interferences) occur when using Method TO-10A.
3-Chloro-1 ,2-propanediol2
Chlorosarin
Chlorosoman
Diisopropyl methylphosphonate (DIMP)
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Sarin (GB)
Soman (GD)
96-24-2
1445-76-7
7040-57-5
1445-75-6
1189-87-3
107-44-8
96-64-0
1 If problems occur when using this method, it is recommended that Method TO-1OA be used.
2 For this analyte, Method TO-15 must be modified to include a derivatization step.
The atmosphere is sampled by introduction of air into a specially prepared stainless steel canister
(specially electropolished or silica-coated). A sample of air is drawn through a sampling train comprising
components that regulate the rate and duration of sampling into the pre-evacuated and passivated canister.
Grab samples also may be collected. After the air sample is collected, the canister valve is closed, an
identification tag is attached to the canister, and the canister is transported to the laboratory for analysis.
To analyze the sample, a known volume of sample is directed from the canister through a solid
multisorbent concentrator. Recovery of less volatile compounds may require heating the canister.
After the concentration and drying steps are completed, VOCs are thermally desorbed, entrained in a
carrier gas stream, and then focused in a small volume by trapping on a cryo-focusing (ultra-low
temperature) trap or small volume multisorbent trap. The sample is then released by thermal desorption
and analyzed by Gas Chromatography/Mass Spectrometry (GC-MS). This method applies to ambient
concentrations of VOCs above 0.5 ppbv and typically requires Volatile Organic Compounds (VOC)
enrichment by concentrating up to 1 L of a sample volume; however, when using current technologies,
quantifications of approximately 100 pptv have been achieved with 0.5-L sample volumes.
Source: EPA Air Method, Toxic Organics-15 (TO-15): Compendium of Methods for the Determination
of Inorganic Compounds in Ambient Air: Determination of Volatile Organic Compounds (VOCs) in Air
Collected in Specially-Prepared Canisters and Analyzed by Gas Chromatography/Mass Spectrometry
(GC/MS), 1999.
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5.2.48 NIOSH Method 1612: Propylene Oxide
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Propylene oxide
CASRN
75-56-9
A sample tube containing coconut shell charcoal is used for sample collection with a flow rate of 0.01 to
0.2 L/min. One milliliter of carbon disulfide (CS2) is added to the vial and allowed to sit for 30 minutes
prior to analysis with occasional agitation. No interferences have been found. The working range is
between 8 to 295 ppm for air samples of 5 L.
Source: NIOSH Method 1612: Propylene Oxide, Issue 2, 1994.
http: //www. cdc. go v/niosh/nmam/pdfs/1612 .pdf
5.2.49 NIOSH Method 2016: Formaldehyde
This method should be used for preparation and analysis of samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Formaldehyde
CASRN
50-00-0
This method can be used for the determination of formaldehyde using high performance liquid
chromatography (HPLC) with an ultraviolet (UV) detector. Air is sampled onto a cartridge containing
silica gel coated with 2,4-dinitrophenyl hydrazine, at a rate of 0.03 to 1.5 L/min. The cartridge is
extracted with 10 mL of acetonitrile and analyzed by HPLC-UV at a wavelength of 360 nm. The working
range is 0.015 to 2.5 mg/m3 (0.012 to 2.0 ppm) for a 15-L sample. The detection limit for formaldehyde
is 0.07 (ig/sample. Ozone has been observed to consume the 2,4-dinitrophenylhydrazine (2,4-DNPH)
reagent and to degrade the formaldehyde derivative. Ketones and other aldehydes can react with 2,4-
DNPH; the derivatives produced, however, are separated chromatographically from the formaldehyde
derivative.
Source: NIOSH Method 2016: Formaldehyde, Issue 2, 2003.
5.2.50 NIOSH Method 2513: Ethylene Chlorohydrin
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
2-Chloroethanol (ethylene chlorohydrin)
CASRN
107-07-3
Samples are drawn into a tube containing petroleum charcoal at a rate of 0.01 to 0.2 L/min and transferred
into vials containing eluent (carbon disulfide, 2-propanol, and «-pentadiene as an internal standard).
Vials must sit for 30 minutes prior to analysis by gas chromatography - flame ionization detection (GC-
FID). No interferences have been identified. Humidity may decrease the breakthrough volume during
sample collection. The working range of the method is 0.5 to 15 ppm for a 20-L air sample.
SAM Revision 3.1 71 November 15, 2007
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Section 5 - Selected Chemical Methods
Source: NIOSH Method 2513: Ethylene Chlorohydrin, Issue 2, 1994.
http://www.cdc.gov/niosh/nmaiii/pdfs/2513 .pdf
5.2.51 NIOSH Method 3510: Monomethylhydrazine
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Methyl hydrazine (monomethylhydrazine)
CASRN
60-34-4
Samples are collected into a bubbler containing HC1 using a flow rate of 0.5 to 1.5 L/min. Samples are
then mixed with phosphomolybdic acid solution and transferred to a large test tube for spectrophotometric
analysis. Positive interferences that have been noted include stannous ion, ferrous ion, zinc, sulfur
dioxide, and hydrogen sulfide. Negative interferences may occur by oxidation of the
monomethylhydrazine by halogens, oxygen (especially in the presence of copper (I) ions) and hydrogen
dioxide. The working range of the method is 0.027 to 2.7 ppm for a 20-L air sample.
Source: NIOSH Method 3510: Monomethylhydrazine, Issue 1, 1994.
http: //www. cdc. go v/niosh/nmam/pdfs/3 510 .pdf
5.2.52 NIOSH Method 5600: Organophosphorus Pesticides
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Organophosphate Pesticides, NOS1
CASRN
NA
The following analyte should be determined by this method only if problems (e.g., insufficient recovery,
interferences) occur when using Method TO-10A.
Methamidophos
10265-92-6
NOS = Not otherwise specified
This method is used for the detection of Organophosphorus pesticides using gas chromatography (GC)
with a flame photometric detector (FPD). The method also may be applicable to the determination of
other Organophosphorus compounds after evaluation for desorption efficiency, sample capacity, sample
stability, and precision and accuracy. The working range for each analyte is provided in Table 5 of the
method. These ranges cover from 0.1 to 2 times the OSHA Permissible Exposure Limits (PELs) (see
Table 5 of the method). The method also is applicable to Short Term Exposure Limit (STEL)
measurements using 12-L samples. The detection limit depends on the compound being analyzed.
Several organophosphates may co-elute with either target analytes or internal standards causing
integration errors. These include other pesticides, and the following: tributyl phosphate, tris-(2-butoxy
ethyl) phosphate, tricresyl phosphate, and triphenyl phosphate.
Source: NIOSH Method 5600: Organophosphorus Pesticides, Issue 1, 1994.
http://www.cdc.gov/niosh/nmam/pdfs/5600.pdf
SAM Revision 3.1 72 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.53 NIOSH Method 5601: Organonitrogen Pesticides
This method should be used for preparation and analysis of air samples for the contaminants identified
below and listed in Appendix A.
Analyte(s)
Aldicarb
Carbofuran
Methomyl
Oxamyl
CASRN
116-06-3
1563-66-2
16752-77-5
23135-22-0
This method can be used for the determination of organonitrogen pesticides using high performance
liquid chromatography (HPLC) with an ultraviolet (UV) detector. The method also may be applicable to
the determination of other organonitrogen compounds and to a broad range of pesticides having UV
chromophores, e.g., acetanilides, acid herbicides, organophosphates, phenols, pyrethroids, sulfonyl ureas,
sulfonamides, triazines, and uracil pesticides. The detection limit for aldicarb is 1.2 (ig per sample and
0.6 (ig per sample for carbofuran, methyomyl, and oxamyl. The working ranges for aldicarb, carbofuran,
and oxamyl range from 0.5 to 10 times the OSHA Permissible Exposure Limit (PEL) (see Table 2 of the
method). Because of the broad response of the UV detector at shorter wavelengths, there are many
potential interferences. Those tested include solvents (chloroform and toluene), antioxidants (BHT),
plasticizers (dialkyl phthalates), nitrogen compounds (nicotine and caffeine), impurities in HPLC reagents
(e.g., in triethylamine), other pesticides (2,4-D, atrazine, parathion, etc.), and pesticide hydrolysis
products (1-naphthol). Confirmation techniques are recommended when analyte identity is uncertain.
Source: NIOSH Method 5601: Organonitrogen Pesticides, Issue 1, 1998.
icj>OT/nioj^
5.2.54 NIOSH Method 6001: Arsine
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Arsine
CASRN
7784-42-1
Arsine is determined as arsenic. A 0.1 to 10-L volume of air is drawn through a sorbent tube containing
activated charcoal. The sorbent is extracted with a nitric acid solution, and arsenic is determined by
graphite furnace atomic absorption. The working range of the method is 0.001 to 0.2 mg/m3 for a 10-L
sample. The method is subject to interferences from other arsenic compounds.
Source: NIOSH Method 6001: Arsine, Issue 2, 1994. http://www.cdc.gov/niosli/nmaiii/pdfs/6001 .pdf
SAM Revision 3.1 73 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.55 NIOSH Method 6002: Phosphine
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Phosphine
CASRN
7803-51-2
In this method, phosphine is determined as phosphate. A volume of 1 to 16 L of air are drawn through a
sorbent tube containing silica gel coated with Hg(CN)2. The sorbent is extracted with a potassium
permanganate/ sulfuric acid solution and washed with reagent water. Following treatment with the color
agent and extraction into organic solvent, phosphate is determined by visible spectrometry. The working
range of the method is 0.02 to 0.9 mg/m3 for a 16-L sample. The method is subject to interferences from
phosphorus trichloride, phosphorus pentachloride, and organic phosphorus compounds.
Source: NIOSH Method 6002: Phosphine, Issue 2, 1994. http://www.cdc.gov/niosli/nmam/pdfs/6002.pdf
5.2.56 NIOSH Method 6004: Sulfur Dioxide
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Sulfur dioxide
CASRN
7446-09-5
In this method, sulfur dioxide is determined as sulfite plus sulfate. A volume of 40 to 200 L of air is
drawn through a sodium carbonate-treated filter that is preceded by a 0.8 (im filter to remove particulates
and sulfuric acid. The treated filter is extracted with a carbonate/bicarbonate solution and the extract
analyzed by ion chromatography for sulfite and sulfate. The sulfur dioxide is present as sulfite on the
filter; however, because sulfite oxidizes to sulfate, both ions must be determined and the results summed.
The working range of the method is 0.5 to 20 mg/m3 for a 100-L sample. The method is subject to
interference from sulfur trioxide in dry conditions.
Source: NIOSH Method 6004: Sulfur Dioxide, Issue 2, 1994.
5.2.57 NIOSH Method 6010: Hydrogen Cyanide
This method should be used for preparation and analysis of air samples for the contaminants identified
below and listed in Appendix A.
Analyte(s)
Cyanide, Total
Hydrogen cyanide
CASRN
57-12-5
74-90-8
Hydrogen cyanide is determined as a cyanide ion complex by this method. A volume of 2 to 90 L of air
is drawn through a soda lime sorbent tube. A glass-fiber filter is used to remove particulate cyanides
prior to the sorbent tube. Cyanide is extracted from the sorbent with reagent water treated with NaOH.
The extract is pH adjusted with HC1, oxidized with N-chlorosuccinimide/succinimide, and treated with
the coupling-color agent (barbituric acid/pyridine). The cyanide ion is determined by visible
spectrophotometry. The working range of the method is 3 to 260 mg/m3 for a 3-L sample. The method is
subject to interference from high concentrations of hydrogen sulfide.
SAM Revision 3.1 74 November 15, 2007
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Section 5 - Selected Chemical Methods
Source: NIOSH Method 6010: Hydrogen Cyanide, Issue 2, 1994.
http: //www. cdc. go v/niosh/nmam/pdfs/6010 .pdf
5.2.58 NIOSH Method 6013: Hydrogen Sulfide
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Hydrogen sulfide
CASRN
7783-06-4
Hydrogen sulfide is determined as sulfate by this method. A volume of 1.2 to 40 L of air is drawn
through charcoal sorbent. A prefilter is used to remove particulates. The sorbent portions are extracted
with an ammonium hydroxide/hydrogen peroxide solution and the extract is analyzed for sulfate by ion
chromatography. The working range of the method is 0.9 to 20 mg/m3 for a 20-L sample. The method is
subject to interference from sulfur dioxide.
Source: NIOSH Method 6013: Hydrogen Sulfide, Issue 1, 1994.
http://www.cdc.gov/niosh/nmaiii/pdfs/6013 .pdf
5.2.59 NIOSH Method 6015: Ammonia
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Ammonia
CASRN
7664-41-7
Ammonia is determined as indophenol blue by this method. A volume of 0.1 to 96 L of air is drawn
through a sulfuric acid-treated silca gel sorbent. A prefilter is used to remove particulates. The sorbent is
extracted with reagent water, the pH adjusted, and reagents are added to generate the indophenol blue
compound in the presence of ammonium. The extract is analyzed by visible spectrophotometry. The
working range of the method is 0.15 to 300 mg/m3 for a 10-L sample. Twice the recommended sample
volume should be collected in order to achieve an action level of 70 (ig/m3. No interferences have been
identified.
Source: NIOSH Method 6015: Ammonia, Issue 2, 1994. http://www.cdc.gov/niosh/nmam/pdfs/6015.pdf
5.2.60 NIOSH Method 6402: Phosphorus Trichloride
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Phosphorus trichloride
CASRN
7719-12-2
In this method, phosphorus trichloride is determined as phosphate. A volume of 11 to 100 L of air is
drawn through a bubbler containing reagent water. The resulting H3PO3 solution is oxidized to H3PO4
and color agents are added. The solution is analyzed by visible spectrophotometry. The working range of
the method is 1.2 to 80 mg/m3 for a 25-L sample. Phosphorus (V) compounds do not interfere. The
sample solutions are stable to oxidation by air during sampling.
SAM Revision 3.1 75 November 15, 2007
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Section 5 - Selected Chemical Methods
Source: NIOSH Method 6402: Phosphorus Trichloride, Issue 2, 1994.
http: //www. cdc. go v/niosli/nmaiii/pdfs/6402 .pdf
5.2.61 NIOSH Method 7903: Acids, Inorganic
This method should be used for preparation and analysis of air samples for the contaminants identified
below and listed in Appendix A.
Analyte(s)
Hydrogen bromide
Hydrogen chloride
Hydrogen fluoride1
CASRN
10035-10-6
7647-01-0
7664-39-3
1 If problems occur when using this method, it is recommended that NIOSH 7906 be used.
Acids are analyzed as bromide, chloride, and fluoride, respectively, by this method. A volume of 3 to 100
L of air is drawn through a silica gel sorbent. The sorbent portions are extracted with a buffered
carbonate/bicarbonate solution and the extract is analyzed by ion chromatography. The working range of
this method is 0.01 to 5 mg/m3 for a 50-L sample. Particulate salts of the acids are an interference
(trapped on the glass wool filter plug in the sorbent tube). Chlorine and bromine are also interferences.
Acetate, formate, and propionate interferences may be reduced by use of a weaker eluent.
Source: NIOSH Method 7903: Acids, Inorganic, Issue 2, 1994.
http://www.cdc.gov/niosh/nmam/pdfs/7903.pdf
5.2.62 NIOSH Method 7905: Phosphorus
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
White phosphorus
CASRN
12185-10-3
This method identifies and determines the concentration of white phosphorus in air by using a gas
chromatography/flame photometric detector (GC-FPD). Samples prepared by this method can be
analyzed alternatively by gas chromatography/nitrogen phosphorus detection (GC-NPD). Five to 100 L
of air are drawn through a GC solid sorbent tube, and the sorbent is extracted (desorbed) with xylene.
Phosphorus is determined by GC-FPD or GC-NPD. The working range for samples analyzed by GC-
FPD is 0.056 to 0.24 mg/m3 for a 12-L sample. The limit of detection (LOD) for samples analyzed by
GC-FPD is 0.005 ug per sample. The method is applicable to vapor-phase phosphorus only; if particulate
phosphorus is expected, a filter could be used in the sampling train.
Source: NIOSH Method 7905: Phosphorus, Issue 2, 1994.
SAM Revision 3.1 76 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.63 NIOSH Method 7906: Fluorides, Aerosol and Gas
This method is not currently listed for any of the analyte/sample type combinations included in Appendix
A. If problems occur when using NIOSH Method 7903 for the analysis of hydrogen fluoride, then this
method should be used for preparation and analysis of air samples for the contaminant identified below
and listed in Appendix A. (See Footnote 9 of Appendix A.)
Analyte(s)
Hydrogen fluoride1
CASRN
7664-39-3
1 This analyte should be determined by this method (7906) only if problems occur when using NIOSH Method 7903.
Hydrogen fluoride is determined as fluoride ion by this method. A volume of 1 to 800 L of air is drawn
through a 0.8-(im cellulose ester membrane (to trap particulate fluorides) and a cellulose pad treated with
sodium carbonate (to trap gaseous fluoride). The pad is extracted with reagent water and the extract is
analyzed for fluoride by ion chromatography. The working range of the method is 0.04 to 8 mg/m3 for
250-L samples. If other aerosols are present, gaseous fluoride may be slightly underestimated due to
adsorption onto or reaction with particles, with concurrent overestimation of particulate/gaseous fluoride
ratio.
Source: NIOSH Method 7906: Fluorides, Aerosol and Gas by 1C, Issue 1, 1994.
http: //www. cdc. go v/niosh/nmam/pdfs/7906 .pdf
5.2.64 NIOSH Method S301-1: Fluoroacetate Anion
This method should be used for preparation of air samples for the contaminant identified below and
listed in Appendix A. Note: EPA Method 300.1 Rev 1.0 should be used as the determinative method
(refer to Appendix A).
Analyte(s)
Fluoroacetic acid and fluoroacetate salts
Methyl fluoroacetate
CASRN
NA
453-18-9
This method was developed specifically for sodium fluoroacetate, but also may be applicable to other
fluoroacetate salts. The method determines fluoroacetate salts as fluoroacetate anion. A known volume
of air (e.g., 480 L was used in validation of this method) is drawn through a cellulose ester membrane
filter to collect sodium fluoroacetate. Sodium fluoroacetate is extracted from the filter with 5 mL of
deionized water, and the resulting sample is analyzed by ion chromatography using electrolytic
conductivity detection. The analytical range of this method is estimated to be 0.01 to 0.16 mg/m3. The
detection limit is estimated to be 20 ng of sodium fluoroacetate per injection, corresponding to a 100-uL
aliquot of a 0.2-ug/mL standard.
Source: NIOSH Method S301-1: Sodium Fluoroacetate, 1977. httg7/www.cdc.goy/niosh/gdfe/s3_OLgdf
5.2.65 OSHA Method 40: Methylamine
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Methylamine
CASRN
74-89-5
This method is used for detection of methylamine using high performance liquid chromatography (HPLC)
with a fluorescence (FL) or visible (vis) detector. Samples are collected by drawing 10-L volumes of air
SAM Revision 3.1 77 November 15, 2007
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Section 5 - Selected Chemical Methods
at a rate of 0.2 L/min through standard size sampling tubes containing XAD-7 resin coated with 10%
NBD chloride by weight. Samples are desorbed with 5% (w/v) 7-chloro-4-nitrobenzo-2-oxa-l,3-diazole
(NBD chloride) in tetrahydrofuran (with a small amount of sodium bicarbonate present), heated in a hot
water bath, and analyzed by HPLC-FL or HPLC-vis. The detection limit of the overall procedure is 0.35
ug per sample (28 ppb or 35 ug/m3). Quantitation limits of 28 ppb (35 ug/m3) have been reliably
achieved. This is the smallest amount of methylamine that can be quantified within the requirements of a
recovery of at least 75% and a precision (standard deviation of 1.96) of ±25% or belter.
Source: OSHA Method 40: Methylamine, 1982.
Iittp://www.osha.gov/dts/sltc/mctliods/organic/org040/org040.html
5.2.66 OSHA Method 54: Methyl Isocyanate
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Methyl isocyanate
CASRN
624-83-9
This method determines the concentration of methyl isocyanate in air by using high performance liquid
chromatography (HPLC) with a fluorescence or ultraviolet (UV) detector. Samples are collected by
drawing a known volume of air through XAD-7 tubes coated with 0.3 mg of l-(2-pyridyl)piperazine (1-
2PP). Samples are desorbed with acetonitrile (ACN) and analyzed by HPLC using a fluorescence or UV
detector.
Source: OSHA Method 54: Methyl Isocyanate (MIC), 1985.
http://www.osha.gov/dts/sltc/methods/organic/org054/org054.html
5.2.67 OSHA Method 61: Phosgene
This method should be used for preparation and analysis of air samples for the contaminants identified
below and listed in Appendix A.
Analyte(s)
Perfluoroisobutylene (PFIB)1
Phosgene
CASRN
382-21-8
75-44-5
If problems occur when using this method, it is recommended that a method based on the following journal article
be used: J. Chrom. A, 1098: (2005) 156-165.
This method determines the concentration of phosgene in air by using gas chromatography with a
nitrogen selective detector. Air samples are collected by drawing known volumes of air through sampling
tubes containing XAD-2 adsorbent that has been coated with 2-(hydroxymethyl)piperidine. The samples
are desorbed with toluene and then analyzed by gas chromatography using a nitrogen selective detector.
Source: OSHA Method 61: Phosgene, 1986.
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SAM Revision 3.1 78 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.68 OSHA Method ID-211: Sodium Azide and Hydrazoic Acid in Workplace
Atmospheres
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Sodium azide
CASRN
26628-22-8
This method describes sample collection and analysis of airborne azides [as sodium azide (NaN3) and
hydrazoic acid (HN3)]. Particulate sodium azide (NaN3) is collected on a PVC filter or in the glass wool
plug of the sampling tube. Gaseous HN3 is collected and converted to NaN3 by the impregnated silica gel
(ISO) sorbent within the sampling tube. The collected azide on either media is desorbed in a weak buffer
solution, and the resultant anion (N3~) is analyzed by Ion Chromatography (1C) using a variable
wavelength UV detector at 210 nm. A gravimetric conversion is used to calculate the amount of NaN3 or
HN3 collected. The detection limit was found to be 0.001 ppm as HN3 or 0.003 mg/m3 as NaN3 for a 5-L
air sample. The quantitation limit was found to be 0.004 ppm as HN3 or 0.011 mg/m3 as NaN3 for a 5-L
air sample.
Source: OSHA Method ID-211: Sodium Azide and Hydrazoic Acid in Workplace Atmospheres.
a^^
5.2.69 OSHA Method ID-216SG: Boron Trifluoride (BF3)
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Boron trifluoride
CASRN
7637-07-2
Boron trifluoride is determined as fluoroborate by this method. A volume of 30 to 480 L of air is drawn
through a bubbler containing 0.1 M ammonium fluoride. The solution is diluted and analyzed with a
fluoroborate ion specific electrode (ISE). The detection limit is 10 (ig in a 30-L sample.
Source: OSHA Method ID-216SG: Boron Trifluoride (BF3), 1989.
http://www.osha.gov/dts/sltc/methods/partial/id216sg/id216sg.html
5.2.70 OSHA Method PV2004: Acrylamide
This method should be used for preparation and analysis of air samples for the contaminants identified
below and listed in Appendix A.
Analyte(s)
Acrylamide
Acrylonitrile
Methylacrylonitrile
CASRN
79-06-1
107-13-1
126-98-7
This method determines the concentration of acrylamide in air by using high performance liquid
chromatography (HPLC) with an ultraviolet (UV) detector. Samples are collected by drawing known
SAM Revision 3.1 79 November 15, 2007
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Section 5 - Selected Chemical Methods
volumes of air through OSHA versatile sampler (OVS-7) tubes, each containing a glass fiber filter and
two sections of XAD-7 adsorbent. Samples are desorbed with a solution of 5% methanol/95% water and
analyzed by HPLC using a UV detector. The detection limit was found to be 0.7 (ig/mL (0.006 mg/m3 for
a 1-mL desorption volume or 0.029 mg/m3 for a 5-mL desorption volume based on a 120-L air volume).
Applicable working ranges for a 1-mL and 5-mL desorption volume are 0.017 - 1.5 mg/m3 and 0.083 -
7.5 mg/m3, respectively.
Source: OSHA Method PV2004: Acrylamide.
5.2.71 OSHA Method PV2103: Chloropicrin
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Chloropicrin
CASRN
79-06-2
This method determines the concentration of Chloropicrin in air by gas chromatography/electron capture
detection (GC-ECD). Samples are collected by drawing a known volume of air through two XAD-4
tubes in series. Samples are desorbed with ethyl acetate and analyzed by GC-ECD. Chloropicrin is light
sensitive, and samples should be protected from light. The detection limit of the analytical procedure is
0.01 ng, with a l-(iL injection volume. This is the smallest amount that could be detected under normal
operating conditions. The working range is 0.25 to 0.99 ppmv.
Source: OSHA Method PV2103: Chloropicrin.
01-9111 -ch/t-pv2103-01-9111 -ch.html
5.2.72 ASTM Method D5755-03: Standard Test Method for Microvacuum Sampling and
Indirect Analysis of Dust by Transmission Electron Microscopy (TEM) for
Asbestos Structure Number Surface Loading
This method should be used for preparation and analysis of solid samples (e.g., soft surfaces-microvac)
for the contaminant identified below and listed in Appendix A.
Analyte(s)
Asbestos
CASRN
1332-21-4
This method describes procedures to identify asbestos in dust and provide an estimate of the surface
loading of asbestos reported as the number of asbestos structures per unit area of sampled surface. The
sample is collected by vacuuming a known surface area with a standard 25- or 37-mm air sampling
cassette using a plastic tube that is attached to the inlet orifice, which acts as a nozzle. The sample is
transferred from inside the cassette to an aqueous suspension of known volume. Aliquots of the
suspension are then filtered through a membrane, and a section of the membrane is prepared and
transferred to a transmission electron microscopy (TEM) grid using a direct transfer method. The
asbestiform structures are identified, sized, and counted by TEM, using select area electron diffraction
(SAED) and energy dispersive X-ray analysis (EDXA) at a magnification of 15,000 to 20,OOOX.
SAM Revision 3.1 80 November 15, 2007
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Section 5 - Selected Chemical Methods
Source: ASTM Method D5755-03: Standard Test Method for Microvacuum Sampling and Indirect
Analysis of Dust by Transmission Electron Microscopy for Asbestos Structure Number Surface Loading,
2003. http://www.astm.org/c.gi-bin/SoftCart.cxc/STORE/filtrcxx40.c.gi?U+mystorc+tavs3076+-
L+D5755:03+/usr6/htdocs/astm.org/DATABASE.CART/REDLINE_PAGES/D5755.htm
5.2.73 ASTM Method D6480-99: Standard Test Method for Wipe Sampling of Surfaces,
Indirect Preparation, and Analysis for Asbestos Structure Number Concentration
by Transmission Electron Microscopy
This method should be used for preparation and analysis of solid samples (e.g., hard surfaces-wipes) for
the contaminant identified below and listed in Appendix A.
Analyte(s)
Asbestos
CASRN
1332-21-4
This method describes a procedure to identify asbestos in samples wiped from surfaces and to provide an
estimate of the concentration of asbestos reported as the number of asbestos structures per unit area of
sampled surface. A sample is collected by wiping a surface of known area with a wipe material. The
sample is transferred from the wipe material to an aqueous suspension of known volume. Aliquots of the
suspension are then filtered through a membrane filter, and a section of the membrane filter is prepared
and transferred to a TEM grid, using the direct transfer method. The asbestiform structures are identified,
sized, and counted by TEM, using electron diffraction (ED) and energy dispersive X-ray analysis
(EDXA) at a magnification from 15,000 to 20,OOOX.
Source: ASTM Method D6480-99: Standard Test Method for Wipe Sampling of Surfaces, Indirect
Preparation, and Analysis for Asbestos Structure Number Concentration by Transmission Electron
Microscopy. http://ww\\'.astm.org/cgi-bin/SoftCart.cxc/STORE/filtrcxx40.cgi?U+mystorc+tavs3076+-
L+D6480:99+/usr6/htdocs/astm.om/DATABASE.CART/HISTORICAL/D6480-99.htm
5.2.74 ISO Method 10312:1995: Ambient Air- Determination of Asbestos Fibres - Direct-
transfer Transmission Electron Microscopy Method (TEM)
This method should be used for preparation and analysis of air samples for the contaminant identified
below and listed in Appendix A.
Analyte(s)
Asbestos
CASRN
1332-21-4
This method determines the type(s) of asbestos fibers present, but cannot discriminate between individual
fibers of the asbestos and non-asbestos analogues of the same amphibole mineral. The method is defined
for polycarbonate capillan/pore filters or cellulose ester (either mixed esters of cellulose or cellulose
nitrate) filters through which a known volume of air has been drawn. The method is suitable for
determination of asbestos in both exterior and building atmospheres. The range of concentrations that can
be determined is 50 structures/mm2 to 7,000 structures/mm2 on the filter. In a 4000-L air sample with
approximately 10 pg/m3 (typical of clean or rural atmospheres), an analytical sensitivity of 0.5 structure/L
can be obtained. This is equivalent to a detection limit of 1.8 structure/L when an area of 0.195 mm of
the TEM specimen is examined.
Source: ISO Method 10312: 1995: Ambient Air—Determination of Asbestos Fibres—Direct Transfer
Transmission Electron Microscopy Method, 2005.
httK//wwwiso^^
40&ICS3=20
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Section 5 - Selected Chemical Methods
5.2.75 Standard Method 4500-NH3 B: Nitrogen (Ammonia) Preliminary Distillation Step
This method should be used for preparation of aqueous/liquid samples for the contaminant identified
below and listed in Appendix A. Note: Standard Method 4500-NH3 G should be used as the
determinative method (refer to Appendix A).
Analyte(s)
Ammonia
CASRN
7664-41-7
A 0.5- to 1-L sample is dechlorinated, buffered, adjusted to pH 9.5, and distilled into a sulfuric acid
solution. The distillate is brought up to volume, neutralized with sodium hydroxide, and analyzed by
Method 4500-NH3 G.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition, http://www.standardmcthods.org/
5.2.76 Standard Method 4500-NH3 G: Nitrogen (Ammonia) Automated Phenate Method
This method should be used for analysis of aqueous liquid samples for the contaminant identified below
and listed in Appendix A.
Analyte(s)
Ammonia
CASRN
7664-41-7
Ammonia is determined as indophenol blue by this method. A portion of the neutralized sample distillate
(from procedure 4500-NH3 B) is run through a manifold. The ammonium in the distillate reacts with
solutions of disodium ethylenediaminetetraacetic acid (EDTA), sodium phenate, sodium hypochlorite,
and sodium nitroprusside. The resulting indophenol blue is detected by colorimetry in a flow cell. The
range of the method is 0.02 to 2.0 mg/L.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition, http://www.standardmcthods.org/
5.2.77 Standard Method 4500-CI G: DPD Colorimetric Method
This method should be used for preparation and analysis of aqueous liquid and drinking water samples
for the contaminant identified below and listed in Appendix A. It also should be used for analysis of air
samples when appropriate sample preparation techniques have been applied (refer to Appendix A).
Analyte(s)
Chlorine
CASRN
7782-50-5
A portion of aqueous liquid sample is buffered and reacted with N,N-diethyl-/>-phenylenediamine (DPD)
color agent. The resulting free chlorine is determined by colorimetry. If total chlorine (including
chloroamines and nitrogen trichloride) is to be determined, KI crystals are added. Results for chromate
and manganese are blank corrected using thioacetamide solution. The method can detect 10 (ig/L
chlorine. Organic contaminants and strong oxidizers may cause interference.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition. littBi//wjvwjjtea
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Section 5 - Selected Chemical Methods
5.2.78 Literature Reference for Chlorine (Analyst, 1999. 124: 1853-1857)
This procedure should be used for preparation of air samples for the contaminant identified below and
listed in Appendix A. Note: Standard Method 4500-C1 G should be used as the determinative method
(refer to Appendix A).
Analyte(s)
Chlorine
CASRN
7782-50-5
A procedure is described for determination of total combined gas-phase active chlorine (i.e., C12, HOC1
and chloramines) and is based on a sulfonamide-functionalized silica gel sorbent. For determination of
the collected chlorine, a modified version of the N,N-diethyl-/>-phenylenediamine (DPD) colorimetric
procedure is used, which yielded a detection limit of 0.1 mg of chlorine. At flow rates ranging from 31 to
294 mL/min, the collection efficiency was >90% based on breakthrough analysis. Recovery of chlorine
spikes from 0.05-g aliquots of the sorbent was not quantitative (-60%) but was reproducible; the recovery
is accounted for in samples by adding weighed amounts of sorbent to the standards.
Source: Johnson, B.J., Emerson, D.W., Song, L., Floyd, J., and Tadepalli, B. "Determination of active
chlorine in air by bonded phase sorbent collection and spectophotometric analysis," Analyst, 124: 1853—
1857(1999).
5.2.79 Literature Reference for Fluoroacetate salts (Analytical Letters, 1994. 27 (14):
2703-2718)
The initial portion of this procedure (ultrasonic extraction) should be used for preparation of solid and
non-aqueous liquid/organic solid samples for the contaminant identified below and listed in Appendix A.
Note: EPA Method 300.1, Revision 1.0 should be used as the determinative method (refer to Appendix
A).
Analyte(s)
Fluoroacetic acid and fluoroacetate salts
Methyl fluoroacetate
CASRN
NA
453-18-9
Sodium fluoroacetate is determined at sub-microgram per gram concentrations in small (~1 g) soil
samples. Samples are ultrasonically extracted with water, filtered, and analyzed by Method 300.1.
Source: Tomkins, B.A., "Screening-Procedure for Sodium Fluoroacetate (Compound 1080) at Sub-
Microgram/Gram Concentrations in Soils," Analytical Letters. 27(14), 2703—2718 (1994).
SAM Revision 3.1 83 November 15, 2007
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Section 5 - Selected Chemical Methods
5.2.80 Literature Reference for Perfluoroisobutylene (Journal of Chromatography A,
2005. 1098: 156-165)
This procedure is not currently listed for any of the analyte/sample type combinations included in
Appendix A. If problems occur when using OSHA Method 61 for the analysis of perfluoroisobutylene,
then this procedure should be used for preparation and analysis of air samples for the contaminant
identified below and listed in Appendix A. (See Footnote 15 of Appendix A.)
Analyte(s)
Perfluoroisobutylene (PFIB)1
CASRN
382-21-8
This analyte should be determined using this article only if problems occur when using OSHA Method 61.
This procedure is for the identification and measurement of phosgene and perfluoroisobutylene in air
using gas chromatography/mass spectrometry (GC-MS), gas chromatography/nitrogen phosphorus
detector (GC-NPD), or gas chromatography/flame photometric detector (GC-FPD). This procedure is to
be used in the event that OSHA Method 61 is problematic for the determination of PFIB. Air samples are
collected by drawing known volumes of air through sampling tubes containing Tenax TA sorbent coated
with 2-aminothiophenol (ATP) or 3,4-dimercaptotoluene (DMT). The derivatized samples are thermally
desorbed and analyzed with one of the GC techniques. The 2-aminothiophenol derivative can be
analyzed by either of the three techniques, whereas the 3,4-dimercaptotoluene is not suitable for analysis
by GC-NPD or GC-FPD. Limits of detection for PFIB-ATP, and PFIB-DMT, using 10-L air samples
(typical sampling volume) by GC-MS analyses were determined to be 2 and 19 ng/m3 respectively.
Source: Muir, B., Cooper, D.B., Carrie, W.A., Timperley, C.M., Slater, B.J., and S. Quick, "Analysis of
Chemical Warfare Agents III. Use of bis-Nucleophiles in the Trace Level Determination of Phosgene and
Perfluoroisobutylene" Journal of Chromatography A.. 1098, 156-165 (2005).
5.2.81 Literature Reference for Sodium Azide (Journal of Forensic Science, 1998.
43(1):200-202)
This procedure should be used for preparation of solid, non-aqueous liquid/organic solid, aqueous
liquid, and drinking water samples for the contaminant identified below and listed in Appendix A.
Analyte(s)
Sodium azide
CASRN
26628-22-8
Sample aliquots are acidified, converting azide to volatile hydrazoic acid (HN3), which is then trapped in
sodium hydroxide. The resulting aliquots are analyzed by ion chromatography, using a sodium
tetraborate eluent and suppressed conductivity detection. This method can routinely quantify to at least
100 (ig/L, and the detection limit is estimated to be 30 (ig/L. Water extraction, filtration, and acidification
steps should be used for the preparation of solid samples. Filtration and acidification steps should be used
for preparation of aqueous liquid and drinking water samples. The acidification step should be used with
EPA SW-846 Method 3580A for preparation of non-aqueous liquid/organic solid samples.
Please note: The procedure described above has been developed for the analysis of sodium azide in
blood samples.
Source: Kruszyna R, Smith RP, Kruszyna H. "Determining Sodium Azide Concentration in the Blood by
Ion Chromatography," Journal of Forensic Science. 43(1):200-202 (1998).
SAM Revision 3.1 84 November 15, 2007
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Section 6 - Selected Radiochemical Methods
Section 6.0: Selected Radiochemical Methods
A list of analytical methods to be used in analyzing environmental samples for radiochemical
contaminants during homeland security events is provided in Appendix B. Methods are listed for each
isotope and for each sample type that potentially may need to be measured and analyzed when responding
to an environmental emergency.
Please note: This section provides guidance for selecting radiochemical methods that have a high
likelihood of assuring analytical consistency when laboratories are faced with a large scale
environmental restoration crisis. Not all methods have been verified for the analyte/sample type
combination listed in Appendix B. Please refer to the specified method to identify analyte/sample type
combinations that have been verified. Any questions regarding information discussed in this section
should be addressed to the appropriate contact(s) listed in Section 4.
Appendix B is sorted alphabetically by analyte and includes the following information:
• Analyte(s). The radionuclide(s) or contaminant(s) of interest.
• Chemical Abstract Survey Registration Number (CAS RN). A unique identifier for chemical
substances that provides an unambiguous way to identify a chemical or molecular structure when
there are many possible systematic, generic, or trivial names. In this section (Section 6.0) and
Appendix B, the CAS RNs correspond to the specific radionuclide identified.
• Determinative technique. An analytical instrument or technique used for qualitative and
confirmatory determination of compounds or components in a sample.
Drinking water sample methods. The recommended methods/procedures for sample preparation
and analysis to measure the analyte of interest in drinking water samples. Methods have been
identified for qualitative and confirmatory determination.
• Aqueous and liquid phase sample methods. The recommended methods/procedures for sample
preparation and analysis to measure the analyte of interest in aqueous and/or non-aqueous liquid
phase samples. Methods have been identified for qualitative and confirmatory determination.
Soil and sediment phase sample methods. The recommended methods/procedures for sample
preparation and analysis to measure the analyte of interest in soil and sediment samples. Methods
have been identified for qualitative and confirmatory determination.
• Surface wipe sample methods. The recommended methods/procedures for sample preparation and
analysis to measure the analyte of interest in surface wipe samples. Methods have been identified for
qualitative and confirmatory determination.
Air filter sample methods. The recommended methods/procedures for sample preparation and
analysis to measure the analyte of interest in air filter samples. Methods have been identified for
qualitative and confirmatory determination.
• Qualitative determination method identifier. A unique identifier or number assigned to an
analytical method by the method publisher. The identified method is intended to determine the
presence of a radiological element or isotope. These methods are less precise than confirmatory
methods, and are used when greater sample throughput and more rapid reporting of results is
required.
Confirmatory method identifier. A unique identifier or number assigned to an analytical method by
the method publisher. The identified method is for measurement of the activity from a particular
radioisotope per unit of mass, volume, or area sampled.
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Section 6 - Selected Radiochemical Methods
6.1 General Guidance
The guidance summarized in this section provides a general overview of how to identify the appropriate
radiochemical method(s) for a given analyte-sample type combination as well as recommendations for
quality control procedures.
For additional information on the properties of the radionuclides listed in Appendix B, TOXNET
Qittgy/toxncyilnijiih.goWindcjyibtil), a cluster of databases on toxicology, hazardous chemicals, and
related areas maintained by the National Library of Medicine, is an excellent resource. EPA's Radiation
Information (http://www.cpa.gov/radiation/radionuclidcs/indcx.html) and the Multi-Agency Radiological
Laboratory Analytical Protocols Manual (MARLAP) (http://www.cpa.gov/radiation/marlap/maiiual.html)
Web sites provide some additional information pertaining to radionuclides of interest and radiochemical
methods.
6.1.1 Standard Operating Procedures for Identifying Radiochemical Methods
To determine the appropriate method that is to be used on an environmental sample, locate the analyte of
concern in Appendix B: Radiochemical Methods under the "Analyte Class" or "Analyte(s)" column.
After locating the analyte of concern, continue across the table to identify the appropriate determinative
technique (e.g., alpha or beta spectrometry), then identify the appropriate qualitative and/or confirmatory
method for the sample type of interest (drinking water, aqueous and liquid phase, soil and sediment,
surface wipes, and air filters) for the particular analyte.
Sections 6.2.1 through 6.2.27 below provide summaries of the qualitative and confirmatory methods
listed in Appendix B. Once a method has been identified in Appendix B, Table 6-1 can be used to locate
the method summary.
Table 6-1. Radiochemical Methods and Corresponding Text Section Numbers
Analyte / Analyte Class
Gross Alpha
Gross Beta
Gamma
Americium-241
Californium-252
CASRN
NA
NA
NA
14596-10-2
13981-17-4
Method
900.0 (EPA)
FRMAC, Vol 2, pg. 33
AP1 (ORISE)
7110 B(SM)
901.1 (EPA)
Ga-01-R (HASL-300)
Am-01-RC(HASL-300)
Am-02-RC (HASL-300)
Am-04-RC (HASL-300)
Pu-12-RC (HASL-300)
AP11 (ORISE)
D3084 (ASTM)
Am-01-RC (HASL-300)
Am-04-RC (HASL-300)
Pu-12-RC (HASL-300)
AP11 (ORISE)
D3084 (ASTM)
Section
6.2.2
6.2.16
6.2.17
6.2.21
6.2.3
6.2.12
6.2.9
6.2.10
6.2.11
6.2.14
6.2.18
6.2.19
6.2.9
6.2.11
6.2.14
6.2.18
6.2.19
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Section 6 - Selected Radiochemical Methods
Analyte / Analyte Class
Cesium-137
Cobalt-60
Curium-244
Europium-154
lridium-192
Plutonium-238
Plutonium-239
Polonium-210
Radium-226
Ruthenium-103
Ruthenium-106
Selenium-75
Strontium-90
Uranium-238
CASRN
10045-97-3
10198-40-0
13981-15-2
15585-10-1
14694-69-0
13981-16-3
15117-48-3
13981-52-7
13982-63-3
13968-53-1
13967-48-1
14265-71-5
10098-97-2
7440-61-1
Method
901.1 (EPA)
Ga-01-R (HASL-300)
7120 (SM)
Am-01-RC (HASL-300)
Am-04-RC (HASL-300)
Pu-12-RC (HASL-300)
AP11 (ORISE)
D3084 (ASTM)
901.1 (EPA)
Ga-01-R (HASL-300)
7120 (SM)
EMSL-33 (EPA)
AP11 (ORISE)
D3084 (ASTM)
Method 111 (EPA)
Po-02-RC (HASL-300)
903.0 (EPA)
903.1 (EPA)
EMSL-19(EPA)
D3084 (ASTM)
7500-Ra B (SM)
7500-Ra C (SM)
901.1 (EPA)
Ga-01-R (HASL-300)
7120 (SM)
Sr-03-RC (HASL-300)
7500-SrB(SM)
908.0 (EPA)
EMSL-33 (EPA)
AP11 (ORISE)
D3084 (ASTM)
D3972 (ASTM)
7500-U B (SM)
7500-U C (SM)
Section
6.2.3
6.2.12
6.2.22
6.2.9
6.2.11
6.2.14
6.2.18
6.2.19
6.2.3
6.2.12
6.2.22
6.2.8
6.2.18
6.2.19
6.2.1
6.2.13
6.2.4
6.2.5
6.2.7
6.2.19
6.2.23
6.2.24
6.2.3
6.2.12
6.2.22
6.2.15
6.2.25
6.2.6
6.2.8
6.2.18
6.2.19
6.2.20
6.2.26
6.2.27
The method summaries are listed in order of method selection hierarchy (see Figure 2-1), starting with
EPA methods followed by methods from other federal agencies and voluntary consensus standard bodies
(VCSB). Methods are listed in numerical order under each publisher. Where available, a direct link to
the full text of the selected analytical method is provided in the method summary. For additional
information regarding sample preparation and analysis procedures and on methods available through
consensus standards organizations, please use the contact information provided in Table 6-2.
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Section 6 - Selected Radiochemical Methods
Table 6-2. Sources of Radiochemical Methods
Name
Publisher
Reference
National Environmental
Methods Index (NEMI)
U.S. Environmental Protection
Agency (USEPA), United States
Geological Survey (USGS)
http^/wwwjTemLgoy
CFR Promulgated Test
Methods (TM)
U.S. Environmental Protection
Agency (USEPA),
Technical Transfer Network (TTN)
Emission Measurement Center
(EMC)
^^
Prescribed Procedures for
Measurement of Radioactivity
in Drinking Water (EPA-600 4-
80-032, August 1980)
U.S. Environmental Protection
Agency (USEPA), Office of
Research and Development
(ORD), Environmental Monitoring
and Support Laboratory (EMSL)
Available from National Technical
Information Service (NTIS)*. NTIS, U.S.
Department of Commerce, 5285 Port
Royal Road, Springfield, VA 22161, (703)
605-6000.
Radiochemical Analytical
Procedures for Analysis of
Environmental Samples, March
1978. EMSL-LV-0539-17
United States Environmental
Protection Agency (USEPA)
Environmental Monitoring and
Support Laboratory (EMSL)
Available from National Technical
Information Service (NTIS)* NTIS, U.S.
Department of Commerce, 5285 Port
Royal Road, Springfield, VA 22161, (703)
605-6000.
EML Procedures Manual,
HASL-300, 28th Edition,
February, 1997
U.S. Department of Energy (DOE),
Environmental Measurements
Laboratory (EML) / Now, U.S.
Department of Homeland Security
(DHS)
s^q^^
cmarLcfm
Also available from National Technical
Information Service (NTIS)* NTIS, U.S.
Department of Commerce, 5285 Port
Royal Road, Springfield, VA 22161, (703)
605-6000.
Federal Radiological Monitoring
and Assessment Center
(FRMAC) Laboratory Manual
United States Department of
Energy (DOE)
National Nuclear Security
Administration (NNSA)
http://www.nv.doe.gov/nationalsecurity/ho
melandsecuritv/frmac/manuals.aspx
Oak Ridge Institute for Science
and Education (ORISE)
Laboratory Procedures Manual
Oak Ridge Institute of Science and
Education (ORISE)
Independent Environmental
Assessment and Verification
http://orise.orau.gov/ieav/survey-
proiects/lab-manual.htm
Annual Book of ASTM
Standards, Vol. 11.02*
American Society for Testing and
Materials (ASTM) International
http://www.astm.org
Standard Methods for the
Examination of Water and
Wastewater, 21st Edition, 2005*
American Public Health
Association (APHA), American
Water Works Association
(AWWA), and Water Environment
Federation (WEF)
http://www.standardmethods.org
' Subscription and/or purchase required.
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Section 6 - Selected Radiochemical Methods
6.1.2 General Quality Control (QC) Guidance for Radiochemical Methods
Having data of known and documented quality is critical for public officials to assess accurately the
activities that may be needed in responding to emergency situations. Having such data requires that
laboratories: (1) conduct the necessary QC to ensure that measurement systems are in control and
operating correctly, (2) properly document results of the analyses, and (3) properly document
measurement system evaluation of the analysis-specific QC. Ensuring data quality also requires that
laboratory results are properly evaluated and the results of the data quality evaluation are transmitted to
decision makers.
The level or amount of QC needed often depends on the intended purpose of the data that are generated.
Various levels of QC may be required if the data are generated during contaminant presence/absence
qualitative determinations versus confirmatory analyses. The specific needs for data generation should be
identified. Quality control requirements and data quality objectives should be derived based on those
needs, and should be applied consistently across laboratories when multiple laboratories are used. For
example, during rapid sample screening analyses, minimal QC samples (e.g., blanks, duplicates) and
documentation might be required to ensure data quality. Implementation of the analytical methods for
evaluation of environmental samples during site assessment through site clearance, such as those
identified in this document, might require increased QC.
Some method-specific QC requirements are described in many of the individual methods that are cited in
this manual. QC requirements will be referenced in standardized analytical protocols developed to
address specific analytes and sample types of concern. Individual methods, sampling and analysis
protocols, or contractual statements of work also should be consulted to determine any additional QC that
may be needed.
QC samples are required to assess the precision, bias, and reliability of sample results. All QC results are
tracked on control charts and reviewed for acceptability and trends in analysis or instrument operation.
Quality control parameters are measured as required per method at the prescribed frequency. QC of
laboratory analyses using radiochemical methods includes ongoing analysis of QC samples and tracking
QC parameters including, but not limited to the following:
• Method blanks;
• Calibration checks;
• Sample and sample duplicates;
• Laboratory control sample recoveries for samples that are not chemically prepared; or
• Matrix spike and matrix spike duplicate recoveries for samples that are chemically prepared; and
Tracer and/or carrier yield.
Please note: The appropriate point of contact identified in Section 4 should be consulted regarding
appropriate QA/QC procedures prior to sample analysis. These contacts will consult with the EPA
coordinator responsible for laboratory activities during the specific event to ensure QA/QC procedures are
performed consistently across laboratories. EPA program offices will be responsible for ensuring that the
QA/QC practices are implemented.
6.1.3 Safety and Waste Management
It is imperative that safety precautions be used during collection, processing, and analysis of
environmental samples. Laboratories should have a documented health and safety plan for handling
samples that may contain target chemical, biological, or radiological contaminants, and laboratory staff
should be trained in and implement the safety procedures included in the plan. In addition, many of the
methods summarized or cited in Section 6.2 contain specific requirements, guidance, or information
regarding safety precautions that should be followed when handling or processing environmental samples
SAM Revision 3.1 89 November 15, 2007
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Section 6 - Selected Radiochemical Methods
and reagents. These methods also provide information regarding waste management. Other resources
that can be consulted for additional information include the following:
• Occupational Health and Safety Administration's Standard for Occupational Exposure to Hazardous
Chemicals in Laboratories (29 CFR 1910.1450)
. Environmental Protection Agency's Standards Regulating Hazardous Waste (40 CFR parts 260-270)
• Standards for Protection Against Radiation (10 CFR part 20)
. U.S. Department of Energy (DOE). Order O 435.1: Radioactive Waste Management. July 1, 1999.
Available at: wj^w-dircy^tivj^yj^^
U.S. Department of Energy (DOE). M 435.1-1. Radioactive Waste Management Manual . Office of
Environmental Management. July 9, 1999. Available at:
U.S. Department of Energy (DOE). Compendium of EPA-Approved Analytical Methods for
Measuring Radionuclides in Drinking Water. Prepared by the Office of Environmental Policy and
Assistance Air, Water and Radiation Division (EH-412). June 1998
U.S. Environmental Protection Agency (EPA). 1996. Profile and Management Options for EPA
Laboratory Generated Mixed Waste. Office of Radiation and Indoor Air, Washington, DC
EPA 402-R-96-015. August. Profile and Management Options for EPA Laboratory Generated
Mixed Waste. Available at: Mtg^7e£a^gOT/radiation/dw
• U.S. Environmental Protection Agency (EPA). 2001. Changes to 40 CFR 266 (Storage, Treatment,
Transportation, and Disposal of Mixed Waste), Federal Register 66:27217-27266, May 16
• U.S. Environmental Protection Agency (EPA). 2002. RCRA Orientation Manual. Office of Solid
Waste, Washington, DC. EPA530-R-02-016. 259 pp. Available at:
http://www.epa.gov/epaoswer/general/orientat/
• Waste Management in a Radioanalytical Laboratory, Chapter 17 Multi -Agency Radiological
Laboratory Analytical Protocols (MARLAP) Manual, July 2004
• National Research Council. 1995. Prudent Practices in the Laboratory; Handling and Disposal of
Chemicals, National Academy Press, Washington, DC
• National Council on Radiation Protection and Measurements (NCRP). 2002. Risk-Based
Classification of Radioactive and Hazardous Chemical Wastes, 7910 Woodmont Avenue, Suite 400,
Bethesda, MD 208 14-3095
• U.S. Nuclear Regulatory Commission/U.S. Environmental Protection Agency (NRC/EPA). 1995.
Low-Level Mixed Waste Storage Guidance, Federal Register 60:40204-4021 1, August 7
6.2 Method Summaries
Summaries for the analytical methods listed in Appendix B are provided in Sections 6.2.1 through 6.2.27 '.
These summaries contain information that has been extracted from the selected methods. Each method
summary contains a table identifying the contaminants in Appendix B to which the method applies, a
brief description of the analytical method, and a link to the full version of the method or a source for
obtaining a full version of the method. The full version of the method should be consulted prior to
sample analysis.
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Section 6 - Selected Radiochemical Methods
6.2.1 EPA Method 111: Determination of Polonium-210 Emissions from Stationary
Sources
This method should be used for qualitative and confirmatory analysis of surface wipes and air filters for
the contaminant identified below and listed in Appendix B.
Analyte(s)
Polonium-210
CASRN
13981-52-7
This method covers the determination of polonium-210 in particulate matter samples collected from
stationary sources such as exhaust stacks. Polonium-210 in the sample is put in solution, deposited on a
metal disc, and the radioactive disintegration rate measured. Polonium in acid solution spontaneously
deposits on surface metals that are more electropositive than polonium.
Source: 40 CFR 61 Appendix B; National Emission Standards for Air Pollutants; Appendix B to Part 61 -
Test Methods, U.S. Environmental Protection Agency (EPA). Emission Measurement Center (EMC).
February 2000. Prepared by the Office of Air Quality Planning and Standards (OAQPS), Research
Triangle Park, North Carolina, 27711. Also at: http://www.epa.gov/ttn/emc/promgate.html
6.2.2 EPA Method 900.0: Gross Alpha and Gross Beta Radioactivity in Drinking Water
This method should be used for gross alpha and gross beta determination in drinking water samples.
The method provides an indication of the presence of alpha and beta emitters, including the following
SAM analytes:
• Americium-241 (CAS RN 14596-10-2) Alpha emitter
• Californium-252 (CAS RN 13981-17-4) Alpha emitter
• Cesium-137 (CAS RN 10045-97-3) Beta emitter
• Cobalt-60 (CASRN 10198-40-0) Beta emitter
• Curium-244 (CAS RN 13981-15-2) Alpha emitter
• Europium-154 (CAS RN 15585-10-1) Beta emitter
• Iridium-192 (CAS RN 14694-69-0) Beta emitter
• Plutonium-238 (CAS RN 13981-16-3) Alpha emitter
• Plutonium-239 (CAS RN 15117-48-3) Alpha emitter
• Polonium-210 (CAS RN 13981-52-7) Alpha emitter
• Radium-226 (CAS RN 13982-63-3) Alpha emitter
• Ruthenium-103 (CAS RN 13968-53-1) Beta emitter
• Ruthenium-106 (CAS RN 13967-48-1) Beta emitter
• Strontium-90 (CAS RN 10098-97-2) Beta emitter
• Uranium-238 (CAS RN 7440-16-1) Alpha emitter
An aliquot of a preserved drinking water sample is evaporated to a small volume (3 to 5 mL) and
transferred quantitatively to a tared 2-inch planchet. The aliquot volume is determined based on a
maximum total solids content of 100 mg. The sample aliquot is evaporated to dryness in the planchet to a
constant weight, cooled, and counted using a gas proportional or scintillation counting system. The
counting system is calibrated with thorium-230 for gross alpha, or with cesium-137 for gross beta
analysis. A traceable standards-based absorption curve must be developed for each calibration nuclide
(Th-230 or Cs-137) based on a range of total solids content in the 2-inch planchet from 0 to 100 mg. The
results are corrected for the absorption factor based total solids on the planchet.
Source: Prescribed Procedures for Measurement of Radioactivity in Drinking Water, National Exposure
Risk Laboratory-Cincinnati (NERL-CI), EPA/600/4/80/032, August 1980, available from National
Technical Information Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161. Phone: 800-553-
6847. Also at
http://infotrek.er.usgs.gov/pls/apex/f?p=119:38:1908830694668232::::P38_METHOD_ID:4730
SAM Revision 3.1 91 November 15, 2007
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6.2.3 EPA Method 901.1: Gamma Emitting Radionuclides in Drinking Water
This method should be used for qualitative and confirmatory analysis of drinking water samples for the
contaminants identified below and listed in Appendix B.
Analyte(s)
Cesium-1371
Cobalt-60
Europium-154
lridium-192
Ruthenium-103
Ruthenium-1061
Selenium-75
CASRN
10045-97-3
10198-40-0
15585-10-1
14694-69-0
13968-53-1
13967-48-1
14265-71-5
1 The method will measure decay products of these isotopes
This method is applicable for analysis of water samples that contain radionuclides that emit gamma
photons with energies ranging from approximately 60 to 2000 keV. The method uses gamma
spectroscopy for measurement of gamma photons emitted from radionuclides without separating them
from the sample matrix. A homogeneous aliquot of water is placed into a standard geometry (normally a
Marinelli beaker) for gamma counting, typically using a High Purity Germanium (HPGe) detector.
Detectors such as Germanium (Lithium) (Ge(Li)) or thallium-activated sodium iodide (Nal(Tl)) also can
be used. Sample aliquots are counted long enough to meet the required sensitivity of measurement. To
reduce adsorbance of radionuclides on the walls of the counting container, the sample is acidified at
collection time. Due to its lower resolution, significant interference can occur using the Nal(Tl) detector
when counting a sample containing radionuclides that emit gamma photons of similar energies. When
using this method, shielding is needed to reduce background interference. Detection limits are dependent
on sample volume, geometry (physical shape), and counting time.
Source: Prescribed Procedures for Measurement of Radioactivity in Drinking Water, National Exposure
Risk Laboratory-Cincinnati (NERL-CI), EPA/600/4/80/032, August 1980, available from National
Technical Information Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161. Phone: 800-553-
6847.
6.2.4 EPA Method 903.0: Alpha-Emitting Radium Isotopes in Drinking Water
This method should be used for qualitative determination in drinking water samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Radium-2261
CASRN
13982-63-3
1 The method will measure decay products of these isotopes
This method covers measurement of the total soluble alpha emitting radioisotopes of radium, namely
radium-223, radium-224 and radium-226 in drinking water. The method does not give an accurate
measurement of radium-226 content in the sample when other alpha emitters are present. If radium-223
and radium-224 are present, the results can be used to provide a gross determination of radium-226. When
the total radium alpha activity of a drinking water sample is greater than 5 pCi/L, use of Method 903.1
(Radium-226 in Drinking Water) is preferred. Radium in the water sample is collected by coprecipitation
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with barium and lead sulfate, and purified by re-precipitation from ethylenediaminetetraacetic acid
(EDTA) solution. Citric acid is added to ensure that complete interchange occurs before the first
precipitation step. The final barium sulfate precipitate is alpha counted to determine the total
disintegration rate of the radium isotopes. By making a correction for the ingrowth of radon and its alpha
emitting progeny for the elapsed time after separation, one can determine radium activity in the sample.
Presence of significant natural barium in the sample can result in a falsely high yield. Based on a 1000-
mL sample and 100-minute counting time, the minimum detectable level for this method is 0.5 pCi/L.
Source: Prescribed Procedures for Measurement of Radioactivity in Drinking Water, National Exposure
Risk Laboratory-Cincinnati (NERL-CI), EPA/600/4/80/032, August 1980, available from National
Technical Information Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161. Phone: 800-553-
6847. Also at
http://infotrek.er.usgs.gov/pls/apex/f?p=119:38:1908830694668232::::P38_METHOD_ID:9183
6.2.5 EPA Method 903.1: Radium-226 in Drinking Water - Radon Emanation Technique
This method should be used for confirmatory analysis of drinking water samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Radium-2261
CASRN
13982-63-3
1 The method will measure decay products of these isotopes
This method is specific for radium-226, and is based on the emanation and scintillation counting of radon-
222, a daughter product of radium-226. Radium-226 is concentrated and separated from the water sample
by coprecipitation on barium sulfate. The precipitate is dissolved in EDTA reagent, placed in a sealed
bubbler and stored for ingrowth of radon-222. After ingrowth, the gas is purged into a scintillation cell.
When the short-lived radon-222 daughters are in equilibrium with the parent (after ~4h), the scintillation
cell is counted for activity. The absolute measurement of radium-226 is effected by calibrating the
scintillation cell system with a standard solution of the nuclide. There are no radioactive interferences in
this method. Based on a 1000-mL sample and 100-minute counting time, the minimum detectable level
for this method is 0.5 pCi/L.
Source: Prescribed Procedures for Measurement of Radioactivity in Drinking Water, National Exposure
Risk Laboratory-Cincinnati (NERL-CI), EPA/600/4/80/032, August 1980, available from National
Technical Information Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161. Phone: 800-553-
6847. Also at
6.2.6 EPA Method 908.0: Uranium in Drinking Water - Radiochemical Method
This method should be used for qualitative determination in drinking water samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Uranium-2381
CASRN
7440-61-1
11f it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
This method measures total uranium alpha activity of a sample, without doing an isotopic uranium
analysis. The sample is acidified with hydrochloric acid and boiled to eliminate carbonate and
bicarbonate ions. Uranium is coprecipitated with ferric hydroxide and separated from the sample. The
uranium is then separated from other radionuclides that were carried down with the ferric hydroxide by
dissolving the hydroxide precipitate in hydrochloric acid, putting the solution through an anion exchange
SAM Revision 3.1 93 November 15, 2007
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Section 6 - Selected Radiochemical Methods
column, washing the column with hydrochloric acid, and finally eluting the uranium with hydrochloric
acid. The uranium eluate is evaporated and the uranium chemical form is converted to nitrate. The
residue is transferred to a stainless steel planchet, dried, flamed, and counted for alpha particle activity.
Since uranium is a naturally occurring radionuclide, reagents must be checked for uranium contamination
by analyzing a complete reagent blank by the same procedure as used for the samples. Based on a 1000-
mL sample and 100-minute counting time in a single laboratory study, the minimum detectable level for
this method is 1.0 pCi/L.
Source: Prescribed Procedures for Measurement of Radioactivity in Drinking Water, National Exposure
Risk Laboratory-Cincinnati (NERL-CI), EPA/600/4/80/032, August 1980, available from National
Technical Information Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161. Phone: 800-553-
6847. Also at
http://infotrek.er.usgs.gov/pls/apex/f?p=119:38:1908830694668232::::P38_METHOD_ID:9184
6.2.7 EPA Method EMSL-19: Determination of Radium-226 and Radium-228 in Water,
Soil, Air and Biological Tissue
This method should be used for confirmatory analysis of soil/sediment, surface wipe, and air filter
samples for the contaminant identified below and listed in Appendix B.
Analyte(s)
Radium-2261
CASRN
13982-63-3
1 The method will measure decay products of these isotopes
Following acid digestion and filtration of soil, sediment, surface wipe, or air filter samples, radium is
precipitated with barium sulfate. Barium-radium-sulfate is dissolved in a pentasodium
diethylenetriamine-pentaacetate solution and transferred to an emanation tube. The radon is allowed to
come to equilibrium for approximately 30 days. Radium-226 decays by alpha emission to radon-222.
Radon-222 is separated and collected from the liquid by a de-emanation technique. The radon is counted
by alpha scintillation 4.5 hours after de-emanation, at which time the short-lived progeny have reached
97% of equilibrium. An applicable measurement range has not been determined; however, samples that
contain 0.1 pCi of Radium-226 have been analyzed.
Source: Radiochemical Analytical Procedures for Analysis of Environmental Samples, United States
Environmental Protection Agency, Environmental Monitoring and Support Laboratory (EMSL), March
1979, available from National Technical Information Service (NTIS), 5285 Port Royal Road, Springfield,
VA 22161. Phone:800-553-6847.
6.2.8 EPA Method EMSL-33: Isotopic Determination of Plutonium, Uranium, and
Thorium in Water, Soil, Air, and Biological Tissue
This method should be used for confirmatory analysis of drinking water, aqueous/liquid, soil/sediment,
surface wipe, and/or air filter samples for the contaminants identified below and listed in Appendix B.
Analyte(s)
Plutonium-2381
Plutonium-2391
Uranium-2381
CASRN
13981-16-3
15117-48-3
7440-61-1
11f it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
SAM Revision 3.1 94 November 15, 2007
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Section 6 - Selected Radiochemical Methods
This method is appropriate for the analysis of isotopic plutonium, uranium, and thorium, together or
individually, by alpha spectrometry. Plutonium-236, uranium-232, and thorium-234 tracer standards are
added for the determination of chemical yields. Samples are decomposed by nitric-hydrofluoric acid
digestion or ignition to assure that all of the plutonium is dissolved and chemically separated from the
sample by coprecipitation with sodium and ammonium hydroxide, anion exchange, and electrodeposition.
The residues are dissolved in dilute nitric acid and successive sodium and ammonium hydroxide
precipitations are performed in the presence of boric acid to remove fluoride and soluble salts. The
hydroxide precipitate is dissolved, the solution is pH-adjusted with hydrochloric acid, and plutonium and
uranium are adsorbed on an anion exchange column, separating them from thorium. Plutonium is eluted
with hydrobromic acid. The actinides are electrodeposited on stainless steel discs from an ammonium
sulfate solution and subsequently counted by alpha spectrometry. This method is designed to detect
environmental levels of activity as low as 0.02 pCi per sample. To avoid possible cross-contamination,
sample aliquot activities should be limited to 25 pCi or less.
Source: Radiochemical Analytical Procedures for Analysis of Environmental Samples, United States
Environmental Protection Agency, Environmental Monitoring and Support Laboratory (EMSL), March
1979, available from National Technical Information Service (NTIS), 5285 Port Royal Road, Springfield,
VA 22161. Phone:800-553-6847.
6.2.9 EML HASL-300 Method Am-01-RC: Americium in Soil
This method should be used for confirmatory analysis of soil/sediment samples for the contaminants
identified below and listed in Appendix B.
Analyte(s)
Americium-2411
Californium-2521
Curium-2441
CASRN
14596-10-2
13981-17-4
13981-15-2
1lf it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
This method uses alpha spectrometry for determination of americium-241 in soil, and also can be applied
for determination of californium. Americium is leached from soil with nitric acid and hydrochloric acid.
Americium-243 is added as a tracer to determine chemical yield. The soil is processed through the
plutonium separation steps using ion exchange resin according to Method Pu-11-RC. Americium is
collected with a calcium oxalate precipitation and finally isolated and purified by ion exchange.
Californium-252 and curium-244 are eluted with americium as americium is stripped off the column.
After source preparation by microprecipitation, americium-241, californium-252, and curium-244 are
determined by alpha spectrometry analysis. The counting period chosen depends on the sensitivity
required of the measurement and the degree of uncertainty in the result that is acceptable. The lower limit
of detection (LLD) for americium-241 is 0.5 mBq when counted for 1000 minutes. In cases where less
than 100 g of sample is available, use of Pu-12-RC is recommended.
Source: "Am-01-RC, Pu-11-RC, and Pu-12-RC." EML Procedures Manual, HASL-300, 28th Edition,
Environmental Measurements Laboratory (EML), Department of Energy (EML is currently part of the
U.S. Department of Homeland Security), February 1997. Web:
SAM Revision 3.1 95 November 15, 2007
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Section 6 - Selected Radiochemical Methods
6.2.10 EML HASL-300 Method Am-02-RC: Americium-241 in Soil-Gamma Spectrometry
This method should be used for qualitative determination in soil/sediment samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Americium-2411
CASRN
14596-10-2
1lf it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
This method uses gamma spectrometry for determination of americium-241 in soil. Americium-241
decays with the emission of a gamma ray at 59.5 keV with a decay frequency (abundance or yield) of
35.9%. The sample is placed into an appropriately sized standard geometry (normally a Marinelli beaker)
after drying and grinding the sample for homogenization. Gamma-ray attenuation corrections are
required if the calibration source and the sample are in a different matrix or are of different densities. The
lower limit of detection (LLD) for 600 to 800 g of soil in a Marinelli beaker is 0.74 mBq for a 1000-
minute count.
Source: EML Procedures Manual, HASL-300, 28th Edition, Environmental Measurements Laboratory
(EML), Department of Energy (EML is currently part of the U.S. Department of Homeland Security),
February 1997. Web: http://www.cml.st.dhs.gov/publications/procman.cfm
6.2.11 EML HASL-300 Method Am-04-RC: Americium in QAP Water and Air Filters -
Eichrom's TRU Resin
This method should be used for confirmatory analysis of drinking water and aqueous/liquid samples for
the contaminants identified below and listed in Appendix B.
Analyte(s)
Americium-2411
Californium-2521
Curium-2441
CASRN
14596-10-2
13981-17-4
13981-15-2
1lf it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
This method is specific to measurement of americium isotopes in samples that do not contain lanthanides,
but also can be used for measurement of californium and curium. The method uses microprecipitation
and determination by alpha spectrometry. Americium-243 is added to the sample to determine chemical
yield. The sample is processed through separation steps using ion exchange resins. The eluate from the
ion exchange column containing americium (and all other ions, except plutonium) is evaporated,
redissolved, and loaded onto a Transuranic (TRU) Resin extraction column. Americium (and curium and
californium, if present) is separated and purified on the column and finally stripped with dilute nitric acid
stripping solution. Microprecipitation is used to prepare for alpha spectrometry. The method involves
sample preparation steps from EML HASL-300 Method Pu-10-RC for water samples. The lower limit of
detection (LLD) for total americium is 0.3 mBq when counted for 1000 minutes.
Source: EML Procedures Manual, HASL-300, 28th Edition, Environmental Measurements Laboratory
(EML), Department of Energy (EML is currently part of the U.S. Department of Homeland Security),
February 1997. Web: httPI/Avrnvjatiljitd^^
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Section 6 - Selected Radiochemical Methods
6.2.12 EML HASL-300 Method Ga-01-R: Gamma Radioassay
This method should be used for qualitative and/or confirmatory analysis of soil/sediment, surface
wipes, and/or air filter samples for the contaminants identified below and listed in Appendix B.
Analyte(s)
Cesium-1371
Cobalt-60
Europium-154
lridium-192
Ruthenium-103
Ruthenium-1061
Selenium-75
CASRN
10045-97-3
10198-40-0
15585-10-1
14694-69-0
13968-53-1
13967-48-1
14265-71-5
1 The method will measure decay products of these isotopes
This method uses gamma spectroscopy for the measurement of gamma photons emitted from
radionuclides without separating them from the sample matrix. Samples are placed into a standard
geometry for gamma counting, typically using a High Purity Germanium (HPGe) detector. Detectors
such as Germanium (Lithuim) (Ge(Li)) or thallium-activated sodium iodide (Nal(Tl)) also can be used.
The sample is placed into a standard geometry (physical shape) for gamma counting. Soil samples and
sludge are placed into an appropriately sized Marinelli beaker after drying and grinding the sample for
homogenization. Air filters and surface wipes can be counted directly or pressed into a planchet and
counted. Samples are counted long enough to meet the required sensitivity of measurement. For typical
counting systems and sample types, activity levels of approximately 40 Bq are measured, and sensitivities
as low as 0.002 Bq can be achieved for many nuclides. Because of electronic limitations, count rates
higher than 2000 counts per second (cps) should be avoided. High activity samples may be diluted,
reduced in size, or moved away from the detector (a limited distance) to reduce the count rate and allow
for analysis. The method is applicable for analysis of samples that contain radionuclides emitting gamma
photons with energies ranging from approximately >40 keV for Germanium (Lithium) (Ge(Li)) and 100
keV for thallium-activated sodium iodide (Nal(Tl)) detectors.
Source: EML Procedures Manual, HASL-300, 28th Edition, Environmental Measurements Laboratory
(EML), Department of Energy (EML is currently part of the U.S. Department of Homeland Security),
February, 1997. Web: http://www.eml.st.dhs.gov/publications/procman.cfm
6.2.13 EML HASL-300 Method Po-02-RC: Polonium in Water, Vegetation, Soil, and Air
Filters
This method should be used for qualitative and confirmatory analysis of drinking water, aqueous/liquid
phases and soil/sediment samples for the contaminant identified below and listed in Appendix B.
Analyte(s)
Polonium-210
CASRN
1-3981-52-7
This method uses alpha spectrometry for determination of polonium in water, vegetation, soil, and air
filter samples. Polonium equilibrated with Po-208 or Po-209 tracer is isolated from most other elements
by coprecipitation with lead sulfide. The sulfide precipitate is dissolved in weak hydrochloric acid
solution. Polonium is quantitatively deposited on a nickel disc, and the plated disc is counted on an alpha
spectrometer to measure chemical yield and activity of the sample. The solution from the deposition may
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Section 6 - Selected Radiochemical Methods
be retained and analyzed for Pb-210. When counted for 1000 minutes, the lower level of detection (LLD)
for polonium is 1.0 mBq for water and 1.3 mBq for vegetation, soil and filters.
Source: EML Procedures Manual, HASL-300, 28th Edition, Environmental Measurements Laboratory
(EML), Department of Energy (EML is currently part of the U.S. Department of Homeland Security),
February, 1997. Web: httpV/wwuv^mj±st^hjIgav/nu^lications/progmaiv.cfin
6.2.14 EML HASL-300 Method Pu-12-RC: Plutonium and/or Americium in Soil or
Sediments
This method is not currently listed for any of the analyte/sample type combinations included in Appendix
B. In cases where only small sample volumes (<100 g) will be analyzed, this method could be used for
confirmatory analysis of soil/sediment samples for the contaminants identified below and listed in
Appendix B.
Analyte(s)
Americium-2411
Californium-2521
Curium-2441
CASRN
14596-10-2
13981-17-4
13981-15-2
1lf it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
A sample of soil of up to 100 g in size is equilibrated with Am-243 tracer. Contaminant isotopes are
leached with nitric and hydrochloric acid. Plutonium is removed by ion exchange. The eluent from the
plutonium separation is saved for determination of americium, curium, and californium. Americium,
curium, and californium are collected with a calcium oxalate coprecipitation, isolated and purified by
extraction chromatography. Microprecipitation is used to prepare the sample for analysis by alpha
spectrometry of americium, curium, and californium. The lower limit of detection for Americium is 0.5
mBq when counted for 1000 minutes.
Source: EML Procedures Manual, HASL-300, 28th Edition, Environmental Measurements Laboratory
(EML), Department of Energy (EML is currently part of the U.S. Department of Homeland Security),
February, 1997. Web: http://www.cml.st.dhs.gov/publications/procman.cfin
6.2.15 EML HASL-300 Method Sr-03-RC: Strontium-90 in Environmental Samples
This method should be used for qualitative and confirmatory analysis of soil/sediment, surface wipes,
and air filter samples for the contaminant identified below and listed in Appendix B.
Analyte(s)
Strontium-901
CASRN
10098-97-2
1 The method will measure decay products of these isotopes
Strontium is separated from calcium, other fission products, and natural radioactive elements. Fuming
nitric acid separations remove the calcium and most other interfering ions. Radium, lead and barium are
removed with barium chromate. Traces of other fission products are scavenged with iron hydroxide.
After strontium-90 and yttrium-90 equilibrium has been attained, yttrium-90 is precipitated as the
hydroxide and converted to oxalate for counting on a low-background gas proportional beta counter.
Chemical yield is determined with strontium-85 tracer by counting in a gamma well detector.
SAM Revision 3.1 98 November 15, 2007
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Section 6 - Selected Radiochemical Methods
Source: EML Procedures Manual, HASL-300, 28th Edition, Environmental Measurements Laboratory
(EML), Department of Energy (EML is currently part of the U.S. Department of Homeland Security),
February, 1997. Web: http://www.cml.st.dhs.gov/publications/procman.cfin
6.2.16 FRMAC Method Volume 2, Page 33: Gross Alpha and Beta in Air
This method should be used for gross alpha and gross beta determination in air filters, and can also be
used as a direct counting of surface wipes. The method provides an indication of the presence of alpha
and beta emitters, including the following SAM analytes:
Americium-241 (CAS RN 14596-10-2) Alpha emitter
Californium-252 (CAS RN 13981-17-4) Alpha emitter
Cesium-137 (CAS RN 10045-97-3) Beta emitter
Cobalt-60 (CAS RN 10198-40-0) Beta emitter
Curium-244 (CAS RN 13981-15-2) Alpha emitter
Europium-154 (CAS RN 15585-10-1) Beta emitter
Iridium-192 (CAS RN 14694-69-0) Beta emitter
Plutonium-238 (CAS RN 13981-16-3) Alpha emitter
Plutonium-239 (CAS RN 15117-48-3) Alpha emitter
Polonium-210 (CAS RN 13981-52-7) Alpha emitter
Radium-226 (CAS RN 13982-63-3) Alpha emitter
Ruthenium-103 (CAS RN 13968-53-1) Beta emitter
Ruthenium-106 (CAS RN 13967-48-1) Beta emitter
Strontium-90 (CAS RN 10098-97-2) Beta emitter
Uranium-238 (CAS RN 7440-16-1) Alpha emitter
This method allows for measurement of gross alpha and gross beta radiation in air samples. The method
also can be applied for the analysis of surface swipes. A thin-window gas-flow proportional counter is
used for counting gross alpha and beta radioactivity. The method supplies an approximation of the alpha
and beta activity present in the air or the removable surface activity dependent on the sample type. For
this application, the procedure requires the use of thorium-230 for alpha counting efficiency and cesium-
137 for beta counting efficiency in the calibration of the detector. An air filter or swipe sample is placed
onto a planchet, then counted for alpha and beta radioactivity. Activity is reported in activity units per
volume of air sampled, as units of activity per surface area sampled, or as total units of activity in cases
where sample collection information is not available.
Source: FRMAC Monitoring and Analysis Manual - Sample Preparation and Analysis - Volume 2,
Federal Radiological Monitoring and Assessment Center (FRMAC), DOE/NV/1178-181 Vol. 2, UC-707,
August 1998. Web: http://www.nv.doc.gov/nationalsccurity/homclandsccurity/frmac/manuals.aspx
6.2.17 ORISE Method AP-1: Gross Alpha and Beta for Various Matrices
This method should be used for gross alpha and gross beta determination in soil/sediment samples for
the contaminants identified below and listed in Appendix B. This method provides an indication of the
presence of alpha and beta emitters, including the following SAM analytes:
• Americium-241 (CAS RN 14596-10-2) Alpha emitter
• Californium-252 (CAS RN 13981-17-4) Alpha emitter
• Cesium-137 (CAS RN 10045-97-3) Beta emitter
• Cobalt-60 (CAS RN 10198-40-0) Beta emitter
• Curium-244 (CAS RN 13981-15-2) Alpha emitter
• Europium-154 (CAS RN 15585-10-1) Beta emitter
• Iridium-192 (CAS RN 14694-69-0) Beta emitter
• Plutonium-238 (CAS RN 13981-16-3) Alpha emitter
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• Plutonium-239 (CAS RN 15117-48-3) Alpha emitter
• Polonium-210 (CAS RN 13981-52-7) Alpha emitter
• Radium-226 (CAS RN 13982-63-3) Alpha emitter
• Ruthenium-103 (CAS RN 13968-53-1) Beta emitter
• Ruthenium-106 (CAS RN 13967-48-1) Beta emitter
• Strontium-90 (CAS RN 10098-97-2) Beta emitter
• Uranium-238 (CAS RN 7440-16-1) Alpha emitter
This method covers the measurement of gross alpha and gross beta in various sample types. This
procedure provides screening measurements to indicate whether specific chemical analyses are required
for water, soil, vegetation, and other solids. Liquid samples are acidified, concentrated, dried in a
planchet, and counted in a low-background proportional counter. Solid samples are dried and processed
to provide homogeneity, and a known quantity is transferred to a planchet and counted in a low-
background proportional counter.
Source: Laboratory Procedures Manual for the Environmental Survey and Site Assessment Program;
May 2001; Oak Ridge Institute for Science and Education (ORISE), Oak Ridge Associated Universities
(ORAU). Web: http://orise.orau.gOv/ieav/survey-projects/pubs/lab-manual/l9ap 1 .pdf
6.2.18 ORISE Method AP-11: Sequential Determination of the Actinides in Environmental
Samples Using Total Sample Dissolution and Extraction
This method is not currently listed for any of the analyte/sample type combinations included in Appendix
B. If it is suspected that a sample exists in a refractory form (i.e., non-digestable or dissolvable material
after normal digestion methods) or if there is a matrix interference problem, this method should be
routinely used for confirmatory analysis and, in the event of refractory radioactive material, should be
used for both qualitative determination and confirmatory analysis of drinking water, aqueous/liquid,
soil/sediment, surface wipes, and air filter samples for the contaminants identified below and listed in
Appendix B.
Analyte(s)
Americium-241
Californium-252
Curium-244
Plutonium-238
Plutonium-239
Uranium-238
CASRN
14596-10-2
13981-17-4
13981-15-2
13981-16-3
15117-48-3
7440-61-1
This method covers the measurement of the actinides americium-241, californium-252, curium-244,
plutonium-238 and -239, and uranium-238 in water, soil, and other solids. The method is used if there is
an indication that the material is in a highly refractory or "non-dissolvable" form. Solid and unfiltered
aqueous samples are completely dissolved by a combination of potassium hydrogen fluoride and
pyrosulfate fusions. Filtered aqueous samples are evaporated to dryness followed by a pyrosulfate fusion.
The fusion cake is dissolved and, for analyses requiring uranium only, two barium sulfate precipitations
are performed and the uranium is separated using EDTA. For all other analyses, one barium sulfate
precipitation is performed and all alpha emitters are coprecipitated on barium sulfate. The barium sulfate
is dissolved and the actinides are separated by extraction chromatography. An optional section is
presented for the separation of americium from the lanthanides. All actinides are coprecipitated on
cerium fluoride and counted with an alpha spectrometer system.
SAM Revision 3.1 100 November 15, 2007
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Section 6 - Selected Radiochemical Methods
Source: Laboratory Procedures Manual for the Environmental Survey and Site Assessment Program;
May 2001; Oak Ridge Institute for Science and Education (ORISE), Oak Ridge Associated Universities
(ORAU). Web: http://orisc.orau.gov/icav/survcy-projccts/pubs/lab-manual/apll.pdf
6.2.19 ASTM Method D3084: Standard Practice for Alpha Spectrometry in Water
This method should be used for qualitative determination analysis of drinking water, aqueous/liquid,
soil and sediment, surface wipes, and air filter samples for the contaminants identified below and listed in
Appendix B.
Analyte(s)
Americium-2411
Californium-2521
Curium-2441
Plutonium-2381
Plutonium-2391
Radium-2262
Uranium-2381
CASRN
14596-10-2
13981-17-4
13981-15-2
13981-16-3
15117-48-3
13982-63-3
7440-61-1
1 If it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
2The method will measure decay products of these isotopes
This standard practice covers the process that is required to obtain well-resolved alpha spectra from water
samples and discusses the associated problems. This practice is typically followed with specific chemical
separations and mounting techniques that are included in referenced methods. A chemical procedure is
required to isolate and purify the radionuclides (see ASTM Methods D3865, Standard Test Method for
Plutonium in Water and D3972, Standard Test Method for Isotopic Uranium in Water by
Radiochemistry), and a radioactive tracer is added to determine yield. A source is prepared by employing
electrodeposition, microprecipitation, or evaporation (depositing the solution onto a stainless steel or
platinum disc). Electrodeposition and microprecipitation are preferred. The source's radioactivity is then
measured in an alpha spectrometer according to manufacturer's operating instructions. The counting
period chosen depends on the sensitivity required of the measurement and the degree of uncertainty in the
result that is acceptable.
Source: Annual Book of ASTM Standards, Vol. 11.02, American Society for Testing and Materials
(ASTM), 1996, ASTM International, 100 Barr Harbor Drive West, Conshohocken, PA 19428. Phone:
610-832-9500. Web: MtEJ//www,,astm,Qrg. Use method number when ordering.
6.2.20 ASTM Method D3972: Standard Test Method for Isotopic Uranium in Water by
Radiochemistry
This method should be used for confirmatory analysis of drinking water samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Uranium-2381
CASRN
7440-61-1
1 If it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
SAM Revision 3.1 101 November 15, 2007
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Section 6 - Selected Radiochemical Methods
This method covers the determination of uranium isotopes in water by means of chemical separations and
alpha spectrometry analysis. Uranium is chemically separated from a water sample by coprecipitation
with ferrous hydroxide followed by anion exchange, and electrodeposition. When suspended matter is
present, an acid dissolution step is added to ensure that all of the uranium dissolves. The sample is
acidified, and uranium-232 is added as an isotopic tracer to determine chemical yield. Uranium is
coprecipitated from the sample with ferrous hydroxide. This precipitate is dissolved in concentrated
hydrochloric acid, or is subjected to acid dissolution with concentrated nitric and hydrofluoric acids, if the
hydrochloric acid fails to dissolve the precipitate. Uranium is separated from other radionuclides by
adsorption on anion exchange resins, followed by elution with hydrochloric acid. The uranium is finally
electrodeposited onto a stainless steel disc and counted using alpha spectrometry.
Source: Annual Book of ASTMStandards, Vol. 11.02, American Society for Testing and Materials
(ASTM), 2002, ASTM International, 100 Barr Harbor Drive West, Conshohocken, PA 19428. Phone:
610-832-9500. Web: http://www. .org. Use method number when ordering.
6.2.21 Standard Method 7110 B: Gross Alpha and Gross Beta Radioactivity (Total,
Suspended, and Dissolved)
This method should be used for gross alpha and gross beta determination in aqueous/liquid samples for
the contaminants identified below and listed in Appendix B. The method provides an indication of the
presence of alpha and beta emitters, including the following SAM analytes:
Americium-241 (CAS RN 14596-10-2) Alpha emitter
Californium-252 (CAS RN 13981-17-4) Alpha emitter
Cesium-137 (CAS RN 10045-97-3) Beta emitter
Cobalt-60 (CAS RN 10198-40-0) Beta emitter
Curium-244 (CAS RN 13981-15-2) Alpha emitter
Europium-154 (CAS RN 15585-10-1) Beta emitter
Iridium-192 (CAS RN 14694-69-0) Beta emitter
Plutonium-238 (CAS RN 13981-16-3) Alpha emitter
Plutonium-239 (CAS RN 15117-48-3) Alpha emitter
Polonium-210 (CAS RN 13981-52-7) Alpha emitter
Radium-226 (CAS RN 13982-63-3) Alpha emitter
Ruthenium-103 (CAS RN 13968-53-1) Beta emitter
Ruthenium-106 (CAS RN 13967-48-1) Beta emitter
Strontium-90 (CAS RN 10098-97-2) Beta emitter
Uranium-238 (CAS RN 7440-16-1) Alpha emitter
This method allows for measurement of gross alpha and gross beta radiation in water samples. This
method recommends using a thin-window gas-flow proportional counter for counting gross alpha and
beta radioactivity. An internal proportional or Geiger counter may also be used. An aliquot of sample is
evaporated to a small volume and transferred to a tared counting pan. The sample residue is dried to
constant weight, cooled, and reweighed to determine dry residue weight, then counted for alpha and beta
radioactivity.
Source: Standard Methods for Examination of Water and Wastewater, 21st Edition, American Public
Health Association (APHA), American Water Works Association (AWWA), and Water Environment
Federation (WEF), 2008. Web: httg^7www.standardmcthods±oig/
SAM Revision 3.1 102 November 15, 2007
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Section 6 - Selected Radiochemical Methods
6.2.22 Standard Method 7120: Gamma-Emitting Radionuclides
This method should be used for qualitative and confirmatory analysis of aqueous/liquid samples for the
contaminants identified below and listed in Appendix B and with gamma photon energies ranging from
approximately 60 to 2000 keV.
Analyte(s)
Cesium-1371
Cobalt-60
Europium-154
lridium-192
Ruthenium-103
Ruthenium-1061
Selenium-75
CASRN
10045-97-3
10198-40-0
15585-10-1
14694-69-0
13968-53-1
13967-48-1
14265-71-5
1 The method will measure decay products of these isotopes
The method uses gamma spectroscopy using either germanium (Ge) detectors or thallium-activated
sodium iodide (Nal(Tl)) crystals for the measurement of gamma photons emitted from radionuclides
present in water. The method can be used for qualitative and confirmatory determinations with Ge
detectors or semi-qualitative and semi-quantitative determinations (using Nal(Tl) detectors). Exact
confirmation using Nal is possible for single nuclides or when the gamma emissions are limited to a few
well-separated energies. A homogeneous water sample is placed into a standard geometry (normally a
Marinelli beaker) for gamma counting. Sample portions are counted long enough to meet the required
sensitivity of measurement. A standard containing a mixture of gamma energies from approximately 100
to 2000 keV is used for energy calibration.
Source: Standard Methods for Examination of Water and Wastewater, 21st Edition, American Public
Health Association (APHA), American Water Works Association (AWWA), and Water Environment
Federation (WEF), 2005. Web: http://www.standardmcthods.org/
6.2.23 Standard Method 7500-Ra B: Radium: Precipitation Method
This method should be used for qualitative determination in aqueous/liquid samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Radium-2261
CASRN
13982-63-3
1 The method will measure decay products of these isotopes
This method is for determination of all alpha-emitting radium isotopes by alpha decay analysis. Lead and
barium carriers are added to the sample containing alkaline citrate, then sulfuric acid is added to
precipitate radium, barium, and lead as sulfates. The precipitate is purified by washing with nitric acid,
dissolving in alkaline ethylenediaminetetraacetic acid (EDTA), and re-precipitating as radium-barium
sulfate after pH adjustment to 4.5. This slightly acidic EDTA keeps other naturally occurring alpha-
emitters and the lead carrier in solution. Radium-223, -224, and -226 are identified by the rate of
ingrowth of their daughter products in barium sulfate precipitate. The results are corrected by the rate of
ingrowth to determine radium activity. This method involves alpha counting by a gas-flow internal
proportional counter, scintillation counter, or thin end-window gas-flow proportional counter.
SAM Revision 3.1 103 November 15, 2007
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Section 6 - Selected Radiochemical Methods
Source: Standard Methods for Examination of Water and Wastewater, 21s Edition, American Public
Health Association (APHA), American Water Works Association (AWWA), and Water Environment
Federation (WEF), 2005. Web: http://www.standardmcthods.org/
6.2.24 Standard Method 7500-Ra C: Radium: Emanation Method
This method should be used for confirmatory analysis of aqueous/liquid samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Radium-2261
CASRN
13982-63-3
1 The method will measure decay products of these isotopes
This method is for determination of radium-226 by alpha counting. Radium in water is concentrated and
separated from sample solids by coprecipitation with a relatively large amount of barium as the sulfate.
The precipitate is treated to remove silicates, if present, and to decompose insoluble radium compounds,
fumed with phosphoric acid to remove sulfite, and dissolved in hydrochloric acid. The completely
dissolved radium is placed in a bubbler, which is then closed and stored for a period of several days to 4
weeks for ingrowth of radon. The bubbler is connected to an evacuation system and the radon gas is
removed from the liquid by aeration and helium, dried with a desicant, and collected in a counting cell.
Four hours after radon collection, the cell is counted. The activity of the radon is equal to the radium
concentration. The minimum detectable concentration depends on counter characteristics, background-
counting rate of scintillation cell, cell efficiency, length of counting period, and contamination of
apparatus and environment by radium-226. Without reagent purification, the overall reagent blank
(excluding background) should be between 0.03 and 0.05 pCi radium-226, which may be considered the
minimum detectable amount under routine conditions.
Source: Standard Methods for Examination of Water and Wastewater, 21st Edition, American Public
Health Association (APHA), American Water Works Association (AWWA), and Water Environment
Federation (WEF), 2005. Web: ://www. .org/
6.2.25 Standard Method 7500-Sr B: Total Radioactive Strontium and Strontium-90:
Precipitation Method
This method should be used for qualitative and confirmatory analysis of drinking water and
aqueous/liquid samples for the contaminant identified below and listed in Appendix B.
Analyte(s)
Strontium-901
CASRN
10098-97-2
1 The method will measure decay products of these isotopes
A known amount of inactive strontium ions, in the form of strontium nitrate, is added as a "carrier." The
carrier, alkaline earths, and rare earths are precipitated as the carbonate to concentrate the radiostrontium.
The carrier, along with the radionuclides of strontium, is separated from other radioactive elements and
inactive sample solids by precipitation as strontium nitrate using fuming nitric acid solution. The carrier
and radionuclides of strontium are precipitated as strontium carbonate, which is dried, weighed to
determine recovery of carrier, and measured for radioactivity. The activity of the final precipitate is due
to radioactive strontium only, because all other radioactive elements have been removed. Because it is
impossible to separate the isotopes of strontium-89 and strontium-90 by any chemical procedure, the
amount of strontium-90 is determined by separating and measuring the activity of yttrium-90, its daughter
product. This method involves beta counting by a gas-flow internal proportional counter or thin end-
window low-background proportional counter. A correction is applied to compensate for loss of carriers
and activity during the various purification steps.
SAM Revision 3.1 104 November 15, 2007
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Section 6 - Selected Radiochemical Methods
Source: Standard Methods for Examination of Water and Wastewater, 21s Edition, American Public
Health Association (APHA), American Water Works Association (AWWA), and Water Environment
Federation (WEF), 2005. Web: http://www.standardmcthods.org/
6.2.26 Standard Method 7500-U B: Uranium: Radiochemical Method
This method should be used for qualitative determination in aqueous/liquid samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Uranium-2381
CASRN
7440-61-1
1 If it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
The sample is acidified with hydrochloric or nitric acid and boiled to eliminate carbonate and bicarbonate
ions. Uranium is coprecipitated with ferric hydroxide and subsequently separated. The ferric hydroxide
is dissolved, passed through an anion-exchange column, and washed with acid, and the uranium is eluted
with dilute hydrochloric acid. The acid eluate is evaporated to near dryness, the residual salt is converted
to nitrate, and the alpha activity is counted by a gas-flow proportional counter or alpha scintillation
counter.
Source: Standard Methods for Examination of Water and Wastewater, 21st Edition, American Public
Health Association (APHA), American Water Works Association (AWWA), and Water Environment
Federation (WEF), 2005. Web: http://www.standardmcthods.org/
6.2.27 Standard Method 7500-U C: Uranium: Isotopic Method
This method should be used for confirmatory analysis of aqueous/liquid samples for the contaminant
identified below and listed in Appendix B.
Analyte(s)
Uranium-2381
CASRN
7440-61-1
1 If it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal
digestion methods) or if there is a matrix interference problem, use ORISE Method AP11
This method is a radiochemical procedure for determination of the isotopic content of uranium alpha
activity; it is consistent with determining the differences among naturally occurring, depleted, and
enriched uranium. The sample is acidified with hydrochloric or nitric acid, and uranium-232 is added as
an isotopic tracer. Uranium is coprecipitated with ferric hydroxide and subsequently separated. The
ferric hydroxide is dissolved, passed through an anion-exchange column, and washed with acid, and the
uranium is eluted with dilute hydrochloric acid. The acid eluate is evaporated to near dryness, and the
residual salt is converted to nitrate and electrodeposited onto a stainless steel disc and counted by alpha
spectrometry.
Source: Standard Methods for Examination of Water and Wastewater, 21st Edition, American Public
Health Association (APHA), American Water Works Association (AWWA), and Water Environment
Federation (WEF), 2005. Web:
SAM Revision 3.1 105 November 15, 2007
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Section 6 - Selected Radiochemical Methods
SAM Revision 3.1 106 November 15, 2007
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Section 7 - Selected Pathosen Methods
Section 7.0: Selected Pathogen Methods
A list of the most appropriate methods currently available for use in analyzing environmental samples for
pathogens is provided in Appendix C. This list represents an initial effort towards the goal of providing
standardized analytical procedures. These methods should be used to support remediation activities (site
assessment through clearance) following a homeland security event. The purpose of this section is to
provide summary information regarding the procedures listed. Methods are listed for each pathogen that
may need to be measured and analyzed following an event. Appendix C is sorted alphabetically within
pathogen categories (i.e., bacteria, viruses, protozoa, and helminths).
Protocols from peer-reviewed journal articles are listed for pathogens where standardized methods are not
currently available. Future steps include the development and verification of standardized methods. The
literature references will be replaced as standardized, verified protocols become available.
Pathogens that are categorized as Biosafety Level 4 (BSL-4), such as hemorrhagic fever viruses and
smallpox, will be handled only by reference laboratories with BSL-4 capability and are not included in
this document. Pathogens that are catergorized as Biosafety Level-3 (BSL-3) select agents will be
handled by appropriate Laboratory Response Network (LRN) facilities and are discussed in Section 7.2.1,
with the exception of influenza H5N1 virus and Chlamydophila psittaci, which are analyzed using SAM
procedures. All other pathogens are to be handled as BioSafety Level-2 (BSL-2) using SAM procedures.
Pathogens that are considered to be solely of agricultural concern (i.e., animal and plant pathogens) also
are not currently included. Such pathogens may be considered for possible inclusion in future document
revisions. Some of the pathogens addressed by this document are commonly found in the environment,
and the methods listed in Appendix C assume that analyses will be used to evaluate contamination levels
that are above background conditions. If possible, an investigation of initial background levels, as well as
appropriate controls for background levels, should be performed. It is anticipated that additional site- or
event-specific information also will be required to support the analytical results.
Selection of methods from Appendix C should be based on specific data and information needs, including
whether there is a need to determine the presence of a pathogen, the viability of a pathogen, or both
presence and viability. Although culture-based methods have been selected for many of the pathogens,
molecular techniques such as polymerase chain reaction (PCR) will likely be used for viruses because of
the difficulty and time required to propagate these agents in host cell cultures. It should be noted that,
although molecular techniques are appropriate for evaluation of the presence of a pathogen, these
techniques have inherent limitations with regard to the determination of the viability or infectivity of the
pathogen. In situations where viability determinations are required (e.g., evaluation of the efficacy of
disinfection), viability procedures should be used. Viability procedures are listed for each pathogen
where available.
Appendix C includes the following information:
Pathogen(s). A specific causative agent (e.g., viruses, bacteria) of disease.
• Viability. Ability to grow and/or develop. If the analytical technique can assess viability, it is
denoted by a "yes" in Appendix C. If a technique is not able to assess viability, it is denoted by a
"no."
Analytical technique. An analytical instrument or procedure used to determine the identity,
quantity, and/or viability of a pathogen.
• Analytical method. The unique identifier or number assigned to an analytical method by the method
publisher.
Solid (soil, powder). The recommended method/procedure for the pathogen of interest in solid
samples such as soil and powders.
SAM Revision 3.1 107 November 15, 2007
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Section 7 - Selected Pathosen Methods
• Particulate (swabs, wipes, filters). The recommended method/procedure to measure the pathogen of
interest in participate sample collection devices such as swabs, wipes and high efficiency particulate
air (HEPA) filters.
Liquid/water (filter, grab). The recommended method/procedure for the pathogen of interest in
liquid or aqueous samples that have been collected using a filter or as a grab sample.
• Drinking water (filter, grab). The recommended method/procedure for the pathogen of interest in
potable water samples that have been collected using a filter or as a grab sample.
Aerosol (growth media, filter, liquid). The recommended method/procedure for the pathogen of
interest in air sample collectors such as growth media, filters, or liquid.
Some of the methods in Appendix C include multiple analytical techniques by inference. The analytical
technique listed in Appendix C is intended to be a description of the predominant technique that is
required to provide the data quality parameter (viability or detection and identification). This description
does not preclude the use of other techniques that are within or referenced by the method. For example, a
viability test listed as "culture" may include immunochemical or PCR based assays for the identification
of the culture colony isolates.
Several of the methods listed in Appendix C also include options such as the potential for use of multiple
cell culture media for primary isolation, allowance for selection of a defined subset of a larger number of
biochemical tests for biochemical testing, or use of alternative devices for sample concentration. The
method may provide guidance as to which options should be used under particular circumstances, or this
may be left to the discretion of the laboratory.
7.1 General Guidance
This section provides a general overview of how to identify the appropriate pathogen method(s) for a
given pathogen as well as recommendations for quality control procedures.
For additional information on the properties of the pathogens listed in Appendix C, TOXNET
a cluster of databases on toxicology, hazardous chemicals, and
related areas maintained by the National Library of Medicine, is an excellent resource. Also informative
are CDC's Emergency Preparedness and Response Web site (http://www.bt.cdc.gov/) and the FDA
Center for Food Safety and Applied Nutrition (CFSAN) "Bad Bug Book"
Further research on pathogens is ongoing within EPA, and
databases to manage this information are currently under development.
7.1.1 Standard Operating Procedures for Identifying Pathogen Methods
To determine the appropriate method that is to be used on an environmental sample, locate the pathogen
in Appendix C: Pathogen Methods under the "Pathogen(s)" column. After locating the pathogen,
continue across the table and select an analytical technique that either assesses viability or does not (i.e.,
yes or no, respectively). After an analytical technique has been chosen (e.g., culture, polymerase chain
reaction (PCR), immunoassay), select the analytical method applicable to the sample type of interest
(solid, particulate, liquid/drinking water or aerosol).
Sections 7.2.1 through 7.2.30 below provide summaries of the analytical methods listed in Appendix C.
Once a method has been identified in Appendix C, Table 7-1 can be used to locate the method summary.
SAM Revision 3.1 108 November 15, 2007
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Section 7 - Selected Pathosen Methods
Table 7-1. Pathogen Methods and Corresponding Text Section Numbers
Note: Where procedures are listed in this table for both viability assessment and detection, viability procedures are
listed first.
Pathogen
Method
Section
Bacteria
Campylobacterjejuni
Chlamydophila psittaci
Escherichia coli O157:H7
Leptospira spp.
Usteria monocytogenes
Salmonella spp.
(Method not applicable for
Salmonella typhi)
Salmonella typhi
Shi gel la spp.
Staphylococcus aureus
Vibrio cholerae O1 and O139
SM 9260 G
Molecular and Cellular Probes.
2006. 20: 269-279
Journal of Clinical Microbiology.
2000. 38: 1085-1093
SM 9260 F
SM 9260 I
FDA/Bacteriological Analytical
Manual Chapter 10, 2003
Method 1682
SM 9260 B
SM 9260 E
SM9213B
SM 9260 H
7.2.12
7.2.15
7.2.16
7.2.11
7.2.14
7.2.7
7.2.6
7.2.9
7.2.10
7.2.8
7.2.13
Viruses
Adenoviruses: A-F
Astroviruses
Caliciviruses: Noroviruses
Caliciviruses: Sapoviruses
Coronaviruses: SARS
Hepatitis E Virus
Influenza H5N1 Virus
Picornaviruses: Enteroviruses
Picornaviruses: Hepatitis A Virus
Reoviruses: Rotaviruses
Applied and Environmental
Microbiology. 2005. 71(6): 3131-
3136
Canadian Journal of Microbiology.
2004, 50: 268-278
Journal of Clinical Microbiology.
2004, 42(10): 4679-4658
Journal of Medical Virology, 2006,
78(10): 1347-1353
Journal of Virological Methods.
2004. 122: 29-36
Journal of Virological Methods.
2006. 131(1): 65-71
Emerging Infectious Diseases.
2005. 11(8): 1303-1305
USEPA Manual of Methods for
Virology EPA/600/4-84/013, 2001
Applied and Environmental
Microbiology. 2003. 69(6): 3158-
3164
Applied and Environmental
Microbiology. 2003. 69(6): 3158-
3164
Applied and Environmental
Microbiology. 2003. 69(6): 3158-
3164
7.2.17
7.2.18
7.2.19
7.2.20
7.2.21
7.2.22
7.2.23
7.2.2
7.2.24
7.2.24
7.2.24
Protozoa
Cryptosporidium spp.
Method 1622
Method 1623
Applied and Environmental
Microbiology. 1999. 65(9): 3936-
3941
7.2.4
7.2.5
7.2.25
SAM Revision 3.1
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November 15, 2007
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Section 7 - Selected Pathosen Methods
Entamoeba histolytica
Giardia spp.
Toxoplasma gondii
Journal of Parasitology. 1972.
58(2): 306-310
Journal of Clinical Microbiology.
2005.43(11): 5491-5497
Method 1623
Transactions of the Royal Society of
Tropical Medicine and Hygiene.
1983. 77(4): 487-488
Emerging Infectious Diseases.
2006. 12(2): 326-329
Applied and Environmental
Microbiology. 2004. 70(7): 4035-
4039
7.2.26
7.2.27
7.2.5
7.2.28
7.2.29
7.2.30
Helminths
Baylisascaris procyonis
EPA/625/R92/013
7.2.3
Method summaries are listed in order of method selection hierarchy (see Figure 2-1), starting with EPA
methods followed by methods from other federal agencies, voluntary consensus standard bodies (VCSB),
and journal articles. Methods are listed in numerical order under each publisher. For additional
information regarding sample preparation and analysis procedures available through consensus standards
organizations, please use the contact information provided in Table 7-2.
Table 7-2. Sources of Pathogen Methods
Name
National Environmental Methods Index
(NEMI)
U.S. EPA Microbiology Methods
ICR Microbial Laboratory Manual
USEPA Manual of Methods for Virology
Environmental Regulations and
Technology: Control of Pathogens and
Vector Attraction in Sewage and Sludge
USDA/FSIS Microbiology Laboratory
Guidebook
Bacteriological Analytical Manual
Occupational Safety and Health
Administration Methods
National Institutes for Occupational
Safety and Health Methods
Publisher
U.S. EPA, U.S. Geographical
Society (USGS)
U.S. EPA
U.S. EPA Office of Research and
Development
U.S. EPA
U.S. EPA, National Risk
Management Research
Laboratory (NRMRL)
U.S. Department of Agriculture
(USDA) Food Safety and
Inspection Service
U.S. Food and Drug
Administration (FDA), Center for
Food Safety and Applied Nutrition,
(CFSAN)
Occupational Health and Safety
Administration (OSHA)
National Institute for Occupational
Safety and Health (NIOSH)
Reference
http://vwvw.nemi.qov
http://www.epa.qov/microbes/
http://www.epa.qov/nerlcwww/icrmi
cro.pdf
http://www.epa.qov/nerlcwww/abo
ut.htm
httBL//ww¥j3pjELgc^^
r92013/625r92013.htm
http://www.fsis.usda.aov/Science/
Micj]Dbioio§ic^yj=ab^^^
dj3x,asj3
http://www.dsan. fda.qov/~ebam/b
am~toc.html
http://www.osha.qov
http://www.cdc.qov/niosh/nmam/
SAM Revision 3.1
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November 15, 2007
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Section 7 - Selected Pathosen Methods
Name
Standard Methods for the Examination
of Water and Wastewater, 21st Edition,
2005*
Annual Book of ASTM Standards*
Applied and Environmental
Microbiology*
Journal of Clinical Microbiology*
Clinical Microbiology Procedures
Handbook, 2nd Edition, 2004*
Molecular and Cellular Probes*
Canadian Journal of Microbiology*
Journal of Medical Virology*
Journal of Virological Methods*
Emerging Infectious Diseases
Journal of Parasitology*
Transactions of the Royal Society of
Tropical Medicine and Hygiene*
Diagnostic Procedures in Veterinary
Bacteriology and Mycology
Publisher
American Public Health
Association (APHA), American
Water Works Association
(AWWA), and Water Environment
Federation (WEF)
ASTM International
American Society for Microbiology
American Society for Microbiology
American Society for Microbiology
Elsevier
NRC Research Press
Wiley InterScience
Elsevier
Centers for Disease Control
American Society of
Parasitologists
The Royal Society of Tropical
Medicine and Hygiene
Academic Press
Reference
http://vwvw.standardmethods.orq
httEL/MwvjiMllLoia.
http://aem.asm.orq/
http://icm.asm.orq/
http://estore.asm.orq/viewltemDeta
ils.asp?ltemlD=323
http://www.elsevier.com
Mfe^MMksyiE^S^
http://www3.interscience.wiley.com
/cqi-b in/ho me
MaL//wmj|iswieLt;OTi
http://www.cdc.qov/ncidod/EID/
http://www.bioone.orq
http://www.rstmh.orq/
http://www.pubmedcentraLnih.qov/
' Subscription and/or purchase required.
7.1.2 General Quality Control (QC) Guidance for Pathogen Methods
Generation of analytical data of known and documented quality is a critical factor in the accurate
assessment of and appropriate response to emergency situations. The generation of data of sufficient
quality requires that analytical laboratories: (1) have appropriately trained personnel, (2) acquire and
maintain required supplies, equipment, and reagents, (3) conduct the appropriate QC procedures to ensure
that all measurement systems are in control and operating properly, (4) properly document all analytical
results, and (5) properly document analytical QC procedures and corrective actions.
The level or amount of QC needed depends on the intended purpose of the data generated. Various levels
of QC may be required if the data are generated for presence/absence determinations versus quantitative
results. Specific data needs should be identified, and QC requirements should be based on those needs,
and should be applied consistently across laboratories when multiple laboratories are used. The
individual methods listed, sampling and analytical protocols, or contractual statements of work should be
consulted to determine if additional QC procedures are required.
Method-specific QC requirements are described in many of the methods cited in this manual and will be
included in protocols developed to address specific pathogen/sample type combinations of concern. In
general, analytical QC requirements for pathogen methods include an initial demonstration of
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measurement system capability, as well as the capability of the laboratory and the analyst to perform the
method with the required precision and accuracy.
Ongoing analysis of control samples to ensure the continued reliability of the analytical results should
also be performed. At a minimum, the following QC analyses should be conducted on an ongoing basis:
• Media and reagent sterility checks;
• Positive and negative controls;
Method blanks;
• Reference matrix spikes to evaluate initial and ongoing method/analyst performance, if available;
• Matrix spikes to evaluate method performance in the sample type of interest;
• Matrix spike duplicates (MSB) and/or sample replicates to assess method precision; and
• Instrument calibration checks and temperature controls.
QC procedures should be performed as frequently as necessary to ensure the reliability of analytical
results.
Please note: The appropriate point of contact identified in Section 4 should be consulted regarding
appropriate QA/QC procedures prior to sample analysis. These contacts will consult with the EPA
OSWER coordinator responsible for laboratory activities during the specific event to ensure QA/QC
procedures are performed consistently across laboratories. OSWER is planning to develop QA/QC
guidance for laboratory support. EPA program offices will be responsible for ensuring that the QA/QC
practices are implemented.
7.1.3 Safety and Waste Management
It is imperative that safety precautions be used during collection, processing, and analysis of
environmental samples. Laboratories should have a documented health and safety plan for handling
samples that may contain target chemical, biological, or radiological contaminants, and laboratory staff
should be trained in and implement the safety procedures included in the plan. Pathogens in samples
taken from areas contaminated as the result of a homeland security event may be more hazardous than
naturally occurring pathogens of the same genus and species. The pathogens may have been
manufactured or treated in such a manner as to enhance dispersion or virulence characteristics. These
conditions may warrant special handling for samples arising from intentional contamination incidents. A
laboratory must be made aware of these potential circumstances, and should carefully consider
implementing additional safety measures before agreeing to accept these samples.
In addition, many of the methods listed in Appendix C and summarized or cited in Section 7.2 contain
specific requirements, guidance, or information regarding safety precautions that should be followed
when handling or processing environmental samples and reagents. These methods also provide
information regarding waste management. Other resources that can be consulted for additional
information include the following:
• Environmental Protection Agency's standards regulating hazardous waste (40 CFR parts 260—270),
found at
idx?sid=cac9da30cd241fa70d461c4a917cb75c&c=ccfr&tpl=/ccfrbrowsc/Titlc40/40tab_02.tpl
crobiological and Biomedical Laboratories
Biosafety in Microbiological and Biomedical Laboratories, 4th Edition, found at
"Laboratory Security and Emergency Response Guidance for Laboratories Working with Select
Agents," Morbidity and Mortality Weekly Report, Vol. 5 1, No. RR-19, 1-6, December 6, 2002, found
at http://www.cdc.gov/mmwr/pdf/rr/rr5 1 19.pdf.
Microbiology Biosafety for Level A Laboratories, found at
/A
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OSHA Standards for Hazardous Waste Operations and Emergency Response (29 CFR 1910.120)
found at
http://www.osha.gov/pls/osliawcb/owadisp. show_documcnt?p_tablc=STANDARDS&p_id=9765
OSHA Standards for Occupational Exposure to Hazardous Chemicals in Laboratories (29 CFR
1910.1450) found at
http://www.osha.gov/pls/oshaweb/owadisp. show_document?p_table=STANDARDS&p_id= 10106
OSHA Standards for Respiratory Protection (29 CFR 1910.134) found at
http://w\\rw.osha.gov/pls/osha\veb/o\vadisp.show_document?p_id=12716&p_table=STANDARDS
DOT Hazardous Materials Shipment and Packaging (49 CFR 171-180)
httgiT/ccfi-ggoacccss^goy/cgi/^tcxt/tcxti
idx?sid=585c275ccl9254ba07625d8c92fc925f&c=ccfr&tpl=/ccfrbrowsc/ritlc49/49cfrv2 O2.tpl
Select Agent Rules and Regulations (42 CFR 73 and 9 CFR 121) found at
http://www.acccss.gpo.gov/nara/cfr/waisidx_03/42cfr73_03.html and
http://ccfr.gpoacccss.gov/cgi/t/tcxt/tcxt-idx?c=ccfr&tpl=/ccfrbrowsc/Titlc09/9cfrl21 O2.tpl
7.2 Method Summaries
Summaries of the analytical methods listed in Appendix C are provided in Sections 7.2.1 through 7.2.30.
Each summary contains a table identifying the pathogen(s) and sample type to which the method applies,
a brief description of the method, performance data (if available), and a link to, or source for, obtaining a
full version of the method.
7.2.1 Laboratory Response Network (LRN)
The agents identified below and listed in Appendix C are included in the U.S. Department of Health and
Human Services/U.S. Department of Agriculture (HHS/USDA) select agent list and should be analyzed in
accordance with appropriate LRN protocols.
The LRN was created in accordance with Presidential Directive 39, which established terrorism
preparedness responsibilities for federal agencies. The LRN is primarily a national network of local,
state, federal, military, food, agricultural, veterinary, and environmental laboratories; however, additional
LRN laboratories are located in strategic international locations. The Centers for Disease Control and
Prevention (CDC) provides technical and scientific support to member laboratories as well as secure
access to standardized procedures and reagents for rapid (within 4-6 hours) presumptive detection of
biothreat agents and emerging infectious disease agents. These rapid presumptive assays are part of
sample type/analyte specific algorithms of assays which lead to a confirmed result. The algorithm for a
confirmed result is often a combination of one or more presumptive positive results from a rapid assay in
combination with a positive result from one of the "gold standard" methods, such as culture. The
standardized procedures, reagents, and agent-specific algorithms are considered to be sensitive and are
available only to LRN member laboratories. Thus, these procedures are not available to the general
public and are not discussed in this document.
It is important to note that, in some cases, the procedures may not be fully developed or validated for each
environmental sample type/pathogen combination listed in Appendix C, nor are all LRN member
laboratories necessarily capable of analyzing all of the sample type/pathogen combinations.
Pathogen(s)
Bacillus anthracis [Anthrax]
Brucella spp. [Brucellosis]
Agent Category
Bacteria
Bacteria
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Pathogen(s)
Burkholderia mallei [Glanders]
Burkholderia pseudomallei [Melioidosis]
Coxiella burnetii [Q-fever]
Francisella tularensis [Tularemia]
Yersinia pestis [Plague]
Orthopoxviruses (Variola, Vaccinia, Cowpox,
Monkeypox, Camelpox, Ectromelia, and Gerbilpox)
Agent Category
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Viruses
Please note: Not all methods have been verified for the pathogen/sample type combination listed in
Appendix C. Please refer to the agent-specific method to identify the pathogen/sample type combinations
that have been verified. Any questions regarding information discussed in this section should be referred
to the appropriate contact(s) listed in Section 4.
For additional information on the LRN, including selection of a laboratory capable of receiving and
processing the specified sample type/pathogen, please use the contact information provided below or visit
: //www .bt. cdc. gov/1 ml.
Centers for Disease Control and Prevention
Laboratory Response Branch
Division of Bioterrorism Preparedness and Response (DBPR)
National Center for the Prevention, Detection, and Control of Infectious Diseases (NCPDCID)
Coordinating Center for Infectious Diseases (CCID)
Centers for Disease Control and Prevention (CDC)
1600 Clifton Road NE, Mailstop C-18
Atlanta, GA 30333
Telephone: (404) 639-2790
E-mail: jm;J7lcdc.gov
Local public health laboratories, private laboratories, and commercial laboratories with questions about
the LRN should contact their state public health laboratory director or the Association of Public Health
Laboratories (contact information provided below).
Association of Public Health Laboratories
8515 Georgia Avenue, Suite 700
Silver Spring, MD 20910 Telephone: (240) 485-2745
Fax: (240) 485-2700
Web site: www.aphl.org
E-mail: info@apM.org
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7.2.2 USEPA Manual of Methods for Virology, EPA/600/4-84/013, April 2001
This method should be used for detection and viability assessment of picornaviruses (enteroviruses) in
solid, particulate, liquid, and water samples. Note: This manual also describes procedures for preparation
of samples for non-LRN viruses [i.e., adenovirus, astrovirus, norovirus, sapovirus, coronavirus (SARS),
hepatitis E virus, influenza H5N1 virus, picornaviruses (enterovirus and hepatitis A virus), and reovirus
(rotavirus)].
Pathogen(s)
Picornaviruses: Enteroviruses
Agent Category
Viruses
This manual describes procedures for determining the infectivity of enteroviruses, including a
neutralization test used to identify these viruses. The test uses reference-typing sera directed against
isolated waterborne viruses, and consists of simultaneously inoculating virus and antiserum into a
microtiter plate, incubating the virus-antibody mixture for two hours, adding a suspension of host cells to
the mixture, incubating the host cells-virus-antibody mixture for three days, and then examining the cells
daily for five more days for the absence or presence of cytopathic effect (CPE). The test uses the Lim
Benyesh-Melnick (LB-M) antiserum pools, which consist of 61 equine antisera, including LB-M
antiserum pools A-H for the identification of 41 enteroviruses. Chapters 7 and 14 in this manual describe
procedures for the collection and preparation of virus samples. Sample preparation procedures described
include concentration and processing of waterborne viruses by positively charged 1MDS cartridge filters
and flocculation. These general procedures can be used for many viruses and may be adapted for analysis
of particulate, liquid, water and aerosol samples. (See Footnote 4 of Appendix C.)
Please note: These procedures have been applied to analysis of water samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: USEPA Manual of Methods for Virology EPA/600/4-84/013. April 2001. Identification of
Enteroviruses. Chapter 12. (http://www.epa.gov/microbes/chaptl2.pdf). For information regarding this
manual, please contact Ann Grimm (see Section 4 for contact information).
7.2.3 USEPA Environmental Regulations and Technology, Control of Pathogens and
Vector Attraction in Sewage Sludge EPA/625/R-92/013, July 2003: Baylisascaris
procyonis
These protocols should be used for detection and viability assessment of Baylisascaris procyonis in
solid, particulate, liquid, and water samples.
Pathogen(s)
Baylisascaris procyonis
Agent Category
Helminths
The protocol describes procedures for analysis of solid and wastewater samples and may be adapted to
analysis of particulate, liquid, water, and aerosol samples. Samples are processed by blending with
buffered water containing a surfactant. The blend is screened to remove large particles, the solids in the
screened portion are allowed to settle out, and the supernatant is decanted. The sediment is subjected to
density gradient centrifugation using magnesium sulfate. This flotation procedure yields a layer likely to
contain Ascaris and other parasite ova, if present in the sample. Small particulates are removed by a
second screening on a small mesh size screen. The resulting concentrate is incubated until control
Ascaris eggs are fully embryonated. The concentrate is then microscopically examined for the categories
of Ascaris ova on a counting chamber.
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Please note: This protocol has been applied to analysis of wastewater, sludge, and compost samples.
Further research is needed to develop and standardize the protocols for other sample types. At a
minimum, the following quality control checks should be performed and evaluated before using this
protocol: positive control, negative control, and blank.
Source: USEPA Environmental Regulations and Technology: Control of Pathogens and Vector
Attractions in Sewage Sludge, "Appendix I: Test Method for Detecting, Enumerating, and Determining
the Viability of Ascaris Ova in Sludge." EPA/625/R-92/013. July 2003.
7.2.4 EPA Method 1622: Cryptosporidium in Water by Filtration/IMS/FA
This method should be used for detection of Cryptosporidium spp. in solid, particulate, and drinking
water samples, but is not suitable for determining viability.
Pathogen(s)
Cryptosporidium spp. [Cryptosporidiosis]
Agent Category
Protozoa
This method describes procedures for analysis of drinking water samples and may be adapted for analysis
of solid and particulate samples. A water sample is filtered and the oocysts and extraneous materials are
retained on the filter. Materials on the filter are eluted, the eluate is centrifuged to pellet the oocysts, and
the supernatant fluid is aspirated. The oocysts are magnetized by attachment of magnetic beads
conjugated to anti-Cryptosporidium antibodies. The magnetized oocysts are separated from the
extraneous materials using a magnet, and the extraneous materials are discarded. The magnetic bead
complex is then detached from the oocysts. The oocysts are stained on well slides with fluorescently
labeled monoclonal antibodies and 4',6-diamidino-2-phenylindole (DAPI). The stained sample is
examined using fluorescence and differential interference contrast (DIC) microscopy. Qualitative
analysis is performed by scanning each slide well for objects that meet the size, shape, and fluorescence
characteristics of Cryptosporidium oocysts. Quantitative analysis is performed by counting the total
number of objects on the slide confirmed as oocysts. This method is not intended to determine viability
of the oocysts.
Please note: This method has been applied to analysis of drinking water samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, matrix spike and matrix spike duplicate (MS/MSD), and blank.
Source: USEPA. 2005. Cryptosporidium in Water by Filtration/IMS/FA. United States Environmental
Protection Agency, Washington, D.C. (http://www.cpa.gov/ncrlcwww/1622dc05.pdf)
7.2.5 EPA Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA
This method should be used for the detection of Cryptosporidium spp. in solid, particulate, and
liquid/water samples and Giardia spp. in solid, particulate, and liquid/water samples, but is not suitable
for determining viability.
Pathogen(s)
Cryptosporidium spp.
Giardia spp.
Agent Category
Protozoa
Protozoa
This method describes procedures for analysis of wastewater samples and may be adapted for assessment
of solid, particulate, and liquid samples. A water sample is filtered and the oocysts and cysts and
extraneous materials are retained on the filter. Materials on the filter are eluted, the eluate is centrifuged
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to pellet the oocysts and cysts, and the supernatant fluid is aspirated. The oocysts and cysts are
magnetized by attachment of magnetic beads conjugated to wii-Cryptosporidium and anii-Giardia
antibodies. The magnetized oocysts and cysts are separated from the extraneous materials using a
magnet, and the extraneous materials are discarded. The magnetic bead complex is then detached from
the oocysts and cysts. The oocysts and cysts are stained on well slides with fluorescently labeled
monoclonal antibodies and 4',6-diamidino-2-phenylindole (DAPI). The stained sample is examined using
fluorescence and differential interference contrast (DIC) microscopy. Qualitative analysis is performed
by scanning each slide well for objects that meet the size, shape, and fluorescence characteristics of
Cryptosporidium oocysts and Giardia cysts. Quantitative analysis is performed by counting the total
number of objects on the slide confirmed as oocysts or cysts. This method is not intended to determine
viability of the parasites.
Please note: This method has been applied to analysis of wastewater samples. Further research is needed
to develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, matrix spike and matrix spike duplicate (MS/MSD), and blank.
Source: USEPA. 2001. Cryptosporidium and Giardia in Water by Filtration/IMS/FA. United States
Environmental Protection Agency, Washington, B.C. (http://www.cpa.gov/microbcs/1623dc05.pdf)
7.2.6 EPA Method 1682: Salmonella spp.
This method should be used for detection and viability assessment ofSamonella spp. (except S. typhi) in
solid, particulate, and liquid/water samples.
Pathogen(s)
Salmonella spp.
(Method not applicable to Salmonella typhi)
Agent Category
Bacteria
This method describes procedures for analysis of solid samples (biosolids) and may be adapted for
assessment of water, liquid, particulate, and aerosol samples. Prepared samples are inoculated into tubes
of tryptic soy broth (TSB) and incubated for 24 hours. Positive (turbid) tubes are spotted onto plates of
modified semisolid Rappaport-Vassiliadis (MSRV) medium and incubated at 42°C for 16 to 18 hours.
The MSRV medium uses novobiocin and malachite green to inhibit non-Salmonella species, while
allowing most Salmonella species to grow. MSRV and the elevated incubation temperature (42°C) are
inhibitory for S. typhi. Presumptive colonies are isolated on xylose-lysine desoxycholate (XLD) agar and
confirmed using lysine-iron (LIA), triple sugar iron (TSI) agar, and urea broth, followed by serological
typing using polyvalent O antisera.
Please note: This procedure has been applied to analysis of solid (biosolids) samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: USEPA. 2006. Salmonella in Sewage Sludge (Biosolids) by Modified Semisolid Rappaport-
Vassiliadis (MSRV) Medium. United States Environmental Protection Agency, Washington, D.C.
(http://www.epa.gov/waterscience/methods/method/biological/1682.pdf)
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7.2.7 U.S. FDA Bacteriological Analytical Manual, Chapter 10, 2003: Listeria
monocytogenes
This method should be used for detection and viability assessment of Listeria monocytogenes in solid,
particulate, aerosol, liquid and water samples.
Pathogen(s)
Listeria monocytogenes
Agent Category
Bacteria
Procedures are described for analysis of food samples and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. Prepared samples are incubated for 4 hours in Buffered
Listeria Enrichment Broth (BLEB) without selective agents. Cycloheximide is added and incubation
continued. At 24 and 48 hours, BLEB cultures are streaked onto esculin-containing selective isolation
agar (i.e., Oxford Medium, OXA) and incubated for an additional 24 to 48 hours. A L. monocytogenes-L.
ivanovii differential selective agar, such as Biosynth Chromogen Medium (BCM), is streaked at 48 hours.
Presumptive Listeria colonies are black with a black halo on esculin-containing media and blue on BCM
plates. Isolated colonies are streaked onto Trypticase soy agar with yeast extract (TSAye), incubated for
24 to 48 hours, and examined for morphological and biochemical characteristics. L. monocytogenes is a
rod-shaped Gram-positive, motile bacterium. It is catalase positive, ramnose-positive, and mannitol- and
xylose-negative. Purified isolates may be rapidly identified using commercially available biochemical
typing kits. Confirmation is performed with commercially available sera.
Please note: This method has been applied to analysis of food samples. Further research is needed to
develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, and blank.
Source: U.S. Food & Drug Administration, Center for Food Safety & Applied Nutrition. 2001.
Bacteriological Analytical Manual Online. Chapter 10. (http://www.cfsan.fda.gov/~cbam/bam-16.html)
7.2.8 Standard Methods 9213 B: Staphylococcus aureus
This method should be used for detection and viability assessment of Staphylococcus aureus in solid,
particulate, aerosol, liquid, and water samples.
Pathogen(s)
Staphylococcus aureus
Agent Category
Bacteria
Procedures are described for analysis of water samples and may be adapted for assessment of solid,
liquid, particulate, and aerosol samples. Prepared samples are inoculated into tubes of M-staphylococcus
broth and incubated for 24 hours. Positive (turbid) tubes are streaked onto plates of Baird-Parker agar and
incubated for 48 hours. Presumptive S. aureus colonies are tested for mannitol fermentation by the
addition of a drop of bromthymol blue, a pH indicator. Isolated colonies are examined for morphological
and biochemical characteristics. S. aureus is a Gram-positive coccus. Biochemical characterizations
include catalase-positive, coagulase-positive, fermentation of mannitol, and anerobic fermentation of
glucose.
Please note: This method has been applied to analysis of water samples. Further research is needed to
develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, and blank.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition. Qittrr//www_.standardjncthods1or^/)
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7.2.9 Standard Methods 9260 B: General Qualitative Isolation and Identification
Procedures for Salmonella
This method should be used for detection and viability assessment of Salmonella typhi in solid,
particulate, liquid, and water samples.
Pathogen(s)
Salmonella typhi [Typhoid fever]
Agent Category
Bacteria
This method describes procedures for analysis of water samples and may be adapted for assessment of
solid, particulate, and liquid samples. Concentrated samples are enriched in either selenite cystine,
selenite-F, or tetrathionate broths and incubated at 35°C to 37°C for up to 5 days. An aliquot from each
turbid tube is streaked onto bismuth sulfite (BS) plates and incubated at 35 °C to 37 °C for 24 to 48 hours.
Presumptive positive colonies are then subjected to biochemical characterization. Confirmation is
through serological testing using polyvalent O and Vi antiserum.
Please note: This method has been applied to analysis of water samples. Further research is needed to
develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, and blank.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition. Qittg^/wwwjtandardjncthods^or^/)
7.2.10 Standard Methods 9260 E: Shigella
This method should be used for detection and viability assessment of Shigella species in solid,
particulate, liquid, and water samples.
Pathogen(s)
Shigella spp. [Shigellosis]
Agent Category
Bacteria
This method describes procedures for analysis of water samples and may be adapted for assessment of
solid, particulate, and liquid samples. This method contains two options for sample concentration:
membrane filtration (liquid samples) and centrifugation (liquid and solid samples) for analyses. Both
options include inoculation of an enrichment medium (Selenite F broth). Isolation of the target pathogen
is achieved by plating onto Xylose Lysine Deoxycholate (XLD) and/or MacConkey agar. Biochemical
identification consists of inoculating Triple Sugar Iron (TSI) and Lysine Iron Agar (LIA) slants.
Confirmation is performed by slide agglutination tests using polyvalent antisera.
Please note: This method has been applied to analysis of water and solid samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition. Qittrr//www1standardjncthods1or^/)
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7.2.11 Standard Methods 9260 F: Pathogenic Escherichia coli
This method should be used for detection and viability assessment of Escherichia coli O157:H7 in solid,
particulate, liquid, and water samples.
Pathogen(s)
Escherichia coli (E. coli) O157:H7
Agent Category
Bacteria
This method describes procedures for analysis of water samples and may be adapted for assessment of
solid, particulate, and liquid samples. The method allows for two options, one being a modification of
SM 9221B followed by plating and biochemical identification. The second option, modification of a food
method, allows for the analysis of large sample volumes. A 200-mL water sample is centrifuged,
resuspended in E. coli enrichment broth (EEB) and incubated for 6 hours. Tellurite Cefixime Sorbitol-
MacConkey (TC SMAC) plates are inoculated with the enriched EEB culture, and incubated for up to 24
hours. Colorless colonies on TC SMAC are tested for indole production. Presumptive positive colonies
are then subjected to biochemical characterization. Confirmation is through serological testing.
Please note: This method has been applied to analysis of water samples. Further research is needed to
develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, and blank.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition, (http://www.staiidardmcthods.org/)
7.2.12 Standard Methods 9260 G: Campylobacterjejuni
This method should be used for detection and viability assessment of Campylobacterjejuni in solid,
particulate, liquid, and water samples.
Pathogen(s)
Campylobacterjejuni
Agent Category
Bacteria
This method describes procedures for analysis of water samples and may be adapted for assessment of
solid, particulate, and liquid samples. Water samples (1 to several liter volumes) are filtered using a
cellulose nitrate membrane filter. Filters are placed face down on Skirrow's medium and incubated for 24
hours at 42° C in a microaerobic environment. Alternatively, samples are enriched in Campylobacter
broth supplemented with antibiotics and lysed horse blood in a microaerobic environment at 37°C for 4
hours, then at 42°C for 24 to 48 hours prior to streaking on Skirrow's medium. Identification is made by
culture examination, microscopy, motility testing, and biochemical testing. Biochemical tests include
oxidase, catalase, nitrite and nitrate reduction, H2S production, and hippurate hydrolysis. Confirmation is
performed using commercially available rapid serological test kits. Skirrow's and other selective media
containing antibiotics (trimethoprim, vancomycin, polymixin) may prevent the growth of injured
organisms.
Please note: This method has been applied to analysis of water samples. Further research is needed to
develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, and blank.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition, (http://www.staiidardmcthods.org/)
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7.2.13 Standard Methods 9260 H: Vibrio cholerae
This method should be used for detection and viability assessment of Vibrio cholerae in solid,
particulate, liquid, and water samples.
Pathogen(s)
Vibrio cholerae [Cholera]
Agent Category
Bacteria
This method describes procedures for analysis of water samples and may be adapted for assessment of
solid, particulate, and liquid samples. Samples are enriched in alkaline peptone broth and incubated for
up to 8 hours. Thiosulfate-citrate-bile salts-sucrose (TCBS) agar plates are inoculated with the incubated
broth and incubated for 24 hours. Yellow sucrose-fermenting colonies are presumptive for V. cholera and
are plated on tryptic soy agar with 0.5% NaCl. Presumptive positive colonies are then subjected to
biochemical characterization. Confirmation is performed using slide agglutination assays for serological
identification.
Please note: This method has been applied to analysis of water samples. Further research is needed to
develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, and blank.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition. (jittgy/www.stgodardmethods±oig/)
7.2.14 Standard Methods 9260 I: Leptospira
This method should be used for detection and viability assessment of Leptospira interrogans in solid,
particulate, liquid, and water samples.
Pathogen(s)
Leptospira interrogans [Leptospirosis]
Agent Category
Bacteria
This method describes procedures for analysis of water samples and may be adapted for assessment of
solid, particulate, and liquid samples. Filter samples through a 0.22-um filter, retaining the filtrate as
inoculum. If the sample is turbid, a succession of filters of decreasing pore size may be used prior to the
0.22-um filter. A tube of Leptospira Medium Base (Ellinghausen-McCullough Johnson Harris
formulation (EMJH), supplemented with bovine serum albumin (BSA) and Tween®), is inoculated with
the sample and incubated at 30°C for up to 6 weeks. Alternatively, the sample is added directly to the
media, incubated overnight, passed through a 0.22-um membrane filter, and incubation continued for up
to 6 weeks. Cultures are examined by darkfield microscopy for motile leptospires. Confirmation is
performed by microscopic agglutination test using reference antisera.
Please note: This method has been applied to analysis of water samples. Further research is needed to
develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, and blank.
Source: American Public Health Association, American Waterworks Association, and Water
Environment Federation. 2005. Standard Methods for the Examination of Water and Wastewater. 21st
Edition, (http://www.standardmethods.org/)
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7.2.15 Literature Reference for Campylobacterjejuni (Molecular and Cellular Probes.
2006. 20: 269-279)
These procedures should be used for detection of Campylobacterjejuni in solid, particulate, aerosol,
liquid, and water samples, but are not suitable for determining viability.
Pathogen(s)
Campylobacterjejuni
Agent Category
Bacteria
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. The method uses real-time quantitative polymerase chain
reaction (PCR) for identification of C. jejuni. A high through-put method using guanidinium thiocyanate
(GTC) and glass beads is used for extraction of deoxyribonucleic acid (DNA). C. jejuni uses the CJ F
primer, CJ R primer, and CJ probe for real-time quantitative PCR. Results are evaluated against standard
curves made with a 10-fold dilution series of C. jejuni NCTC 11168 DNA in a background of cecum
DNA.
Please note: These procedures have been applied to analysis of clinical samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. PCR quality control checks should be performed according to EPA Draft
Quality Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on
Environmental Samples document (wj^¥X)i3a4?OT/nc^^ or consult the point of
contact identified in Section 4.
Source: Skanseng, B., Kaldhusdal, M., and Rudi, K. 2006. "Comparison of Chicken Gut Colonization by
the Pathogens Campylobacterjejuni and Clostridium perfringens by the Real-time Quantitative PCR."
Molecular and Cellular Probes. 20: 269-279.
7.2.16 Literature Reference for Chlamydophila psittaci (Journal of Clinical Microbiology.
2000.38: 1085-1093)
These procedures should be used for detection and viability assessment of Chlamydophila psittaci in
solid, particulate, aerosol, liquid and water samples.
Pathogen(s)
Chlamydophila psittaci
(formerly known as Chlamydia psittaci)
Agent Category
Bacteria
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. The method uses touchdown enzyme time release-
polymerase chain reaction (TETR-PCR) for detection and identification of Chlamydophila psittaci.
Deoxyribonucleic acid (DNA) is extracted from chlamydia cultures (Buffalo Green Monkey Kidney
(BGMK) cells), minced clinical tissues, and respiratory samples by mixing with a chelating resin, heating
first to 56°C for 15 to 30 minutes, then 100°C for 8 to 10 minutes. Primer sets specific for C. psittaci are
designed based on the DNA sequences of the 16S ribosomal ribonucleic acid (rRNA) and 16S-23S rRNA
genes (CPS 100/101). PCR products are separated by electrophoresis in 12% polyacrylamide gels with
Tris-borate-ethylenediaminetetraacetic acid (EDTA) buffer and visualized with ethidium bromide.
Chlamydophila psittaci samples are to be assessed at BioSafety Level-3 (BSL-3) laboratories.
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Please note: These procedures have been applied to analysis of clinical (vaginal and respiratory) samples.
Further research is needed to develop and standardize the protocols for other sample types. At a
minimum, the following quality control checks should be performed and evaluated before using this
protocol: positive control, negative control, and blank. PCR quality control checks should be performed
according to EPA Draft Quality Assurance/Quality Control Guidance for Laboratories Performing PCR
Analyses on Environmental Samples document (www.cpa.gov/ncrlcwww/qa_qc_pciiO_04.pdf) or consult
the point of contact identified in Section 4.
Source: Madico, G., Quinn, T.C., Boman, J., and Gaydos, C.A. 2000. "Touchdown Enzyme Time
Release-PCR for Detection and Identification ofChlamydia trachomatis, C. pneumoniae, and C. psittaci
Using the 16S and 16S-23S Spacer rRNA Genes." Journal of Clinical Microbiology. 38(3): 1085-1093.
j3ub^^
7.2.17 Literature Reference for Adenoviruses (Applied and Environmental Microbiology.
2005.71(6): 3131-3136)
These procedures should be used for detection and viability assessment of Adenoviruses, enteric and
non-enteric (A-F) in solid, particulate, aerosol, liquid, and water samples. Note: Procedures described in
the USEPA Manual of Methods for Virology (EPA/600/4-84/013, April 2001) can be adapted for the
)reparation of samples.
Pathogen(s)
Adenoviruses: Subgroups A-F
Agent Category
Viruses
Procedures are described for analysis of cell culture lysates and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. The detection procedure uses a broadly reactive
fluorogenic 5' nuclease (TaqMan®) quantitative real-time polymerase chain reaction (PCR) assay for the
detection of all six species (A-F) of human adenoviruses (HAdV). Sensitive detection and discrimination
of adenovirus F species (AdV40 and AdV41) can be achieved by using a real-time fluorescence
resonance energy transfer (FRET)-based PCR assay.
For the viability assessment of adenovirus 40 and 41, given that they can be difficult to grow in culture,
cell lines such as G293 (Journal of Medical Virology, 11(3): 215-231) or CaCo-2 (Journal of Medical
Virology. 1994. 44(3): 310-315) may be considered when these viruses are suspected to be present. As
detection of adenovirus in environmental samples can be difficult, additional methods such as described
in Effect of Adenovirus Resistance on UV Disinfection Experiments: A Report on the State of Adenovirus
Science (J. AWWA. 2006. 98(6):93-106) also may be useful.
Please note: These procedures have been applied to analysis of cell culture lysates. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. PCR quality control checks should be performed according to EPA Draft
Quality Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on
Environmental Samples document (w^w.ega^ggv/oerjcw^w/ga_gc_gcd0_04_±pdf) or consult the point of
contact identified in Section 4.
Source: Jothikumar, N., Cromeans, T. L., Hill, V. R., Lu, X., Sobsey, M., and Erdman, D. D. 2005.
"Quantitative Real-Time PCR Assays for Detection of Human Adenoviruses and Identification of
Serotypes 40 and 41." Applied and Environmental Microbiology. 71 (6): 3131-3136.
http://aem.asm.Org/cgi/reprint/71/6/3131
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7.2.18 Literature Reference for Astroviruses (Canadian Journal of Microbiology. 2004. 50:
269-278)
These procedures should be used for detection and viability assessment of Astroviruses in solid,
particulate, aerosol, liquid, and water samples. Note: Procedures described in the USEPA Manual of
Methods for Virology (EPA/600/4-84/013, April 2001) can be adapted for the preparation of samples.
Pathogen(s)
Astroviruses
Agent Category
Viruses
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. The method detects eight astrovirus serotypes. The
method is a reverse transcription-polymerase chain reaction (RT-PCR) procedure optimized for use in a
real-time PCR assay and can be integrated with sample-cell culture (CaCo-2 cells) to enhance sensitivity
(ICC/RT-PCR). Water samples are collected by filtration (1MDS filter), and viruses are eluted using a
beef extract solution (1.5%, pH 9.5). Viruses are concentrated using celite adsorption (pH 4.0), filtration,
and celite-elution with sodium phosphate (0.15 M, pH 9.0), followed by further concentration and
processing to remove inhibitors (ultracentrifugation, solvent extraction, and MW-exclusion filtration).
Concentrated samples are analyzed directly or indirectly (following cell culture) by a two-step RT-PCR
(RT followed by PCR) assay using astrovirus-specific primer sets. Detection of amplicons is by gel
electrophoresis with subsequent confirmation by hybridization (dot-blot) using digoxigenin-labeled
internal (nested) probes or by real-time detection using fluorogenic probes.
Please note: These procedures have been applied to analysis of clinical (fecal) samples. Further research
is needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. PCR quality control checks should be performed according to EPA Draft
Quality Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on
Environmental Samples document (www.cpa.gov/ncrlcwww/qa qc per 10 04.pdf) or consult the point of
contact identified in Section 4.
Source: Grimm, A. C., Cashdollar, J. L., Williams, F. P., and Fout, G. S. 2004. "Development of an
Astrovirus RT-PCR Detection Assay for Use with Conventional, Real-Time, and Integrated Cell
Culture/RT-PCR." Canadian Journal of Microbiology. 50(4): 269-268.
7.2.19 Literature Reference for Noroviruses (Journal of Clinical Microbiology. 2004.
42(10): 4679-4685)
These procedures should be used for the detection assessment of noroviruses in solid, particulate, aerosol,
liquid, and water samples, but are not suitable for determining viability. Note: Procedures described in
the USEPA Manual of Methods for Virology (EPA/600/4-84/013, April 2001) can be adapted for the
)reparation of samples.
Pathogen(s)
Caliciviruses: Noroviruses
Agent Category
Viruses
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. This method is an assay for the detection and quantitation
of norovirus using LightCycler® real-time reverse transcription-polymerase chain reaction (real-time LC
RT-PCR) technology. Viral ribonucleic acid (RNA) is extracted using either a commercial kit or a silica-
based method. For Norovirus G-l, primers based on the capsid gene sequence are used, and for
Norovirus G-II, primers based on the polymerase gene sequence are used. A SYBR® Green I system is
used in the reaction for visualization. External standard curves for the quantification of norovirus are
established using RNA transcripts from strains S5 and S19, corresponding to G-I/4 and G-II/12,
respectively.
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Please note: These procedures have been applied to analysis of clinical (fecal) samples. Further research
is needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. PCR quality control checks should be performed according to EPA Draft
Quality Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on
Environmental Samples document (www.cpa.gov/ncrlcwww/qa qc per 10 04.pdf) or consult the point of
contact identified in Section 4.
Source: Pang, X., Lee, B., Chui, L., Preiksaitis, J.K., and Monroe, S.S. 2004. "Evaluation and Validation
of Real-Time Reverse Transcription-PCR Assay Using the LightCycler System for Detection and
Quantitation of Noro virus. "Journal of Clinical Microbiology. 42(10): 4679-4685.
http: III cm. . org/cgi/rcprint/42/10/4679
7.2.20 Literature Reference for Sapoviruses (Journal of Medical Virology. 2006. 78(10):
1347-1353)
These procedures should be used for detection of sapoviruses in solid, particulate, aerosol, liquid, and
water samples, but are not suitable for determining viability. Note: Procedures described in the USEPA
Manual of Methods for Virology (EPA/600/4-84/013, April 2001) can be adapted for the preparation of
samples.
Pathogen(s)
Caliciviruses: Sapoviruses
Agent Category
Viruses
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. The method is a TaqMan®-based real-time reverse
transcriptase polymerase chain reaction (RT-PCR) assay that has the ability to detect four of the five
distinct sapovirus (SaV) genogroups (Gl-GV). Sets of primers, based on the multiple alignment of 27
gene sequences for the polymerase-capsid junction in open reading frame 1 (ORF1), are used to detect
human SaV GI, Gil, GIV, and GV sequences in a single tube. Sensitivity using control plasmids range
from 2.5 X 101 to 2.5 X 107 copies per tube. No cross-reactivity is observed against other enteric viruses,
including norovirus (NoV), rotavirus, astrovirus, and adenovirus.
Please note: These procedures have been applied to analysis of clinical (fecal) samples. Further research
is needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. PCR quality control checks should be performed according to EPA Draft
Quality Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on
Environmental Samples document (w^w.ega^gv/oerJcw^w/gaL_gc_gcdO_04_±pdf) or consult the point of
contact identified in Section 4.
Source: Oka T, Katayama K, Hansman GS, Kageyama T, Ogawa S, Wu FT, White PA, and TakedaN.
2006. "Detection of Human Sapovirus by Real-time Reverse Transcription-Polymerase Chain Reaction."
Journal of Medical Virology. 78(10): 1347-1353.
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7.2.21 Literature Reference for Coronaviruses (SARS) (Journal of Virological Methods.
2004. 122:29-36)
These procedures should be used for detection of severe acute respiratory syndrome-associated human
coronavirus (SARS-HCoV) in solid, particulate, aerosol, liquid, and water samples, but are not suitable
for determining viability. Note: Procedures described in the USEPA Manual of Methods for Virology
(EPA/600/4-84/013, April 2001) can be adapted for the preparation of samples.
Pathogen(s)
Coronaviruses: SARS-associated human coronavirus
Agent Category
Viruses
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. This method uses a conventional single-tube reverse
transcription-polymerase chain reaction (RT-PCR) procedure based on consensus primer sequences
targeting conserved regions of coronavirus genome sequences. End-point amplicon analysis is by
electrophoresis and subsequent visualization. The assay can detect the SARS-HCoV as well as several
other human respiratory Coronaviruses (HCo-OC43 and HCoV-229E). Species identification is provided
by sequencing the amplicon, although rapid screening can be performed by restriction enzyme analysis.
Please note: These procedures have been applied to analysis of clinical (throat, lung, blood, stool, urine)
samples. Further research is needed to develop and standardize the protocols for other sample types. At a
minimum, the following quality control checks should be performed and evaluated before using this
protocol: positive control, negative control, and blank. PCR quality control checks should be performed
according to EPA Draft Quality Assurance/Quality Control Guidance for Laboratories Performing PCR
Analyses on Environmental Samples document (www.cpa.gov/ncrlcwww/qa qc pcrlO 04.pdf) or consult
the point of contact identified in Section 4.
Source: Adachi, D., Johnson. G., Draker, R, Ayers, M., Mazzulli, T., Talbot, P. J., and Tellier, R. 2004.
"Comprehensive Detection and Identification of Human Coronaviruses, Including the SARS-associated
Coronavirus, with a Single RT-PCR Assay." Journal of Virological Methods. 122: 29—36.
7.2.22 Literature Reference for Hepatitis E Virus (Journal of Virological Methods. 2006.
131(1): 65-71)
These procedures should be used for the detection of Hepatitis E virus in solid, particulate, aerosol,
liquid, and water samples, but are not suitable for determining viability. Note: Procedures described in
the USEPA Manual of Methods for Virology (EPA/600/4-84/013, April 2001) can be adapted for the
)reparation of samples.
Pathogen(s)
Hepatitis E virus (HEV)
Agent Category
Viruses
Procedures are described for analysis of spiked water samples and may be adapted for assessment of
solid, particulate, aerosol, and liquid samples. The method uses a TaqMan® real-time reverse
transcriptase-polymerase chain reaction (RT-PCR) assay in order to detect and quantitate all four major
HEV strains that may be present in clinical and environmental samples. Primers and probes are based on
the multiple sequence alignments of 27 gene sequences for the ORF3 region. Thirteen HEV isolates
representing genotypes 1—4 are used to standardize the real-time RT-PCR assay. The assay can detect as
few as four genome equivalent (GE) copies of HEV plasmid DNA and 1.2 50% pig infectious dose
(PID50) of swine HEV. Concentrations of swine HEV from 1.2 to 120 PID50 spiked into a surface water
sample can also be detected.
Please note: These procedures have been applied to analysis of spiked water. Further research is needed
to develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
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control, and blank. PCR quality control checks should be performed according to EPA Draft Quality
Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on Environmental
Samples document (www.cpa.gov/ncrlcwww/qa qc pcrlO 04.pdf) or consult the point of contact
identified in Section 4.
Source: Jothikumar, N., Cromeans, T.L., Robertson, B.H., Meng, X.J., and Hill, V.R. 2006. "A Broadly
Reactive One-step Real-time RT-PCR Assay for Rapid and Sensitive Detection of Hepatitis E Virus."
Journal of VirologicalMethods, Vol. 131(1): 65-71.
7.2.23 Literature Reference for Influenza H5N1 (Emerging Infectious Diseases. 2005.
11(8): 1303-1305)
These procedures should be used for the detection of Influenza H5N1 virus in solid, particulate, aerosol,
liquid, and water samples, but are not suitable for determining viability. Note: Procedures described in
the USEPA Manual of Methods for Virology (EPA/600/4-84/013, April 2001) can be adapted for the
)reparation of samples.
Pathogen(s)
Influenza H5N1 virus
Agent Category
Viruses
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, aerosol, liquid, and water samples. This is a two-step, real-time reverse transcriptase-
polymerase chain reaction (RT-PCR) multiplex assay. It employs a mixture of two sets of primers and
dual-labeled fluorescent probes that specifically target two different regions of the HA gene of H5N1.
Viral ribonucleic acid (RNA) is extracted using a commercial viral RNA extraction kit and reverse
transcribed with random hexamers. Five microliters of the complementary deoxyribonucleic acid
(cDNA) is used for PCR. At the end of each annealing step, the fluorescent signal of each reaction is
measured at a wavelength of 530 nm with the fluorimeter. The assay is specific for the H5 subtype.
Influenza H5N1 virus samples are to be handled at BioSafety Level-3 (BSL-3) laboratories.
Please note: These procedures have been applied to analysis of clinical (respiratory) samples. Further
research is needed to develop and standardize the protocols for other sample types. At a minimum, the
following quality control checks should be performed and evaluated before using this protocol: positive
control, negative control, and blank. PCR quality control checks should be performed according to EPA
Draft Quality Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on
Environmental Samples document (www.cpa.gov/ncrlcwww/qa qc per 10 04.pdf) or consult the point of
contact identified in Section 4.
Source: Ng, E.K.O., Cheng, P.K.C., Ng, A.Y.Y., Hoang, T.L., and Lim, W.W.L. 2005.
"Influenza A H5N1 Detection." Emerging Infectious Diseases. 11(8): 1303-1305.
http://www.cdc.gov/ncidod/EID/vol 1 Ino08/pdfs/04-1317.pdf
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7.2.24 Literature Reference for Enteric Viruses (Applied and Environmental Microbiology.
2003.69(6): 3158-3164)
These procedures should be used for detection of enterovirus and hepatitis A virus (HAV) in solid,
particulate, aerosol, liquid, and water samples, and but are not suitable for determining viability. These
procedures should also be used for detection and viability assessment of rotavirus (Group A) in solid,
particulate, aerosol, liquid, and water samples, although rotavirus is not likely to be viable in aerosol
samples. Note: Procedures described in the USEPA Manual of Methods for Virology (EPA/600/4-
84/013, April 2001) can be adapted for the preparation of samples.
Pathogen(s)
Picornaviruses: Enteroviruses
Picornaviruses: Hepatitis A virus (HAV)
Reoviruses: Rotaviruses (Group A)
Agent Category
Viruses
Viruses
Viruses
Procedures are described for analysis of water samples and may be adapted for assessment of solid,
particulate, aerosol, and liquid samples. The method is used to detect human enteric viruses
(enteroviruses, HAV, rotavirus) in ground water samples. It is a multiplex reverse-transcription
polymerase chain reaction (RT-PCR) procedure optimized for the simultaneous detection of
enteroviruses, HAV, reoviruses, and rotaviruses. Water samples are collected by filtration and viruses are
eluted using a beef extract solution (1.5%, pH 9.5). Viruses are concentrated using celite adsorption (pH
4.0), filtration, and celite-elution with sodium phosphate (0.15 M, pH 9.0), followed by further
concentration and processing to remove inhibitors (ultracentrifugation, solvent extraction, and molecular
weight (MW)-exclusion filtration). Concentrated samples are analyzed by a two-step multiplex RT-PCR
using virus-specific primer sets. Detection of amplicons is by gel electrophoresis with subsequent
confirmation by hybridization (dot-blot) using digoxigenin-labeled internal (nested) probes.
Please note: These procedures have been applied to analysis of ground water. Further research is needed
to develop and standardize the protocols for other sample types. At a minimum, the following quality
control checks should be performed and evaluated before using this protocol: positive control, negative
control, and blank. PCR quality control checks should be performed according to EPA Draft Quality
Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on Environmental
Samples document (www.epa.gov/nerlcwww/qa_qc_pcrl 0_04.pdf) or consult the point of contact
identified in Section 4.
Source: Fout, G. S., Martinson, B. C., Moyer, M. W. N., and Dahling, D. R. 2003. "A Multiplex Reverse
Transcription-PCR Method for Detection of Human Enteric Viruses in Groundwater." Applied and
Environmental Microbiology. 69(6): 3158—3164. (http://aem .asm .org/cgi/reprint/69/6/315 8 .pdf)
7.2.25 Literature Reference for Cryptosporidium spp. (Applied and Environmental
Microbiology. 1999. 65(9): 3936-3941)
These procedures should be used for detection and viability assessment of Cryptosproidium spp. in solid,
)articulate, liquid, and water samples.
Pathogen(s)
Cryptosporidium spp. [Cryptosporidiosis]
Agent Category
Protozoa
Procedures are described for analysis of animal samples and may be adapted for assessment of solid,
particulate, liquid, and water samples. A cell culture infectivity assay capable of detecting infectious
oocysts is used to quantify viable oocysts through sporozoite invasion and clustering of foci. Oocysts
diluted in a standard 5- or 10-fold multiple dilution format are inoculated onto human ileocecal
adenocarcinoma (HCT-8) cell monolayers. After incubation for 48 hours, anti-sporozoite polyclonal
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antibody is used to detect sporozoite invasion, and microscopy is used to confirm replication (life stages
present). Levels of infection and clustering are used to determine the most probable number (MPN) of
infectious oocysts in the stock suspension. For oocysts less than 30 days of age, the correlation between
the initial oocyst inoculum and the MPN calculation is 0.9726. The relationship between the oocyst
inoculum and the MPN diverge as the oocysts age. The 50% infective dose (ID50) in the cell culture
system is approximately 10 oocysts.
Please note: These procedures have been applied to analysis of animal samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: Slifko, T.R., Huffman, D.E., and Rose, J.B. 1999. "A Most-Probable-Number Assay for
Enumeration of Infectious Cryptosporidium parvum Oocysts." Applied and Environmental
Microbiology. 65(9): 3936—3941. http://acm.asm.org/cgi/rcprint/65/9/3936
7.2.26 Literature Reference for Entamoeba histolytica (Journal of Parasitology. 1972.
58(2): 306-310)
These procedures should be used for detection and viability assessment of Entamoeba histolytica in
solid, particulate, liquid, and water samples.
Pathogen(s)
Entamoeba histolytica
Agent Category
Protozoa
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, liquid, and water samples. Entamoeba histolytica cysts are placed in a modified TP-S-1
medium and incubated for 10 hours. Live amoebae excyst through a rupture in the cyst wall, whereas
non-viable amoebae remain encysted. Microscopic examination of an aliquot of the incubated
excystation culture allows calculation of the percent of empty (live) cysts and full (dead) cysts in a
population.
Please note: These procedures have been applied to analysis of clinical (fecal) samples. Further research
is needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: Stringert, R.P. 1972. "New Bioassay System for Evaluating Percent Survival of Entamoeba
histolytica Cysts." The Journal of Parasitology. 53(2): 306—310.
7.2.27 Literature Reference for Entamoeba histolytica (Journal of Clinical Microbiology.
2005. 43(11): 5491-5497)
These procedures should be used for detection of Entamoeba histolytica in solid, particulate, liquid, and
water samples, but are not suitable for determining viability.
Pathogen(s)
Entamoeba histolytica
Agent Category
Protozoa
Procedures are described for analysis of clinical samples and may be adapted for assessment of solid,
particulate, liquid, and water samples. The method is a real-time polymerase chain reaction (PCR) assay
that targets the 18S rRNA gene sequence of E. histolytica. Deoxyribonucleic acid (DNA) is extracted
using cell disruption and a commercial DNA extraction kit, with a second commercial kit used to remove
potential PCR inhibitors. TaqMan® real-time PCR is performed on the purified product. The limit of
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detection is 1 (± 4) cells per mL of sample within 4 hours. The method differentiates between E.
histolytica and E. dispar.
Please note: These procedures have been applied to analysis of clinical (fecal and liver abcess) samples.
Further research is needed to develop and standardize the protocols for other sample types. At a
minimum, the following quality control checks should be performed and evaluated before using this
protocol: positive control, negative control, and blank. PCR quality control checks should be performed
according to EPA Draft Quality Assurance/Quality Control Guidance for Laboratories Performing PCR
Analyses on Environmental Samples document (wjvw^c|M4?OT/ncr^^ or consult
the point of contact identified in Section 4.
Source: Qvarnstrom, Y., James, C., Xayavong, M., Holloway, B.P., Visvesvara, G.S., Sriram, R., and da
Silva, A.J. 2005. "Comparison of Real-time PCR Protocols for Differential Laboratory Diagnosis of
Amebiasis." Journal of Clinical Microbiology. 43(11): 5491-5497.
7.2.28 Literature Reference for Giardia spp. (Transactions of the Royal Society of
Tropical Medicine and Hygiene. 1983. 77(4): 487-488)
These procedures should be used for detection and viability assessment of Giardia spp. in solid,
particulate, liquid, and water samples.
Pathogen(s)
Giardia spp.
Agent Category
Protozoa
Procedures are described for analysis of cell culture samples and may be adapted for assessment of solid,
particulate, liquid, and water samples. TYI-S-33 medium supplemented with bovine bile and additional
cysteine is used to isolate and culture Giardia lamblia. G. lamblia is incubated for intervals of 72 and 96
hours at 36°C in borosilicate glass tubes. The cells form a dense, adherent monolayer on the surface of
the glass.
Please note: These procedures have been applied to analysis of cell culture samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: Keister, D. 1983. "Axenic Culture of Giardia lamblia in TYI-S-33 Medium Supplemented with
Bile." Transactions of the Royal Society of Tropical Medicine and Hygiene. 77(4): 487—488.
7.2.29 Literature Reference for Toxoplasma gondii (Emerging Infectious Diseases. 2006.
12(2): 326-329)
These procedures should be used for detection and viability assessment of Toxoplasma gondii in solid,
particulate, liquid, and water samples.
Pathogen(s)
Toxoplasma gondii [Toxoplasmosis]
Agent Category
Protozoa
Procedures are described for analysis of water samples and may be adapted for assessment of solid,
particulate, liquid, and water samples. Water samples are filtered through fluoropore membrane filters
and concentrated by centrifugation. The filters can be assayed by any of three methods. The first method
involves performing a bioassay in T. go«d//-seronegative chickens. Serum samples are tested by enzyme-
linked immunosorbent assay (ELISA) and/or modified agglutination test until seroconversion, with the
organs from seropositive animals examined microscopically for T. gondii. Mice are injected with brain
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and heart tissue of seropositive chickens, with parasites found in the lungs of mice being confirmatory for
T. gondii. The second method is a similar bioassay with pigs and cats. For the third assay,
deoxyribonucleic acid (DNA) is extracted from the fluoropore membranes for polymerase chain reaction
(PCR) identification of isolates.
Please note: These procedures have been applied to analysis of water samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: de Moura, L., Bahia-Oliveira, L.M.G., Wada, M.Y., Jones, J.L., Tuboi, S.H., Carmo, E.H.,
Ramalho, W.M., Camargo, N.J., Trevisan, R., Graca, R.M.T., da Silva, A.J., Moura, I., Dubey, J.P., and
Garrett, D.O. 2006. "Waterborne Toxoplasmosis, Brazil, from Field to Gene." Emerging Infectious
Diseases. 12(2): 326-329. littp://www.cdc.gov/ncidod/EID/voll2no02/04-1115.htiii
7.2.30 Literature Reference for Toxoplasma gondii (Applied and Environmental
Microbiology. 2004. 70(7): 4035-4039)
These procedures should be used for detection of Toxoplasma gondii in solid, particulate, liquid, and
water samples, but are not suitable for determining viability.
Pathogen(s)
Toxoplasma gondii [Toxoplasmosis]
Agent Category
Protozoa
Procedures are described for analysis of water samples and may be adapted for assessment of solid,
particulate, and liquid samples. The method uses a fluorogenic 5' nuclease (TaqMan®) real-time
polymerase chain reaction (PCR) assay for the detection of T. gondii oocyst deoxyribonucleic acid (DNA)
using gene-specific (Bl gene) primers and probe. Water samples (10 to 100 L) are filtered to concentrate
oocysts. Filters are eluted and recovered oocysts are further purified and concentrated by differential
flotation and centrifugation. Final sample pellets are split and subjected to PCR detection and mouse
bioassay. In experimental seeding assays, a parasite density of 10 oocysts/L is detectable in 100% of the
cases, and a density of 1 oocyst/L is observable in 60% of the cases.
Please note: These procedures have been applied to analysis of water samples. Further research is
needed to develop and standardize the protocols for other sample types. At a minimum, the following
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. PCR quality control checks should be performed according to EPA Draft
Quality Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on
Environmental Samples document (www.epa.gov/nerlcwww/qa_qc_pcr 10_04.pdf) or consult the point of
contact identified in Section 4.
Source: Villena, I., Aubert, D., Gomis, P., Ferte, H., Inglard, J-C., Denise-Bisiaux, H., Dondon, J-M.,
Pisano, E., Ortis, N., and Pinon, J.M. 2004. "Evaluation of a Strategy for Toxoplasma gondii Oocyst
Detection in Water." Applied and Environmental Microbiology. 70(7): 4035—4039.
http://\w\'w.pubmcdccntral .nih.gov/picrcndcr.fcgi ?artid=444816&blobtypc=pdf
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Section 8.0: Selected Biotoxin Methods
A list of methods or procedures to be used in analyzing environmental samples for biotoxin contaminants
is provided in Appendix D. These methods should be used to support remediation activities (site
assessment through clearance) following a homeland security event. Procedures have been compiled for
each biotoxin that may need to be identified and/or quantified following a contamination incident.
Analytical procedures are not currently available for all the analyte-sample type combinations included in
this document. Future research needs include identification of additional methods and development and
validation of the procedures listed. Appendix D is sorted alphabetically by analyte, within each of two
analyte types (i.e., protein and small molecule).
Please note: This section provides guidance for selecting biotoxin methods that have a high likelihood
of assuring analytical consistency when laboratories are faced with a large-scale environmental
restoration crisis. Not all methods have been verified for the analyte/sample type combination listed in
Appendix D. Please refer to the specified method to identify analyte/sample type combinations that
have been verified. Any questions regarding information discussed in this section should be addressed
to the appropriate contact(s) listed in Section 4.
Appendix D provides the following information:
• Analyte(s). The compound or compound(s) of interest.
Analysis type. Tests are either for presumptive identification, confirmatory identification, or
biological activity determination.
• Analytical Technique. An analytical instrument or technique used to determine the quantity and
identification of compounds or components in a sample.
Analytical Method. The recommended method or procedure, and the corresponding publisher.
• Aerosol (filter/cassette or liquid impinger). The recommended method/procedure to measure the
analyte of interest in air sample collection media such as filter cassettes and liquid impingers.
Solid (soil, powder). The recommended method/procedure to measure the analyte of interest in solid
samples such as soil and powders.
• Particulate (swabs, wipes, filters). The recommended method/procedure to measure the analyte of
interest in particulate sample collection media such as swabs, wipes and high efficiency particulate air
(HEPA) filters.
Liquid/drinking water. The recommended method/procedure to measure the analyte of interest in
liquid and drinking water samples.
Some of the biotoxins addressed in this document are commonly found in the environment, and the
methods listed in Appendix D assume that analysis will be used to evaluate contamination levels that are
above background conditions. If possible, an investigation of initial background levels, as well as
controls for background levels, should be performed.
Procedures listed in Appendix D for protein biotoxins are intended to address presumptive, confirmatory,
and biological activity determinations. Because the confirmatory procedures listed for the small molecule
biotoxins involve a determination of intact compound structure (an indication of biological activity
capability), only presumptive and confirmatory methods are listed for these biotoxins. In terms of this
document, presumptive methods, or methods that support a reasonable basis for accurate results, should
be used in situations that require a large number of samples to be processed. Most of the presumptive
methods listed in Appendix D use the immunoassay technique and are designed for large scale sample
processing. The confirmatory method, or the method that corroborates the presumptive results, should be
used on the smaller subset of samples for which presumptive analysis indicates the presence of the
biotoxin. A variety of techniques are listed in Appendix D as confirmatory and generally are more time
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consuming and expensive. The use of these terms in this document is not intended to redefine LRN usage
of these terms. The terms presumptive and confirmatory as used by the LRN are described in Section
8.2.1. If it is necessary to determine the biological activity of a toxin, either an assay (for proteins) or a
technique such as HPLC that determines whether the structure of the biotoxin is intact and likely to be
biologically active (for small molecules) may be used. Biological activity analysis should be applied on
an as-needed basis following analysis with the confirmatory technique.
EPA's NHSRC is working on a sample collection document that is intended as a companion to SAM.
This sample collection document will provide information regarding sampling container/media,
preservation, holding time, sample size, and shipping and is intended to complement the laboratory
analytical methods that are the focus of the SAM document.
8.1 General Guidance
This section provides a general overview of how to identify the appropriate method(s) for a given
biotoxin as well as recommendations for quality control procedures.
For additional information on the properties of the biotoxins listed in Appendix D, TOXNET
Qittgy/toxncyilnijiih.goWindcjyibtil), a cluster of databases on toxicology, hazardous chemicals, and
related areas maintained by the National Library of Medicine, is an excellent resource.
Additional resources include:
A U.S. Army Medical Research Institute of Infectious Diseases' document at
httpi//w¥w.,Maj3™ contains information regarding
sample collection, toxin analysis and identification, as well as decontamination and water treatment.
• Public Health - Select Agents and Toxins, 42 CFR Part 73, found at
The U.S. Centers for Disease Control has information regarding toxins, including 42 CFR Part 1003
regulations for possession, use, and transfer of select agents and toxins, on the following Web site:
Syracuse Research Corporation's Physprop and Chemfate, part of the Environmental Fate Database
supported by EPA. See http://www.syrres.com/esc/databases.htm.
• INCHEM at ht^i//www.mchcm.org/ contains both chemical and toxicity information.
The Registry of Toxic Effects of Chemical Substances (RTECS) database can be accessed via the
NIOSH Web site at http://www.cdc.gov/niosh/rtecs/vz72d288.htmltfJWlDAW for toxicity
information.
EPA's Integrated Risk Information System (IRIS): : //www .epa.gov/i ri s/ contains toxicity
information.
• The Forensic Science and Communications Journal published by the Laboratory Division of the
Federal Bureau of Investigation. See http://www.fbi.gov/hq/lab/fsc/currcnt/backissu.litm.
Additional research on biotoxin contaminants is ongoing within EPA.
8.1.1 Standard Operating Procedures for Identifying Biotoxin Methods
To determine the appropriate method that is to be used on an environmental sample, locate the biotoxin of
concern in Appendix D: Biotoxin Methods under the "Analyte(s)" column. After locating the biotoxin,
continue across the table and identify the appropriate analysis type. After an analysis type has been
chosen, find the analytical technique (e.g., immunoassay) and analytical method applicable to the sample
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type of interest (solid, participate, liquid/drinking water, or aerosol) corresponding to that particular
analyte.
Sections 8.2.1 through 8.2.27 below provide summaries of the analytical methods listed in Appendix D.
Once a method has been identified in Appendix D, Table 8-1 can be used to locate the method summary.
Table 8-1. Biotoxin Methods and Corresponding Text Section Numbers
Biotoxin
Method
Section
Proteins
Abrin
Botulinum neurotoxins1
a-Conotoxin
Ricin1
Shiga and Shiga-like toxins
(Stx, Stx-1 , Stx-2)
Staphylococcal
enterotoxins (SEB)1
Staphylococcal
enterotoxins (SEA, SEC)
119th AOAC Annual Meeting & Exposition, 2005, p. 61 3
Pharmacology & Toxicology. 2001. 88(5): 255-260
Analytical Biochemistry. 2006. 357(2): 200-207
U.S. FDA, Bacteriological Analytical Manual Online, January
2001, Chapter 17, Clostridium botulinum
Lateral Flow Immunoassay Kits
Biochemical Journal. 1997. 328: 245-250
Journal of Medicinal Chemistry. 2004. 47(5): 1234-1241
Analytical Biochemistry. 2006. 357(2): 200-207
Journal of Food Protection. 2005. 68(6): 1294-1301
Lateral Flow Immunoassay Kits
U.S. FDA, Bacteriological Analytical Manual Online, January
2001, Appendix 1, Rapid Methods for Detecting Foodborne
Pathogens
Pharmacology & Toxicology. 2001. 88(5): 255-260
Journal of Clinical Microbiology. 2007. 45(10) 3377-3380
AOAC Official Method 993.06
AOAC Official Method 993.06
8.2.7
8.2.8
8.2.9
8.2.2
8.2.27
8.2.10
8.2.11
8.2.9
8.2.12
8.2.27
8.2.3
8.2.8
8.2.24
8.2.5
8.2.5
Small Molecules
Aflatoxin (Type B1)
a-Amanitin
Anatoxin-a
Brevetoxins (B form)
Cylindrospermopsin
Diacetoxyscirpenol (DAS)
Microcystins (Principal
isoforms: LR, YR, RR, LW)
Picrotoxin
Saxitoxin (STX, NEOSAX,
GTX, dcGTX, dcSTX)
AOAC Official Method 991.31
AOAC Official Method 994.08
Journal of Food Protection. 2005. 68(6): 1294-1 301
Journal of Chromatography. 1991. 563(2): 299-311
Biomedical Chromatography. 1996. 10: 46-47
Environmental Health Perspectives. 2002. 110(2): 179-185
Toxicon. 2004. 43(4): 455-465
FEMS Microbiology Letters. 2002. 216: 159-164
International Journal of Food Microbiology. 1988. 6(1): 9-17
Rapid Communications in Mass Spectrometry. 2006. 20(9):
1422-1428
Journal of AOAC International. 2001. 84(4): 1035-1044
Analyst. 1994. 119(7): 1525-1530
Journal of Pharmaceutical and Biomedical Analysis. 1989. 7(3):
369-375
Journal of AOAC International. 1995. 78: 528-532
8.2.4
8.2.6
8.2.12
8.2.13
8.2.14
8.2.15
8.2.16
8.2.17
8.2.18
8.2.19
8.2.20
8.2.21
8.2.22
8.2.23
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Biotoxin
T-2 Mycotoxin
Tetrodotoxin
Method
Journal of Food Protection. 2005. 68(6): 1294-1301
Rapid Communications in Mass Spectrometry. 2006. 20(9):
1422-1428
Journal of Clinical Laboratory Analysis. 1992. 6: 65-72
Analytical Biochemistry. 2001. 290: 10-17
Section
8.2.12
8.2.19
8.2.25
8.2.26
1For solid, particulate, liquid, and water sample types, LRN methods are used. See Section 8.2.1.
Method summaries are listed in order of method selection hierarchy (see Figure 2-1), starting with EPA
methods followed by methods from other federal agencies, voluntary consensus standard bodies (VCSB),
and journal articles. Methods are listed in numerical order under each publisher. Where available, a
direct link to the full text of the method is provided with the method summary. For additional
information on sample preparation procedures and methods available through consensus standards
organizations, please use the contact information provided in Table 8-2.
Table 8-2. Sources of Biotoxin Methods
Name
U.S. FDA, Bacteriological Analytical
Manual Online, Chapter 17
U.S. FDA, Bacteriological Analytical
Manual Online, Appendix 1
Official Methods of Analysis of AOAC
International*
Pharmacology & Toxicology*
Analytical Biochemistry*
Biochemical Journal
Journal of Medicinal Chemistry*
Journal of Food Protection*
Journal of Chromatography*
Biomedical Chromatography*
Environmental Health Perspectives*
Toxicon*
FEMS Microbiology Letters*
International Journal of Food
Microbiology*
Rapid Communications in Mass
Spectrometry *
Journal of AOAC International*
Publisher
U.S. Food and Drug
Administration
U.S. Food and Drug
Administration
AOAC International
Blackwell Synergy
Science Direct
Biochemical Journal
American Chemical Society
International Association for
Food Protection
Elsevier Science Publishers
John Wiley And Sons Ltd
National Institute of
Environmental Health
Sciences
Elsevier Science Publishers
Blackwell Publishing
Elsevier Science Publishers
John Wiley And Sons Ltd.
AOAC International
Reference
http://www.cfsan.fda.qov/~ebam/bam-
rrjitmi
http://www.cfsan.fda.qov/~ebam/bam-
alhtml
http://www.aoac.org
http://www.blackwell-svnerqy.com/
http://www.sciencedirect.com/
http://www.biochemj.org/
http://www.acs.orq/
http://www.foodprotection.orq/
http://www.wilev.com/
http://www.niehs.nih.qov/
http://www.elsevier.com/
http://www.blackwellpublishinq.com/
http://www.elsevier.com/
http://www.wileY.com/
http://www.aoac.org
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Name
Analyst*
Journal of Pharmaceutical and
Biomedical Analysis*
Journal of Clinical Microbiology
Journal of Clinical Laboratory Analysis*
Lateral Flow Immunoassay Kits
Publisher
Royal Society of Chemistry
Elsevier Science Publishers
American Society for
Microbiology
John Wiley And Sons Ltd.
Environmental Technology
Verification (ETV) Program
Reference
http://www.rsc.orq/
http://www.elsevier.com/
http://www.asm.orq/
http://www.wiley.com/
http://www.epa.qov/etv/
' Subscription and/or purchase required.
8.1.2 General Quality Control (QC) Guidance for Biotoxin Methods
Having data of known and documented quality is critical for public officials to determine the activities
that may be needed during environmental remediation following emergency events. Having such data
requires that laboratories: (1) conduct the necessary QC to ensure that measurement systems are in control
and operating properly, (2) properly document results of the analyses, and (3) properly document
measurement system evaluation of the analysis-specific QC. Ensuring data quality also requires that
laboratory results are properly evaluated and the results of the data quality evaluation are transmitted to
decision makers.
The level or amount of QC needed often depends on the intended purpose of the data that are generated.
Various levels of QC may be required if the data are generated during presence/absence determinations
versus confirmatory analyses. The specific needs for data generation should be identified, and quality
control requirements and data quality objectives should be derived based on those needs, and should be
applied consistently across laboratories when multiple laboratories are used. For example, during rapid
sample screening, minimal QC samples (e.g., blanks, duplicates) and documentation might be required to
ensure data quality. Sample analyses for environmental evaluation during site assessment through site
clearance, such as those identified in this document, might require increased QC (e.g., demonstrations of
method sensitivity, precision, and accuracy).
While method-specific QC requirements may be included in many of the procedures that are cited in this
document, and will be referenced in any standardized analytical protocols developed to address specific
analytes and sample types of concern, the following describes a minimum set of QC samples and
procedures that should be conducted for all analyses. Individual methods, sampling and analysis
protocols, or contractual statements of work also should be consulted to determine any additional QC that
may be needed. QC tests should be run as frequently as necessary to ensure the reliability of analytical
results. In general, sufficient QC includes an initial demonstration of measurement system capability as
well as ongoing assessments to ensure the continued reliability of the analytical results.
Examples of sufficient quality control for the presumptive tests listed in Appendix D include:
Method blanks;
• Positive test samples / negative test samples;
• Calibration check samples;
• Use of test kits and reagents prior to expiration; and
Accurate temperature controls.
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Examples of sufficient quality control for the confirmatory tests listed in Appendix D include:
• Demonstration that the measurement system is operating properly
*• Initial calibration
*• Method blanks
• Demonstration of measurement system suitability for intended use
*• Precision and recovery (verify measurement system has adequate accuracy)
*• Analyte/sample type/level of concern-specific QC samples (verify that measurement system has
adequate sensitivity at levels of concern)
Demonstration of continued measurement system reliability
*• Matrix spike/matrix spike duplicates (recovery and precision)
*• QC samples (system accuracy and sensitivity at levels of concern)
*• Continuing calibration verification
*• Method blanks
Please note: The appropriate point of contact identified in Section 4 should be consulted regarding
appropriate QA/QC procedures prior to sample analysis. These contacts will consult with the EPA
OSWER coordinator responsible for laboratory activities during the specific event to ensure QA/QC
procedures are performed consistently across laboratories. OSWER is planning to develop QA/QC
guidance for laboratory support. EPA program offices will be responsible for ensuring that the QA/QC
practices are implemented.
8.1.3 Safety and Waste Management
It is imperative that safety precautions be used during collection, processing, and analysis of
environmental samples. Laboratories should have a documented health and safety plan for handling
samples that may contain target chemical, biological, or radiological contaminants, and laboratory staff
should be trained in and implement the safety procedures included in the plan. In addition, many of the
methods summarized or cited in Section 8.2 contain some specific requirements, guidance, or information
regarding safety precautions that should be followed when handling or processing environmental samples
and reagents. These methods also provide information regarding waste management. Other resources
that can be consulted for additional information include the following:
• Occupational Health and Safety Administration's (OSHA) standard for Occupational Exposure to
Hazardous Chemicals in Laboratories (29 CFR 1910.1450);
• OSHA regulations for hazardous waste operations and emergency response (29 CFR 1910.120);
• Environmental Protection Agency's standards regulating hazardous waste (40 CFR Parts 260 - 270);
• U.S. Department of Transportation (DOT) regulations for transporting hazardous materials (49 CFR
Part 172);
• U.S. Department of Health and Human Services, Centers for Disease Control and Prevention's
requirements for possession, use, and transfer of select agents and toxins (42 CFR Part 1003);
• American Society for Microbiology, Biological Safety: Principles and Practices, 4th Ed.
(http://cstorc.asm.org/); and
• American Biological Safety Association, Risk Group Classifications for Infectious Agents
(http://www.absa.org/resriskgroup.html).
• Select Agent Rules and Regulations (42 CFR Part 73 and 9 CFR Part 121) found at
02,tol
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8.2 Method Summaries
Summaries for the analytical methods listed in Appendix D are provided in Sections 8.2.1 through 8.2.27.
These sections contain summary information only, extracted from the selected methods. The full version
of the method should be consulted prior to sample analysis.
Each summary contains a table identifying the biotoxin(s) and sample type to which the method applies, a
brief description of the method, performance data (if available), and a link to or source for obtaining a full
version of the method.
8.2.1 Laboratory Response Network (LRN)
The agents and sample types identified below and listed in Appendix D are included in the U.S.
Department of Health and Human Services/U.S. Department of Agriculture (HHS/USDA) select agent list
and should be analyzed in accordance with the appropriate LRN protocols.
The LRN was created in accordance with Presidential Directive 39, which established terrorism
preparedness responsibilities for federal agencies. The LRN is primarily a national network of local,
state, federal, military, food, agricultural, veterinary, and environmental laboratories; however, additional
LRN laboratories are located in strategic international locations. The Centers for Disease Control and
Prevention (CDC) provides technical and scientific support to member laboratories as well as secure
access to standardized procedures and reagents for rapid (within 4 to 6 hours) presumptive detection of
biothreat agents and emerging infectious disease agents. These rapid presumptive assays are part of
agent-specific algorithms of assays which lead to a confirmed result. The algorithm for a confirmed
result is often a combination of one or more presumptive positive results from a rapid assay in
combination with a positive result from one of the "gold standard" methods, such as culture. The
standardized procedures, reagents, and agent-specific algorithms are considered to be sensitive and are
only available to LRN member laboratories. Thus, these procedures are not available to the general
public and are not discussed in this document.
It is important to note that, in some cases, the procedures may not be fully developed or validated for each
environmental sample type/analyte combination listed in Appendix D, nor are all LRN member
laboratories necessarily capable of analyzing all of the sample type/analyte combinations.
Analyte(s)
Botulinum neurotoxins (Serotypes A, B, E, F)
Ricin toxin
Staphylococcal enterotoxin B (SEB)
Sample Type
Solid, Participate, Liquid/Water
Solid, Participate, Liquid/Water
Solid, Particulate, Liquid/Water
CASRN
NA
9009-86-3
39424-53-8
Please note: Not all methods have been verified for the biotoxin/sample type combination listed in
Appendix D. Please refer to the agent-specific method to identify the biotoxin/sample type combinations
that have been validated. Any questions regarding information discussed in this section should be
referred to the appropriate contact(s) listed in Section 4.
For additional information on the LRN, including selection of a laboratory capable of receiving and
processing the specified sample type/analyte, please use the contact information provided below or visit
http://www.bt.cdc.gov/lrii/.
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Centers for Disease Control and Prevention
Laboratory Response Branch
Division of Bioterrorism Preparedness and Response (DBPR)
National Center for Prevention, Detection, and Control of Infectious Diseases (NCPDCID)
Coordinating Center for Infectious Diseases (CCID)
Centers for Disease Control and Prevention (CDC)
1600 Clifton Road NE, Mailstop C-18
Atlanta, GA 30333
Telephone: (404) 639-2790
E-mail: lm@cdc.gov
Local public health laboratories, private, and commercial laboratories with questions about the LRN
should contact their state public health laboratory director or the Association of Public Health
Laboratories (contact information provided below).
Association of Public Health Laboratories
8515 Georgia Avenue, Suite 700
Silver Spring, MD, 20910
Telephone: (240) 485-2745
Fax: (240) 485-2700
Web site: ww,w,gphLorg
E-mail: info@aphl.org
8.2.2 U.S. FDA, Bacteriological Analytical Manual Online, Chapter 17, 2001: Botulinum
Neurotoxins
These methods should be used for the confirmatory and biological activity analysis of Types A, B, E,
and F botulinum neurotoxins in aerosol samples.
Analyte(s)
Botulinum neurotoxins
(Serotypes A, B, E, F)
Agent Category
Protein
CASRN
NA
An amplified-enzyme-linked immunosorbent assay (amp-ELISA) and a digoxigenin-labeled enzyme-
linked immunosorbent assay (DIG-ELISA) are described for the detection of Types A, B, E, and F
botulinum neurotoxins in food products. The amp-ELISA method uses goat anti-A or E, rabbit anti-B, or
horse anti-F serum to capture the toxins in a 96-well plate, and a corresponding biotinylated goat antitoxin
to detect the toxin. Visualization is with streptavidin-alkaline phosphatase. The DIG-ELISA method is a
modification of the amp-ELISA method, with digoxigenin-labeled antitoxin IgG's substituted for the
streptavidin-alkaline phosphatase. Toxin can be detected at approximately 10 minimum lethal doses
(MLD)/mL (0.12 to 0.25 ng/mL). Very toxic samples (greater than 10,000 MLD/mL) may give a positive
absorbance for more than one toxin type. Further dilution of the sample will remove cross-reactivity.
The mouse bioassay detects biologically active toxin using a three part approach: toxin screening; toxin
titer; and finally, toxin neutralization using monovalent antitoxins. Samples are prepared by
centrifugation for suspended solids under refrigeration, or solids are extracted with an equal volume of pH
6.2 gel-phosphate buffer and then centrifuged. Toxins of nonproteolytic types may need trypsin
activation to be detected. Serial dilutions of untreated and trypsin-treated sample fluids are injected in
separate pairs of mice intraperitoneally (i.p.). Mice are also injected with heated untreated, undiluted
sample. Death of mice, along with symptoms of botulism, confirms presence of botulinum toxin. After
calculation of an MLD, dilute monovalent antitoxins types A, B, E, and F are injected into mice 30
minutes to 1 hour before challenging them with the i.p. injection of each dilution that gave the highest
MLD from the toxic preparation.
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Please note: This method has been applied to analysis of food products. Further research is needed to
develop and standardize the protocols for other sample types.
Source: U.S. Food & Drug Administration, Center for Food Safety & Applied Nutrition. 2001.
Bacteriological Analytical Manual Online. Chapter 17. (http://www.cfsan.fda.gov/~cbam/bam-17.html)
8.2.3 U.S. FDA, Bacteriological Analytical Manual Online, Appendix 1, 2001: Rapid
Methods for Detecting Foodborne Pathogens
This method should be used for the presumptive analysis of Shiga (Stx) and Shiga-like (Stx-1 and Stx-2)
biotoxins in aerosol, solid, particulate, liquid, and water samples.
Analyte(s)
Shiga and Shiga-like toxins
(Stx, Stx-1 , Stx-2)
Agent Category
Protein
CASRN
75757-64-1
Shiga toxin (Stx) is produced by Shigella dysenteriae and Shiga-like toxins (Stx-1 and Stx-2) are
produced by various Shiga-toxigenic E. coll (STEC). An enzyme-linked immunosorbent assay (ELISA)
is described for the detection of these toxins. Diluted samples are added to microwells coated with an
anti-Shiga toxin capture antibody. After incubation at room temperature, a wash is performed to remove
unbound material. A second anti-Shiga toxin antibody is added for detection and incubation continued at
room temperature. A wash is performed to remove unbound antibody. Enzyme conjugated anti-IgG
visualization antibody is added and the plate incubated then rinsed. Substrate is added, and after
incubation to develop the color, stop solution is added. The results are interpreted
spectrophotometrically.
Please note: This method has not been applied to analysis of environmental samples. Further research is
needed to develop and standardize the protocols for these sample types.
Source: U.S. Food & Drug Administration, Center for Food Safety & Applied Nutrition. 2001.
Bacteriological Analytical Manual Online. Appendix 1. (http://www.cfsan.fda.gov/~ebam/bam-al.html)
8.2.4 AOAC Official Method 991.31: Aflatoxins in Corn, Raw Peanuts, and Peanut Butter
This method should be used for presumptive analysis of aflatoxin Type Bl in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Aflatoxin (Type B1)
Agent Category
Small Molecule
CASRN
1402-68-2
This method is for the detection of aflatoxins in agricultural products. The sample is extracted with
methanol-water (7 + 3), filtered, diluted with water, and applied to an affinity column containing
monoclonal antibody specific for aflatoxins Bl, B2, Gl, and G2. Antibody-bound aflatoxins are removed
from the column with methanol. Total aflatoxins are quantified by fluorescence measurement after
reaction with bromine solution. Individual aflatoxins are quantitated by LC with fluorescence detection
and postcolumn iodine derivatization. Method performance was characterized using various commodities
(e.g., corn) at aflatoxin levels over a range of 10 to 30 ng/g. This method was originally designed for the
analysis of aflatoxins (Bi, B2, GI, and G2) in samples where cleanup was necessary to remove food
components, such as fats and proteins; the cleanup procedure may not be necessary for analysis of water
samples.
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Please note: This method has been applied to analysis of agricultural products. Further research is
needed to develop and standardize the protocols for other sample types.
Source: AOAC International. 1994. Official Methods of Analysis ofAOAC International. 16th Edition, 4th
Revision: Vol II.
8.2.5 AOAC Official Method 993.06: Staphylococcal Enterotoxins in Selected Foods
This method should be used for presumptive analysis of Staphylococcal enterotoxins Type B in aerosol
samples, and Types A and C in aerosol, solid, particulate, liquid, and water samples.
Analyte(s)
Staphylococcal enterotoxins
(SEE, SEA, SEC)
Agent Category
Protein
CASRN
39424-53-8, 37337-57-8,
39424-54-9
This method is an enzyme immunoassay (EIA) using a mixture of high-affinity capture for identification
of toxin(s) in food samples. Samples are prepared by dilution in Tris buffer, centrifugation, and filtration
of the supernatant through a syringe, with adjustment to a final pH of 7.0 to 8.0. Samples are incubated in
96-well plates with the mixture of antibodies conjugated to horseradish peroxidase, and visualized with an
acetic peroxidase substrate. Assay results are determined visually or using a microtiter plate reader. Test
is considered positive for Staphylococcal enterotoxins if absorbance is >0.200 and is considered negative
if absorbance is <0.200. Specific toxin serotypes are not differentiated. This method detects from 1.3 to
3.3 ng/mL Staphylococcal enterotoxin in extracts prepared from food containing 4 to 10 ng/mL
staphyloccal enterotoxin.
Please note: This method has been applied to analysis of food products. Further research is needed to
develop and standardize the protocols for other sample types.
Source: AOAC International. 1994. Official Methods of Analysis of AOAC International. 16th Edition, 4th
Revision: Vol I.
8.2.6 AOAC Official Method 994.08: Aflatoxin in Corn, Almonds, Brazil Nuts, Peanuts,
and Pistachio Nuts
This method should be used for confirmatory analysis of aflatoxin Type Bl in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Aflatoxin (Type B1)
Agent Category
Small Molecule
CASRN
1402-68-2
This method is for the identification of aflatoxins in agricultural products. Samples are extracted using an
acetonitrile-water (9 + 1) solution. Sample extracts are then run through a multifunctional cleanup
column. The purified extract and standards are derivatized with trifluoracetic acid, and then analyzed
using a high performance liquid chromatography (HPLC) system with a fluorescence detector. Specific
aflatoxins can be identified by their retention time and quantified using standard curves. Method
performance was characterized using various commodities (e.g., corn) at aflatoxin levels over a range of 5
to 30 ng/g. This method was originally designed for the analysis of aflatoxins (Bi, B2, GI, and G2) in
commodities where cleanup was necessary to remove other food components, such as fats and proteins;
the cleanup procedure may not be necessary for water analyses. Coupling the procedures, or a
modification of the procedures, included in this method with an immunoassay and/or viability test (where
available) will provide more information regarding specificity and toxicity.
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Please note: This method has been applied to analysis of agricultural products. Further research is
needed to develop and standardize the protocols for other sample types.
Source: AOAC International. 1998. Official Methods of Analysis ofAOAC International. 16th Edition, 4th
Revision; Vol II.
8.2.7 Literature Reference for Abrin (119th AOAC Annual Meeting & Exposition, 2005, p.
613)
These procedures should be used for presumptive analysis of abrin in aerosol, solid, particulate, liquid,
and water samples.
Analyte(s)
Abrin
Agent Category
Protein
CASRN
1393-62-0
Procedures are described for using mouse monoclonal antibodies and rabbit derived polyclonal antibodies
prepared against a mixture of abrin isozymes for three separate enzyme-linked immunosorbent assay
(ELISA) and electrochemiluminescence (ECL)- based assays in food products. The three assays vary by
use of antibody combination: (1) polyclonal (capture)/polyclonal (detection) ELISA, (2)
polyclonal/monoclonal ELISA, and (3) polyclonal/monoclonal ECL assay. The limits of detection (LOD)
with purified Abrin C and various abrin extracts in buffer are between 0.1 and 0.5 ng/mL for all three
assays. The LOD for abrin dissolved into food products ranges from 0.1 to 0.5 ng/mL, using the ECL
assay. The LOD for abrin dissolved into food products for the ELISA assays range between 1 and 4
ng/mL, depending on the assay configuration. In all cases, the LODs are considerably less than the
concentration at which abrin may pose a health concern.
Please note: These procedures have been applied to analysis of food products. Further research is needed
to develop and standardize the protocols for other sample types.
Source: Garber, E.A., Aldrich, J.L., Wang, J., Brewer, V.A., O'Brien, T.W., and Sigal, G. 2005.
"Detection of Abrin Foods Using ELISA and Electrochemiluminescence (ECL) Technologies." 779*
AOAC Annual Meeting & Exposition, p. 613.
8.2.8 Literature Reference for Abrin and Shiga and Shiga-like Toxins (Pharmacological
Toxicology. 2001. 88(5): 255-260)
These procedures should be used for confirmatory analysis of abrin and Shiga and Shiga-like toxins in
aerosol, solid, particulate, liquid, and water samples.
Analyte(s)
Abrin
Shiga and Shiga-like Toxins
(Stx, Stx-1 , Stx-2)
Agent Category
Protein
Protein
CASRN
1393-62-0
75757-64-1
Procedures are described for measuring the biological activity of ribosome-inactivating proteins using a
microtiter plate format for detection of abrin in phosphate buffered saline (PBS). Nuclease-treated rabbit
reticulocyte lysate containing luciferase messenger ribonucleic acid (mRNA) is used to measure toxin
activity via inhibition of protein synthesis. The relative biological activity is determined by comparing
luminescence levels in treated samples versus those of untreated controls. The amount of luciferase
translated, as measured by luminescence, is inversely proportional to the toxin concentration. Linear dose
response curves are generated for abrin, with a 50% inhibition of translation at 0.5 nM. Coupling this
procedure, or a modification of this procedure, with an immunoassay will provide more information
regarding the specificity and toxicity of the target biotoxin.
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Please note: These procedures have been applied to analysis of abrin in PBS. Further research is needed
to develop and standardize the protocols for other sample types.
Source: Hale, M.L. 2001. "Microtiter-based Assay for Evaluating the Biological Activity of Ribosome-
inactivation Proteins." Pharmacological Toxicology. 88(5): 255—260.
8.2.9 Literature Reference for Abrin and Ricin (Analytical Biochemistry. 2006. 357(2):
200-207)
These procedures should be used for biological activity analysis of abrin and ricin in aerosol, solid,
particulate, liquid, and water samples.
Analyte(s)
Abrin
Ricin
Agent Category
Protein
Protein
CASRN
1393-62-0
9009-86-3
This assay is an N-glycosidase enzyme activity assay for the detection of purified abrin and ricin in a
reaction buffer consisting of nuclease free water with Triton X-100. Synthetic biotinylated RNA
substrates are cleaved by the combined actions of the toxin and a chemical agent, N,N'-
dimethylethyldiamine. Annealing of the product with a ruthenylated oligodeoxynucleotide results in the
capture of ruthenium chelate onto magnetic beads. Inclusion of the monoclonal antibody (Mab) 9C3 is
used to enhance the N-glycosidase activity. Commercially available electrochemiluminescence (ECL)-
based reagents are used to detect the product. Polyclonal/monoclonal antibodies are commercially
available as an ELISA test kit. The activity assay exhibits similar limits of detection just below signal
with 0.1 ng/ml of ricin; the ECL response was linear as the ricin concentration increased by two orders of
magnitude. Abrin II similarly cleaves the biotinylated RNA substrate, with the N-glycosidase activity of
the toxin enhanced to a greater degree by Mab 9C3, enabling differentiation from ricin.
Please note: These procedures have been applied to analysis of purified toxins in reaction buffer. Further
research is needed to develop and standardize the protocols for other sample types.
Source: Keener, W.K., Rivera, V.R., Young, C.C., and Poli, M.A. 2006. "An Activity-dependent assay
for Ricin and Related RNA N-glycosidases Based on Electrochemiluminescence." Analytical
Biochemistry. 357(2): 200-207.
8.2.10 Literature Reference for a-Conotoxin (Biochemical Journal. 1997. 328: 245-250)
These procedures should be used for presumptive and biological activity analysis of a-conotoxin in
aerosol, solid, particulate, liquid, and water samples.
Analyte(s)
a-Conotoxin
Agent Category
Protein
CASRN
156467-85-5
A biologically active fluorescein derivative of Conus geographus a-conotoxin (FGI) is used in solution-
phase-binding assays with purified Torpedo californica nicotinic acetylcholine receptor (nAchR) and
monoclonal antibodies (Mabs) to detect the toxin in laboratory samples. FGI binding to T. californica
nAchR or Mabs is determined by spin-column at room temperature. For competitive ligand-displacement
assays, FGI is premixed with various dilutions of unlabelled ligands and then incubated with nAchR or
mAb. Fluorescence is measured in ratio mode using cuvettes with excitation and emission
monochromators set at gamma = 490 nm and gamma = 525 nm respectively. The binding of FGI to
nAchR or antibody had apparent dissociation constants of 10 to 100 nM.
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Please note: These procedures have been applied to analysis of purified toxin in phosphate buffer.
Further research is needed to develop and standardize the protocols for other sample types.
Source: Ashcom, J.D., and Stiles, E.G. 1997. "Characterization of a-Conotoxin Interactions with the
Nicotinic Acetylcholine Receptor and Monoclonal Antibodies." BiochemicalJournal. 328: 245—250.
http://www.bioc.hcmi .org/bi/328/0245/3280245 .pdf
8.2.11 Literature Reference for a-Conotoxin (Journal of Medicinal Chemistry. 2004. 47(5):
1234-1241)
These procedures should be used for confirmatory analysis of a-conotoxin in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
a-Conotoxin
Agent Category
Protein
CASRN
156467-85-5
Procedures are discussed for the detection of peptides within the a-conotoxin molecular mass range using
high performance liquid chromatography-mass spectrometry (HPLC-MS). A crude extract of the sample
is made using 30% acetonitrile/water acidified with 0.1% trifluoroacetic acid (TFA), with the insoluble
portion of the sample removed by centrifugation. A portion of the sample extract is fractionated by size-
exclusion chromatography in order to prepare a sample containing small peptides in the range of 1000 to
2500 Da. Chromatography conditions are elution with 30% acetonitrile / 0.048% TFA at a flow rate of
0.5 mL/minute, with detection at 214 nm. Three sulfated alpha-conotoxins (AnIA, AnIB, and AnIC) can
be identified by LC-MS. Peptides can be quantified by reversed-phase high performance liquid
chromatography (RP-HPLC) using an external reference standard for each peptide.
Please note: These procedures have been applied to analysis ofConus anemone venom duct material.
Further research is needed to develop and standardize the protocols for other sample types.
Source: Loughnan, M.L., Nicke, A., Jones, A., Adams, D.J., Alewood, P.P., and Lewis, R.J. 2004.
"Chemical and Functional Identification and Characterization of Novel Sulfated Alpha-conotoxins from
the Cone Snail Conus anemone." Journal of Medicinal Chemistry. 47(2): 1234—1241.
8.2.12 Literature Reference for a-Amanitin, Ricin, T-2 Mycotoxin (Journal of Food
Protection. 2005. 68(6): 1294-1301)
These procedures should be used for presumptive analysis of a-amanitin and T-2 toxin in aerosol, solid,
particulate, liquid, and water samples and for confirmatory analysis of ricin in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
a-Amanitin
Ricin
T-2 Mycotoxin
Agent Category
Small Molecule
Protein
Protein
CASRN
23109-05-9
9009-86-3
21259-20-1
Commercially available enzyme-linked immunosorbent assays (ELISAs) are described and assessed for
detection of ricin, amanitin, and T-2 toxin at levels below those described as a health concern in food
samples. Solid food samples are prepared by washing the sample with sodium phosphate buffer followed
by dilution with phosphate-buffered saline. Liquid beverage samples are prepared by dilution in sodium
phosphate buffer. Amanitin samples are similarly prepared using water instead of buffer, and T-2 toxin
samples are similarly prepared using 35% methanol in water instead of buffer. The prepared samples are
used with commercially obtained ELISA kits according to the manufacturer's directions, except for the
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incorporation of an eight-point calibration curve and reading the plates at both 405 and 650 nm after 26
minutes of incubation at 37°C. This assay detects ricin in food products at 0.01 ug/mL with acceptable
background levels. Amanitin can be detected in food products at 1 ug/g with the ELISA. Background
responses occurred, but at less than the equivalent of 0.5 ppm for amanitin. The ELISA kit will
successfully detect T-2 toxin at targeted levels of 0.2 ug/g. The ELISA kit successfully detects T-2 toxin
at targeted levels of 0.2 ug/g, the immunoassay for T-2 toxin, however; shows significant background
responses and varies up to 0.1 ppm.
Please note: These procedures have been applied to analysis of food products. Further research is needed
to develop and standardize the protocols for other sample types.
Source: Garber, E.A., Eppley, R.M., Stack, M.E., McLaughlin, M.A., and Park, D.L. 2005. "Feasibility
of Immunodiagnostic Devices for the Detection of Ricin, Amanitin, and T-2 Toxin in Food." Journal of
Food Protection. 68(6): 1294-1301.
8.2.13 Literature Reference for a-Amanitin (Journal of Chromatography. 1991. 563(2):
299-311)
These procedures should be used for confirmatory analysis of a-amanitin in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
a-Amanitin
Agent Category
Small Molecule
CASRN
23109-05-9
Procedures are described for the selective determination in human plasma of a-amanitin using high-
performance liquid chromatography (HPLC) with amperometric detection. After extraction of plasma
with disposable Qg silica cartridges, the extracts are separated by isocratic reversed-phase
chromatography using a macroporous polystyrene-divinylbenzene column and a mobile phase of 0.05 M
phosphate buffer-acetonitrile (91:9) at pH 9.5. Amperometric detection is performed by applying an
oxidation potential as low as +350 mV (vs. Ag/AgCl) to a glassy carbon electrode, in a thin-layer flow-
cell. The linear range for alpha-amanitin is 3 to 200 ng/mL, and the relative limit of detection in plasma
is 2 ng/mL at a signal-to-noise ratio of 2. The intra-assay precision has been evaluated at levels of 10 and
200 ng/mL.
Please note: These procedures have been applied to analysis of plasma. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Tagliaro, F., Schiavon, G., Bontempelli, G., Carli, G., and Marigo, M. 1991. "Improved High-
performance Liquid Chromatographic Determination with Amperometric Detection of Alpha-amanitin in
Human Plasma Based on its Voltammetric Study." Journal of Chromatography. 563(2): 299—311.
8.2.14 Literature Reference for Anatoxin-a (Biomedical Chromatography. 1996.10: 46-47)
These procedures should be used for confirmatory analysis of anatoxin-a in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Anatoxin-a
Agent Category
Small Molecule
CASRN
64285-06-9
Procedures are described for high-performance liquid chromatography (HPLC) analysis with fluorimetric
detection of anatoxin-a in water samples after derivatization with 7-fluoro-4-nitro-2,l,3-benzoxadiazole
(NBD-F). Samples are extracted at pH 7 with solid phase extraction (SPE) using a weak cation
exchanger. The toxin is eluted with methanol containing 0.2% trifluoroacetic acid. Samples are
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evaporated, reconstituted with acetonitrile, and re-evaporated prior to derivatization. This procedure
detects anatoxin-a at concentrations of 0.1 ug/L with a good linear calibration.
Please note: These procedures have been applied to analysis of water. Further research is needed to
develop and standardize the protocols for other sample types.
Source: James, K.J., and Sherlock, I.R. 1996. "Determination of the Cyanobacterial Neurotoxin,
Anatoxin-a, by Derivatisation Using 7-Fluoro-4-Nitro-2,l,3-Benzoxadiazole (NBD-F) and HPLC
Analysis with Fluorimetric Detection." Biomedical Chromatography. 10: 46—47.
8.2.15 Literature Reference for Brevetoxins (Environmental Health Perspectives. 2002.
110(2): 179-185)
These procedures should be used for presumptive analysis of brevetoxins in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Brevetoxins (B form)
Agent Category
Small Molecule
CASRN
98225-48-0
Procedures are described for a competitive enzyme-linked immunosorbent assay (ELISA) used to detect
brevetoxins in shellfish. The assay uses goat anti-brevetoxin antibodies in combination with a three-step
signal amplification process: (1) secondary biotinylated antibodies; (2) streptavidine-horseradish
peroxidase conjugate; and (3) chromogenic enzyme substrate. Sample preparation for liquids is dilution
in phosphate buffered saline (PBS). Sample preparation for solids (oysters) is homogenization in PBS, or
extraction in acetone. The working range for the assay is 0.2 to 2 ng/mL for diluted and undiluted liquid
samples, and 0.2 to 2 ng/mL for solid samples, corresponding to 0.8 to 8 ug brevetoxins/100 g shellfish.
The method has been compared to the mouse bioassay and is equivalent in sensitivity.
Please note: These procedures have been applied to analysis of shellfish. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Naar, J., Bourdelais, A., Tomas, C., Kubanek, J., Whitney, P.L., Flewelling, L., Steidinger, K.,
Lancaster, J., and Badan, D.G. 2002. "A Competitive ELISA to Detect Brevetoxins from Karenia brevis
(Formerly Gymnodinium breve) in Seawater, Shellfish, and Mammalian Body Fluid." Environmental
Health Perspectives. 110(2): 179-185.
http://\vw\v.pubniedcentral .nih.gov/picrender.fcgi ?artid=1240733&blobt>'pe=pdf
8.2.16 Literature Reference for Brevetoxins (Toxicon. 2004. 43(4): 455-465)
These procedures should be used for confirmatory analysis of brevetoxins in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Brevetoxins (B form)
Agent Category
Small Molecule
CASRN
98225-48-0
Shellfish sample homogenates are extracted with acetone, and centrifuged. The supernatants are
combined, evaporated, and re-solubilized in 80% methanol. Following a wash with 95% n-hexane, the
methanolic layer is evaporated, and the residue re-solubilized in 25% methanol and applied to a Ci8 SPE
column. Analytes are eluted with 100% methanol, evaporated, and re-solubized in methanol for analysis.
Analysis of prepared samples is performed using liquid chromatography tandem mass spectrometers (LC-
MS-MS) with a mobile phase of water and acetonitrile with acetic acid. Analytes are detected by mass
spectrometer with electrospray ionization (ESI) interface. Brevetoxins are extensively metabolized, with
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many sub-forms. This method describes multiple LC-MS-ESI profiles for metabolites of brevetoxins
from oysters.
Please note: These procedures have been applied to analysis of shellfish. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Wang, Z., Plakas, S.M., El Said, K.R., Jester, E.L., Granade, H.R., and Dickey, R.W. 2004.
"LC/MS Analysis of Brevetoxin Metabolites in the Eastern Oyster (Crassostrea virginica)" Toxicon.
43(4): 455-465.
8.2.17 Literature Reference for Cylindrospermopsin (FEMS Microbiology Letters. 2002.
216(2): 159-164)
These procedures should be used for confirmatory analysis of Cylindrospermopsin in aerosol, solid,
particulate, liquid, and water samples.
Analyte(s)
Cylindrospermopsin
Agent Category
Small Molecule
CASRN
143545-90-8
Cylindrospermopsin is detected using high performance liquid chromatography with photodiode array
detector (HPLC-PDA) in environmental waters. The suggested solvent range for Cylindrospermopsin is
below 50% methanol and 30% acetonitrile. Complex samples (culture medium) are purified on a Ci8
column with a linear gradient of 1 to 12% (v/v) methanol/water over 24 minutes at 40°C, with monitoring
at 262 nm. The use of Qs columns for environmental waters is suggested for removal of the large
number of organic compounds that may be present. This method detects and recovers
Cylindrospermopsin from spiked environmental water samples at 1 ug/L.
Please note: These procedures have been applied to analysis of water. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Metcalf, J.S., Beattie, K.A., Saker, M.L., and Codd, G.A. 2002. "Effects of Organic Solvents on
the High Performance Liquid Chromatographic Analysis of the Cyanobacterial Toxin
Cylindrospermopsin and Its Recovery from Environmental Eutrophic Waters by Solid Phase Extraction."
FEMS Microbiology Letters. 216(2): 159-164.
8.2.18 Literature Reference for Diacetoxyscirpenol (DAS) (International Journal of Food
Microbiology. 1988. 6(1): 9-17)
These procedures should be used for presumptive analysis of diacetoxyscirpenol (DAS) in aerosol, solid,
particulate, liquid, and water samples.
Analyte(s)
Diacetoxyscirpenol (DAS)
Agent Category
Small Molecule
CASRN
2270-40-8
An enzyme-linked immunosorbent assay (ELISA) is used for the detection of diacetoxyscirpenol (DAS)
in food samples. Antibodies against DAS are obtained after immunization of rabbits with DAS-
hemiglutarate-human serum albumin (DAS-HG-HSA), and a DAS-hemisuccinate-horseradish
peroxidase-conjugate (DAS-HS-HRP) is prepared by an ester method for use as enzyme-labeled toxin in
the competitive assay. The detection limit for DAS using this assay is approximately 10 pg/mL. The
relative cross reactivities of the assay are 597.5, 5.2, 100.0, 2.5, and 1.5% for 3 alpha-acetyl-DAS, DAS,
T-2 toxin, neosolaniol, and 15-acetoxyscirpenol, respectively.
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Please note: These procedures have been applied to analysis of food. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Klaffer, U., Martlbauer, E., and Terplan, G. 1988. "Development of a Sensitive Enzyme-linked
Immunosorbent Assay for the Detection of Diacetoxyscirpenol." International Journal of Food
Microbiology. 6(1): 9-17.
8.2.19 Literature Reference for Diacetoxyscirpenol (DAS) and T-2 Mycotoxin (Rapid
Communications in Mass Spectrometry. 2006. 20(9): 1422-1428)
These procedures should be used for confirmatory analysis of diacetoxyscirpenol (DAS) and T-2
mycotoxin in aerosol, solid, particulate, liquid, and water samples.
Analyte(s)
Diacetoxyscirpenol (DAS)
T-2 Mycotoxin
Agent Category
Small Molecule
Small Molecule
CASRN
2270-40-8
21259-20-1
A liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (LC/APCI-MS)
procedure based on time-of-flight mass spectrometry (TOFMS), with a real-time reference mass
correction, is used for simultaneous determination ofFusarium mycotoxins (to include
diacetoxyscirpenol (DAS) and T-2 mycotoxin) in foodstuffs. Mycotoxin samples are extracted with
acetonitrile/water (85:15) and centrifuged, and the supernatant is applied to a column for cleanup.
Prepared samples are separated by liquid chromatography with an aqueous mobile phase of ammonium
acetate and methanol detection is provided in exact mass chromatograms with a mass window of 0.03 Th.
The limits of detection range from 0.1 to 6.1 ng/g in analyzed foodstuffs.
Please note: These procedures have been applied to analysis of food. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Tanaka, H., Takino, M., Sugita-Konishi, Y., and Tanaka, T. 2006. "Development of Liquid
Chromatography/Time-of-flight Mass Spectrometric Method for the Simultaneous Determination of
Trichothecenes, Zearalenone, and Aflatoxins in Foodstuffs." Rapid Communications in Mass
Spectrometry. 20(9): 1422-1428.
8.2.20 Literature Reference for Microcystins (Journal of AOAC International. 2001. 84(4):
1035-1044)
These procedures should be used for presumptive analysis of microcystins in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Microcystins
(Principal Isoforms: LR, YR, RR, LW)
Agent Category
Small Molecule
CASRN
77238-39-2
Enzyme-linked immunosorbent assay (ELISA) and protein phosphatase inhibition assay are used to detect
microcystins in algae products. Solid samples are prepared by homogenization in methanol (75% in
water), with centrifugation to remove solids. Immunoassays are performed on the prepared samples using
a commercially available ELISA test kit as described by the manufacturer. Samples are quantitated by
comparison with a microcystin LR standard curve. Quantitation with the colorimetric protein
phosphatase inhibition assay is based on a comparison with a microcystin LR standard curve. ELISA and
phosphatase assay results agree over a concentration range of 0.5 to 35 ug/g. Neither assay is specific for
a particular isoform.
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Please note: These procedures have been applied to analysis of algae products. Further research is
needed to develop and standardize the protocols for other sample types.
Source: Lawrence, J.F., Niedzwiadek, B., Menard, C., Lau, B.P., Lewis, D., Kuper-Goodman, T.,
Carbone, S., and Holmes, C. 2001. "Comparison of Liquid Chromatography/Mass Spectrometry, ELISA,
and Phosphatase Assay for the Determination of Microcystins in Blue-green Algae Products." Journal of
AOAC International 84(4): 1035-1044.
8.2.21 Literature Reference for Microcystins (Analyst. 1994. 119(7): 1525-1530)
These procedures should be used for confirmatory analysis of microcystins in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Microcystins
(Principal Isoforms: LR, YR, RR, LW)
Agent Category
Small Molecule
CASRN
77238-39-2
Procedures are discussed to test the presence of microcystin-LR, -LY, -LW, -LF, and -RR in treated and
untreated water samples. Cyanobacterial cells are separated from the water by filtration through 110-mm
GF/C discs. The cellular components collected on the discs are extracted three times with methanol; the
collected extraction fluids are combined and dried. The residue is resuspended in methanol and analyzed
by photodiode-array high-performance liquid chromatography (PDA-HPLC). The liquid portion of the
filtered water sample is subjected to trace enrichment using a Qg solid-phase extraction cartridge,
followed by identification and determination by PDA-HPLC. This procedure can detect microcystin
concentrations as low as 250 ng/L and is the basis of the World Health Organization (WHO) method for
the detection of microcystins.
Please note: These procedures have been applied to analysis of water. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Lawton, L.A., Edwards, C., and Codd, G.A. 1994. "Extraction and High-performance Liquid
Chromatographic Method for the Determination of Microcystins in Raw and Untreated Waters. "Analyst.
119(7): 1525-1530.
8.2.22 Literature Reference for Picrotoxin (Journal of Pharmaceutical & Biomedical
Analysis. 1989. 7(3): 369-375)
These procedures should be used for confirmatory analysis of picrotoxin in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Picrotoxin
Agent Category
Small Molecule
CASRN
124-87-8
Procedures are described for quantification of the two components of picrotoxin (picrotin and
picrotoxinin) in serum samples. Serum samples are prepared by washing with «-hexane, followed by
extraction with chloroform. The chloroform is evaporated and the sample is reconstituted in acetonitrile-
1 mM ammonium acetate buffer (pH 6.4) 34:66 (v/v) for assay by reversed-phase high-performance
liquid chromatography (HPLC). The effluent is monitored at 200 nm, and quantification is based on
peak-height ratio of analyte to the internal standard. A linear response is obtained for both analytes
(picrotin and picrotoxinin) in the range 0.2 to 20 (ig/mL.
SAM Revision 3.1
150
November 15, 2007
-------
Section 8 - Selected Biotoxin Methods
Please note: These procedures have been applied to analysis of serum. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Soto-Otero, R., Mendez-Alvarez, E., Sierra-Paredes, G., Galan-Valiente, J., Aguilar-Veiga, E.,
and Sierra-Marcuno, G. 1989. "Simultaneous Determination of the Two Components of Picrotoxin in
Serum by Reversed-phase High-performance Liquid Chromatography with Application to a
Pharmacokinetic Study in Rats." Journal of Pharmaceutical & Biomedical Analysis. 7(3): 369—375.
8.2.23 Literature Reference for Saxitoxin (Journal of AOAC International. 1995. 78(2):
528-532)
These procedures should be used for confirmatory analysis of saxitoxin in aerosol, solid, particulate,
liquid, and water samples.
Analyte(s)
Saxitoxin (SIX, NEOSAX,
GTX, dcGTX, dcSTX, SIX)
Agent Category
Small Molecule
CASRN
35523-89-8
Procedures are described to detect multiple analogues of saxitoxin in shellfish using ion-interaction
chromatography on a silica-based reversed-phase (C8) column with postcolumn periodate oxidation and
fluorescence detection. Toxin groups of different net charges are determined separately by isocratic
elution using either sodium 1-heptanesulfonate in ammonium phosphate (GTX-1, GTX-6, dcGTX2,
dcGTX3) or sodium 1-heplanesulfonate in ammonium phosphate and acetonitrile (STX, neoSTX,
dcSTX). For biological matrices, a cleanup procedure using a Ci8 solid-phase extraction cartridge is
effective in preventing false peaks. High sensitivity with detection limits ranging from 20 to 110 fmol are
achieved as a result of reduced band broadening and optimized reaction conditions. This method, when
applied to low-toxicity shellfish, gives higher values than the standard mouse bioassay.
Please note: These procedures have been applied to analysis of shellfish. Further research is needed to
develop and standardize the protocols for other sample types.
Source: Oshima, Y. 1995. "Postcolumn Derivatization Liquid Chromatographic Method for Paralytic
Shellfish Toxins." Journal of AOAC International. 78(2): 528-532.
8.2.24 Literature Reference for Shiga and Shiga-like Toxin (Journal of Clinical
Microbiology. 2007. 45(10): 3377-3380)
These procedures should be used for presumptive analysis of Shiga and Shiga-like toxins in aerosol,
solid, particulate, liquid, and water samples.
Analyte(s)
Shiga and Shiga-like Toxins
(Stx, Stx-1 , Stx-2)
Agent Category
Protein
CASRN
75757-64-1
Procedures are described for a rapid optical immunoassay for the detection of Shiga toxin Types 1 (Stx-1)
and 2 (Stx-2) using a commercially available kit. Fecal samples (742 specimens) are assayed for Shiga
toxins with and without enrichment of the specimens in broth. Duplicate assays are applied using either
the rapid optical immunoassay or a commercially available ELISA kit. Samples producing positive
results by immunoassay are confirmed by Vero cell cytotoxicity assay. Sensitivities of 96.8% are
achieved for direct stool sample assays.
Please note: These procedures have not been applied to analysis of environmental samples. Further
research is needed to develop and standardize the protocols for these sample types.
SAM Revision 3.1 151 November 15, 2007
-------
Section 8 - Selected Biotoxin Methods
Source: Teel, L.D., Daly, J.A., Jerris, R.C., Maul, D., Svanas, G., O'Brien, A.D., Park, C.H. "Rapid
Detection of Shiga Toxin-Producing Escherichia coll by Optical Immunoassay." Journal of Clinical
Microbiology. 45(10): 3377-3380.
8.2.25 Literature Reference for Tetrodotoxin (Journal of Clinical Laboratory Analysis.
1992. 6(2): 65-72)
These procedures should be used for presumptive analysis of tetrodotoxin (TTX) in aerosol, solid,
)articulate, liquid, and water samples.
Analyte(s)
Tetrodotoxin
Agent Category
Small Molecule
CASRN
9014-39-5
Procedures are described for a competitive inhibition enzyme immunoassay (CIEIA) for TTX in
biological samples. An anti-TTX monoclonal antibody (Mab), designated T20G10, is directly labeled
with alkaline phosphatase for use in the assay. Sensitivities of 6 to 7 ng/mL (1C 50) and 2 to 3 ng/mL
(IC20) are achieved.
Please note: These procedures have been applied to analysis of biological samples. Further research is
needed to develop and standardize the protocols for other sample types.
Source: Raybould, T.J., Bignami, G.S., Inouye, L.K., Simpson, S.B., Byrnes, J.B., Grothaus, P.G., and
Vann, D.C. 1992. "A Monoclonal Antibody-based Immunoassay for Detecting Tetrodotoxin in
Biological Samples." Journalof Clinical Laboratory Analysis. 6(2): 65—72.
8.2.26 Literature Reference for Tetrodotoxin (Analytical Biochemistry. 2001. 290(1): 10-
17)
These procedures should be used for confirmatory analysis of tetrodotoxin (TTX) in aerosol, solid,
)articulate, liquid, and water samples.
Analyte(s)
Tetrodotoxin
Agent Category
Small Molecule
CASRN
9014-39-5
Procedures are described for liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-
MS) analysis of TTXs in tissue samples from puffer fish and newts by a combination of chromatography
on a reversed-phase column with long carbon chains (C30) and with the mobile phase containing an ion
pair reagent (ammonium heptafluorobutyrate). The relationship between the amount of applied standard
TTX and its peak area on the mass chromatogram (m/z 320) shows good linearity over a range of 50 to
1000 pmol. The detection limit of TTX in the selected ion monitoring (SIM) mode is estimated to be 0.7
pmol, with a signal to noise ratio of 2:1.
Please note: These procedures have been applied to analysis of tissue samples. Further research is
needed to develop and standardize the protocols for other sample types.
Source: Shoji, Y., Yotsu-Yamashita, M., Miyazawa, T., and Yasumoto, T. 2001. "Electrospray
Ionization Mass Spectrometry of Tetrodotoxin and its Analogs: Liquid Chromatography/Mass
Spectrometry, Tandem Mass Spectrometry, and Liquid Chromatography/Tandem Mass Spectrometry."
Analytical Biochemistry. 290(1): 10-17.
SAM Revision 3.1 152 November 15, 2007
-------
Section 8 - Selected Biotoxin Methods
8.2.27 Lateral Flow Immunoassay Kits
These procedures should be used for presumptive analysis of botulinum neurotoxins Types A and B and
ricin in aerosol samples.
Analyte(s)
Botulinum neurotoxins
(Types A, B)
Ricin
Agent Category
Protein
Protein
CASRN
NA
9009-86-3
Test strips are self-contained, qualitative assays for screening environmental samples for the presence of
botulinum toxin and ricin. After the sample is collected, it is transferred onto the test strip where dye-
labeled antibodies detect trace amounts of the contaminant, as indicated by the presence of two bands in
the test result window. After 15 minutes, the results are read visually. Botulinum neurotoxin Type A can
be detected at 5 mg/L and Type B at 4 mg/L, 33% of the time. Ricin toxin can be detected at 20 mg/L,
with no cross-reactivity certain substances (i.e., lectin from soybeans).
An alternative lateral flow immunochromatographic device also can be used. This device uses two
antibodies in combination to specifically detect target antigen in solution. When a sufficient amount of
target antigen is present, the colloidal gold label accumulates in the sample window on a test strip,
forming a visible reddish-brown colored line. The presence of two bands indicates a positive reading.
Botulinum neurotoxin Type A can be detected at 0.01 mg/L and Type B at 0.5 mg/L, with no false
negatives detected when interferents are present. Ricin toxin can be detected at 0.035 mg/L, with 88%
accuracy.
These two lateral flow immunoassay kits have been evaluated by the U.S. EPA Environmental
Technology Verification (ETV) Program (http://wmv.epa.gOv/etv/pdfs/vrvs/0 l_vr_badd.pdf and
http://www.epa.gov/etv/pdfs/vrvs/01_vr_biothreat.pdf) for the detection of botulinum neurotoxins Types
A and B and ricin. Information regarding the evaluation of test strips can be accessed at these sites.
Please note: These procedures have been applied to analysis of toxin in buffer or water. Further research
is needed to develop and standardize the protocols for other sample types.
Source: Environmental Technology Verification. 2006. httpV/www.epa.gQY/ety/
SAM Revision 3.1 153 November 15, 2007
-------
Section 8 - Selected Biotoxin Methods
SAM Revision 3.1 154 November 15, 2007
-------
Section 9 - Conclusions
Section 9.0: Conclusions
Methods listed in Appendix A (chemical methods), Appendix B (radiochemical methods), Appendix C
(pathogen methods), and Appendix D (biotoxin methods) are recommended for use in assessment of the
extent of contamination and the effectiveness of decontamination following a homeland security event.
The primary objective of this document is not necessarily to identify the "best" analytical methods, but
rather to identify appropriate methods that represent a balance between providing existing, documented
determinative techniques and providing consistent and valid analytical results. The method selected for
each analyte/sample type combination was deemed the most general, appropriate, and broadly applicable
of available methods. This is a living guidance document for use by EPA and EPA-contracted
laboratories tasked with analysis of environmental samples following a homeland security event.
Recommended methods are subject to change based on procedure testing and advances in technology.
Any questions concerning the information in this document should be directed to the appropriate point(s)
of contact listed in Section 4.
SAM Revision 3.1 155 November 15, 2007
-------
Section 9 - Conclusions
SAM Revision 3.1 156 November 15, 2007
-------
Appendix A - Selected Chemical Methods
Appendix A: Selected Chemical Methods
SAM Revision 3.1 November 15, 2007
-------
Appendix A - Selected Chemical Methods
SAM Revision 3.1 November 15, 2007
-------
Appendix A: Selected Chemical Methods
Analyte(s)
Acrylamide
Acrylonitrile
Aldicarb (Temik)
Allyl alcohol
2-Amino-4,6-dinitrotoluene
(2-Am-DNT)
4-Amino-2,6-dinitrotoluene
(4-Am-DNT)
4-Aminopyridine
Ammonia
Ammonium metavanadate
(analyze for total vanadium)
Arsenic, Total
Arsenic trioxide
(analyze for total arsenic)
CASRN
79-06-1
107-13-1
116-06-3
107-18-6
35572-78-2
19406-51-0
504-24-5
7664-41-7
7803-55-6
7440-38-2
1327-53-3
Determinative
Technique
HPLC
HPLC
HPLC
GC-MS
HPLC
HPLC
HPLC
Spectrophotometry
ICP-MS / ICP-AES
ICP-MS / ICP-AES
ICP-MS / ICP-AES
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
Water extraction
8316
(EPASW-846)
Water extraction
8316
(EPASW-846)
831 8A
(EPASW-846)
5035A
(EPASW-846)
8260C
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
Not of concern
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
Water extraction
8316
(EPASW-846)
Water extraction
8316
(EPASW-846)
831 8A
(EPASW-846)
3585
(EPASW-846)
8260C
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
Not of concern
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
Aqueous Liquid
Samples
8316
(EPASW-846)
8316
(EPASW-846)
831 8A
(EPASW-846)
5030C
(EPASW-846)
8260C
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
4500- NH3 B
(SM)
4500- NH3G
(SM)
200.8
(EPA OW)
200.8
(EPA OW)
200.8
(EPA OW)
Drinking Water
Samples
8316
(EPASW-846)
8316
(EPASW-846)
531.2
(EPAOW)
5030C
(EPASW-846)
8260C
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
350.1
(EPAOW)
200.8
(EPAOW)
200.8
(EPAOW)
200.8
(EPAOW)
Air Samples
PV2004
(OS HA)
PV2004
(OS HA)
5601
(NIOSH)
TO-152
(EPAORD)
Not of concern
Not of concern
Not of concern
6015
(NIOSH)
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
SAM Revision 3.1, Appendix A
A-1
November 15, 2007
-------
Analyte(s)
Arsine
Asbestos
Boron trifluoride
Brodifacoum
Bromadiolone
Carbofuran (Furadan)
Carbon disulfide
Chlorfenvinphos
Chlorine
2-Chloroethanol
3-Chloro-1 ,2-propanediol
CASRN
7784-42-1
1332-21-4
7637-07-2
56073-10-0
28772-56-7
1563-66-2
75-15-0
470-90-6
7782-50-5
107-07-3
96-24-2
Determinative
Technique
GFAA
TEM
ISE
HPLC
HPLC
HPLC
GC-MS
GC-MS
Spectrophotometry
GC-MS /GC-FID
GC-MS
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
3050B
(EPASW-846)
7010
(EPASW-846)
D5755-03 (soft surfaces-
microvac) or D6480-99
(hard surfaces-wipes)
(ASTM)
Not of concern
3545A/3541
(EPASW-846)
8321 B
(EPASW-846)
3545A/3541
(EPASW-846)
8321 B
(EPASW-846)
831 8A
(EPASW-846)
5035A
(EPASW-846)
8260C
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
Not of concern
5035A
(EPASW-846)
8260C
(EPA SW-846)
3545A/3541
(EPASW-846)
8270D3
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
Not of concern
Not of concern
Not of concern
3580A
(EPASW-846)
8321 B
(EPASW-846)
3580A
(EPASW-846)
8321 B
(EPASW-846)
831 8A
(EPASW-846)
3585
(EPASW-846)
8260C
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
3585
(EPASW-846)
8260C
(EPA SW-846)
3580A
(EPASW-846)
8270D3
(EPASW-846)
Aqueous Liquid
Samples
200.8
(EPA OW)
Not of concern
Not of concern
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
831 8A
(EPASW-846)
5030C
(EPASW-846)
8260C
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
4500-CI G
(SM)
5030C
(EPASW-846)
8260C
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D3
(EPASW-846)
Drinking Water
Samples
200.8
(EPAOW)
Not of concern
Not of concern
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
531.2
(EPAOW)
524.2
(EPAOW)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
4500-CI G
(SM)
5030C
(EPASW-846)
8260C
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D3
(EPASW-846)
Air Samples
6001
(NIOSH)
10312:1995
(ISO)
ID-216SG
(OS HA)
Not of concern
Not of concern
5601
(NIOSH)
TO-15
(EPAORD)
TO-10A
(EPAORD)
Analyst, Vol. 124,
1999, pp. 1853-1857
4500-CI G
(SM)
2513
(NIOSH)
TO-10A4
(EPAORD)
SAM Revision 3.1, Appendix A
A-2
November 15, 2007
-------
Analyte(s)
Chloropicrin
Chlorosarin
Chlorosoman
2-Chlorovinylarsonous acid
(CVAA) (degradation product of
Lewisite)
Chlorpyrifos
Crimidine
Cyanide, Total
Cyanogen chloride
Cyclohexyl sarin (GF)
1 ,2-Dichloroethane
(degradation product of HD)
Dichlorvos
CASRN
76-06-2
1445-76-7
7040-57-5
85090-33-1
2921-88-2
535-89-7
57-12-5
506-77-4
329-99-7
107-06-2
62-73-7
Determinative
Technique
GC-MS
GC-MS
GC-MS
ICP-MS / ICP-AES
GC-MS
GC-MS
Spectrophotometry
GC-MS
GC-MS
GC-MS
GC-MS
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
3545A
(EPASW-846)
8270D5
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3050B
(EPA SW-846)
6020A/6010C
(EPA SW-846)
3545A/3541
(EPA SW-846)
8270D
(EPA SW-846)
3545A
(EPA SW-846)
8270D6
(EPASW-846)
ILM05.3CN
(EPA CLP)
5035A
(EPASW-846)
8260C
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPA SW-846)
5035A
(EPA SW-846)
8260C
(EPA SW-846)
3545A
(EPA SW-846)
8270D
(EPA SW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
3580A
(EPASW-846)
8270D5
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3031
(EPA SW-846)
6020A/6010C
(EPA SW-846)
3580A
(EPA SW-846)
8270D
(EPA SW-846)
3580A
(EPA SW-846)
8270D6
(EPASW-846)
Not of concern
3585
(EPASW-846)
8260C
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPA SW-846)
3585
(EPA SW-846)
8260C
(EPA SW-846)
3580A
(EPA SW-846)
8270D
(EPA SW-846)
Aqueous Liquid
Samples
551.1
(EPA OW)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
200.8
(EPA OW)
3520C/3535A
(EPA SW-846)
8270D
(EPA SW-846)
3535A
(EPA SW-846)
8270D6
(EPASW-846)
ILM05.3CN
(EPA CLP)
5030C
(EPASW-846)
8260C
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
5030C
(EPA SW-846)
8260C
(EPA SW-846)
3535A
(EPA SW-846)
8270D
(EPA SW-846)
Drinking Water
Samples
551.1
(EPA OW)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
200.8
(EPAOW)
3520C/3535A
(EPA SW-846)
8270D
(EPA SW-846)
3535A
(EPA SW-846)
8270D6
(EPASW-846)
335.4
(EPAOW)
5030C
(EPASW-846)
8260C
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
524.2
(EPA OW)
525.2
(EPA OW)
Air Samples
PV2103(OSHA)
TO-10A4
(EPAORD)
TO-10A4
(EPAORD)
IO-3.1
(EPA ORD)
IO-3.4/IO-3.5
(EPA ORD)
TO-10A
(EPA ORD)
Not of concern
6010
(NIOSH)
TO-15
(EPAORD)
TO-10A
(EPAORD)
TO-15
(EPAORD)
TO-10A
(EPA ORD)
SAM Revision 3.1, Appendix A
A-3
November 15, 2007
-------
Analyte(s)
Dicrotophos
Diesel Range Organics
Diisopropyl methylphosphonate
(DIMP)
(degradation product of GB)
Dimethylphosphite
Dimethylphosphoramidic acid
(degradation product of GA)
3,5-Dinitroaniline (3,5-DNA)
1,3-Dinitrobenzene (1,3-DNB)
2,4-Dinitrotoluene (2,4-DNT)
2,6-Dinitrotoluene (2,6-DNT)
Diphacinone
1,4-Dithiane
(degradation product of HD)
CASRN
141-66-2
NA
1445-75-6
868-85-9
33876-51-6
618-87-1
99-65-0
121-14-2
606-20-2
82-66-6
505-29-3
Determinative
Technique
GC-MS
GC-FID
GC-MS
GC-MS
GC-MS
HPLC
HPLC
HPLC
HPLC
HPLC
GC-MS
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
3545A
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPASW-846)
801 5C
(EPASW-846)
3545A
(EPASW-846)
8270D
(EPASW-846)
3545A
(EPASW-846)
8270D
(EPASW-846)
3545A
(EPASW-846)
832 1B7
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
3545A/3541
(EPASW-846)
8321 B
(EPASW-846)
3545A
(EPASW-846)
8270D9
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
801 5C
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
832 1B7
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
3580A
(EPASW-846)
8321 B
(EPASW-846)
3580A
(EPASW-846)
8270D9
(EPASW-846)
Aqueous Liquid
Samples
3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
801 5C
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
832 1B7
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
3535A
(EPASW-846)
8270D9
(EPASW-846)
Drinking Water
Samples
3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
801 5C
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
832 1B7
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
3535A
(EPASW-846)
8270D9
(EPASW-846)
Air Samples
TO-10A
(EPAORD)
Not of concern
TO-10A4
(EPAORD)
TO-10A
(EPAORD)
TO-10A8
(EPAORD)
Not of concern
Not of concern
Not of concern
Not of concern
Not of concern
Not of concern
SAM Revision 3.1, Appendix A
A-4
November 15, 2007
-------
Analyte(s)
EA2192 [Diisopropylaminoethyl
methylthiolophosphonate]
(hydrolysis product of VX)
Ethyl methylphosphonic acid
(EMPA)
(degradation product of VX)
Ethyldichloroarsine (ED)
N-Ethyldiethanolamine (EDEA)
(degradation product of HN-1)
Ethylene oxide
Fenamiphos
Fentanyl
Fluoride
Fluoroacetic acid and fluoroacetate
salts (analyze for fluoroacetate ion)
Formaldehyde
Gasoline Range Organics
CASRN
73207-98-4
1832-53-7
598-14-1
139-87-7
75-21-8
22224-92-6
437-38-7
16984-48-8
NA
50-00-0
NA
Determinative
Technique
GC-MS
GC-MS
GC-MS
GC-MS
GC-MS
GC-MS
HPLC
1C
1C
HPLC
GC-FID
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
3545A
(EPASW-846)
832 1B7
(EPASW-846)
3545A
(EPASW-846)
832 1B7
(EPASW-846)
3545A
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPASW-846)
8321 B
(EPASW-846)
5035A
(EPASW-846)
8260C
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPA SW-846)
8321 B
(EPA SW-846)
Not of concern
Analytical Letters, 1994,
27(14):
2703-2718
300.1, Rev 1.0
(EPA OW)
831 5A
(EPA SW-846)
5035A
(EPASW-846)
801 5C
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
3580A
(EPASW-846)
832 1B7
(EPASW-846)
3580A
(EPASW-846)
832 1B7
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8321 B
(EPASW-846)
3585
(EPASW-846)
8260C
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8321 B
(EPASW-846)
Not of concern
Analytical Letters,
1994,27(14):
2703-2718
300.1, Rev 1.0
(EPAOW)
Not of concern
3585
(EPASW-846)
801 5C
(EPASW-846)
Aqueous Liquid
Samples
3535A
(EPASW-846)
832 1B7
(EPASW-846)
3535A
(EPASW-846)
832 1B7
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
5030C
(EPASW-846)
8260C
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
300.1, Rev 1.0
(EPA OW)
300.1, Rev 1.0
(EPA OW)
831 5A
(EPASW-846)
5030C
(EPASW-846)
801 5C
(EPASW-846)
Drinking Water
Samples
3535A
(EPASW-846)
832 1B7
(EPASW-846)
3535A
(EPASW-846)
832 1B7
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
5030C
(EPASW-846)
8260C
(EPASW-846)
525.2
(EPAOW)
3520C/3535A
(EPA SW-846)
8321 B
(EPASW-846)
300.1, Rev 1.0
(EPA OW)
300.1, Rev 1.0
(EPA OW)
831 5A
(EPASW-846)
5030C
(EPASW-846)
801 5C
(EPASW-846)
Air Samples
TO-10A8
(EPAORD)
TO-10A8
(EPAORD)
TO-15
(EPAORD)
TO-10A
(EPAORD)
TO-15
(EPAORD)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
Not of concern
S301-1
(NIOSH)
300.1, Rev 1.0
(EPAOW)
2016
(NIOSH)
Not of concern
SAM Revision 3.1, Appendix A
A-5
November 15, 2007
-------
Analyte(s)
Hexahydro-1 ,3,5-trinitro-1 ,3,5-
triazine (RDX)
Hexamethylenetriperoxidediamine
(HMTD)
Hydrogen bromide
Hydrogen chloride
Hydrogen cyanide
Hydrogen fluoride
Hydrogen sulfide
Isopropyl methylphosphonic acid
(IMPA) (degradation product of
GB)
Kerosene
Lewisite 1 (L-1)11
[2-chlorovinyldichloroarsine]
(analyze for total arsenic)
Lewisite 2 (L-2)
[bis(2-chlorovinyl)chloroarsine]
(analyze for total arsenic)
Lewisite 3 (L-3)
[tris(2-chlorovinyl)arsine]
(analyze for total arsenic)
CASRN
121-82-4
283-66-9
10035-10-6
7647-01-0
74-90-8
7664-39-3
7783-06-4
1832-54-8
64742-81-0
541-25-3
40334-69-8
40334-70-1
Determinative
Technique
HPLC
HPLC
1C
1C
Spectrophotometry
1C
1C
GC-MS
GC-FID
ICP-MS / ICP-AES
ICP-MS / ICP-AES
ICP-MS / ICP-AES
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
8330B
(EPASW-846)
8330B
(EPASW-846)
Not of concern
Not of concern
Not of concern
Not of concern
Not of concern
3545A
(EPASW-846)
832 1B7
(EPASW-846)
5035A
(EPASW-846)
801 5C
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPA SW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
8330B
(EPASW-846)
8330B
(EPASW-846)
Not of concern
Not of concern
Not of concern
Not of concern
Not of concern
3580A
(EPASW-846)
832 1B7
(EPASW-846)
3585
(EPASW-846)
801 5C
(EPASW-846)
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
3031
(EPASW-846)
6020A/6010C
(EPA SW-846)
Aqueous Liquid
Samples
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
Not of concern
Not of concern
ILM05.3CN
(EPA CLP)
Not of concern
Not of concern
3535A
(EPASW-846)
832 1B7
(EPASW-846)
5030C
(EPASW-846)
801 5C
(EPASW-846)
200.8
(EPA OW)
200.8
(EPA OW)
200.8
(EPA OW)
Drinking Water
Samples
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
Not of concern
Not of concern
335.4
(EPAOW)
Not of concern
Not of concern
3535A
(EPASW-846)
832 1B7
(EPASW-846)
5030C
(EPASW-846)
801 5C
(EPASW-846)
200.8
(EPAOW)
200.8
(EPAOW)
200.8
(EPAOW)
Air Samples
Not of concern
Not of concern
7903
(NIOSH)
7903
(NIOSH)
6010
(NIOSH)
790310
(NIOSH)
6013
(NIOSH)
TO-10A8
(EPAORD)
Not of concern
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
SAM Revision 3.1, Appendix A
A-6
November 15, 2007
-------
Analyte(s)
Lewisite oxide
(degradation product of Lewisite)
Mercury, Total
Methamidophos
Methomyl
Methoxyethylmercuric acetate
(analyze for total mercury)
Methyl acrylonitrile
Methyl fluoroacetate
(analyze for fluoroacetate ion)
Methyl hydrazine
Methyl isocyanate
Methyl parathion
Methyl-2,4,6-trinitrophenylnitramine
(Tetryl)
CASRN
1306-02-1
7439-97-6
10265-92-6
16752-77-5
151-38-2
126-98-7
453-18-9
60-34-4
624-83-9
298-00-0
479-45-8
Determinative
Technique
ICP-MS / ICP-AES
Thermal
Decomposition AA /
CVAA / CVAFS
GC-MS
HPLC
Thermal
Decomposition AA /
CVAA / CVAFS
HPLC-UV
1C
GC-MS /
Spectrophotometry
HPLC
GC-MS
HPLC
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
747312
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
831 8A
(EPASW-846)
747312
(EPASW-846)
Water extraction
8316
(EPASW-846)
Analytical Letters, 1994,
27 (14): 2703-2718
300.1, Rev 1.0
(EPA OW)
3545A/3541
(EPA SW-846)
8270D
(EPA SW-846)
Not of concern
3545A
(EPA SW-846)
8270D
(EPA SW-846)
8330B
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
Not of concern
3580A
(EPASW-846)
8270D
(EPASW-846)
831 8A
(EPASW-846)
Not of concern
Water extraction
8316
(EPASW-846)
Analytical Letters,
1994, 27 (14): 2703-
2718
300.1, Rev 1.0
(EPAOW)
3580A
(EPASW-846)
8270D
(EPA SW-846)
Not of concern
3580A
(EPA SW-846)
8270D
(EPA SW-846)
8330B
(EPASW-846)
Aqueous Liquid
Samples
200.8
(EPA OW)
747312
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
831 8A
(EPASW-846)
747312
(EPASW-846)
8316
(EPASW-846)
300.1, Rev 1.0
(EPA OW)
3520C/3535A
(EPA SW-846)
8270D
(EPA SW-846)
Not of concern
3535A
(EPA SW-846)
8270D
(EPA SW-846)
3535A/8330B
(EPA SW-846)
8330B
(EPASW-846)
Drinking Water
Samples
200.8
(EPAOW)
245.2
(EPAOW)
3535A
(EPASW-846)
8270D
(EPASW-846)
531.2
(EPAOW)
245.2
(EPAOW)
8316
(EPASW-846)
300.1, Rev 1.0
(EPAOW)
3520C/3535A
(EPA SW-846)
8270D
(EPA SW-846)
Not of concern
3535A
(EPA SW-846)
8270D
(EPA SW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
Air Samples
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
IO-5
(EPAORD)
TO-10A13(EPAORD)
5601
(NIOSH)
IO-5
(EPAORD)
PV2004
(OS HA)
S301-1
(NIOSH)
300.1, Rev 1.0
(EPAOW)
3510
(NIOSH)
OS HA 54
TO-10A
(EPAORD)
Not of concern
SAM Revision 3.1, Appendix A
A-7
November 15, 2007
-------
Analyte(s)
Methylamine
N-Methyldiethanolamine (MDEA)
(degradation product of HN-2)
1-Methylethyl ester
ethylphosphonofluoridic acid (GE)
Methylphosphonic acid (MPA)
(degradation product of VX, GB, &
GD)
Mevinphos
Mustard, nitrogen (HN-1)
[bis(2-chloroethyl)ethylamine]
Mustard, nitrogen (HN-2)
[2,2'-dichloro-N-methyldiethylamine
N,N-bis(2-chloroethyl)methylamine]
Mustard, nitrogen (HN-3)
[tris(2-chloroethyl)amine]
Mustard, sulfur / Mustard gas (HD)
Nicotine sulfate
Nitrobenzene (NB)
CASRN
74-89-5
105-59-9
1189-87-3
993-13-5
7786-34-7
538-07-8
51-75-2
555-77-1
505-60-2
54-11-5
98-95-3
Determinative
Technique
HPLC
GC-MS
GC-MS
GC-MS
GC-MS
GC-MS
GC-MS
GC-MS
GC-MS
GC-MS
HPLC
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
Not of concern
3545A/3541
(EPASW-846)
8321 B
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3545A
(EPASW-846)
832 1B7
(EPASW-846)
3545A
(SW-846)
8270D
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPA SW-846)
3545A/3541
(EPA SW-846)
8270D
(EPA SW-846)
3571
(EPA SW-846)
8270D14
(EPASW-846)
3545A
(EPASW-846)
8270D
(EPASW-846)
8330B
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
Not of concern
3580A
(EPASW-846)
8321 B
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
832 1B7
(EPASW-846)
3580A
(SW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPA SW-846)
3580A
(EPA SW-846)
8270D
(EPA SW-846)
3571
(EPA SW-846)
8270D14
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
8330B
(EPASW-846)
Aqueous Liquid
Samples
Not of concern
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
832 1B7
(EPASW-846)
3535A
(SW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
3520C/3535A
(EPA SW-846)
8270D
(EPA SW-846)
3571
(EPA SW-846)
8270D14
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
Drinking Water
Samples
Not of concern
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
832 1B7
(EPASW-846)
525.2 (OW)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
3520C/3535A
(EPA SW-846)
8270D
(EPA SW-846)
3571
(EPA SW-846)
8270D14
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
Air Samples
OS HA 40
TO-10A
(EPAORD)
TO-10A4
(EPAORD)
TO-10A8
(EPAORD)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
Not of concern
Not of concern
SAM Revision 3.1, Appendix A
A-8
November 15, 2007
-------
Analyte(s)
Nitroglycerin (NG)
2-Nitrotoluene (2-NT)
3-Nitrotoluene (3-NT)
4-Nitrotoluene (4-NT)
Octahydro-1 ,3,5,7-tetranitro-
1,3,5,7-tetrazocine(HMX)
Organophosphate pesticides, NOS
Osmium tetroxide
(analyze for total osmium)
Oxamyl
Paraquat
Parathion
Pentaerythritol tetranitrate (PETN)
CASRN
55-63-0
88-72-2
99-08-1
99-99-0
2691-41-0
NA
20816-12-0
23135-22-0
4685-14-7
56-38-2
78-11-5
Determinative
Technique
HPLC
HPLC
HPLC
HPLC
HPLC
GC-MS/GC-NPD/
GC-FPD
ICP-AES/GFAA
HPLC
HPLC
GC-MS
HPLC
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3050B
(EPASW-846)
601 OC
(EPASW-846)
831 8A
(EPASW-846)
Not of concern
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
8330B
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
831 8A
(EPASW-846)
Not of concern
3580A
(EPASW-846)
8270D
(EPASW-846)
8330B
(EPASW-846)
Aqueous Liquid
Samples
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
614
(EPA OW)
252.2
(EPA OW)
831 8A
(EPASW-846)
549.2
(EPA OW)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
Drinking Water
Samples
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
507
(EPAOW)
252.2
(EPAOW)
531.2
(EPAOW)
549.2
(EPAOW)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
Air Samples
Not of concern
Not of concern
Not of concern
Not of concern
Not of concern
5600
(NIOSH)
IO-3.1
(EPAORD)
IO-3.4
(EPAORD)
5601
(NIOSH)
Not of concern
TO-10A
(EPAORD)
Not of concern
SAM Revision 3.1, Appendix A
A-9
November 15, 2007
-------
Analyte(s)
Perfluoroisobutylene (PFIB)
Phencyclidine
Phenol
Phorate
Phosgene
Phosphamidon
Phosphine
Phosphorus trichloride
Pinacolyl methyl phosphonic acid
(PMPA)
(degradation product of GD)
Propylene oxide
R 33 (VR)
[methylphosphonothioic acid, S-[2-
(diethylamino)ethyl] O-2-
methylpropyl ester]
CASRN
382-21-8
77-10-1
108-95-2
298-02-2
75-44-5
13171-21-6
7803-51-2
7719-12-2
616-52-4
75-56-9
159939-87-4
Determinative
Technique
GC-MS/GC-NPD
GC-MS
GC-MS
GC-MS
GC-NPD
GC-MS
Spectrophotometry
Spectrophotometry
GC-MS
GC-MS / GC-FID
GC-MS
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
Not of concern
3545A
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3545A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
Not of concern
Not of concern
3545A
(EPA SW-846)
832 1B7
(EPASW-846)
5035A
(EPASW-846)
8260C
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPA SW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
Not of concern
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
3580A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
Not of concern
3580A
(EPASW-846)
832 1B7
(EPASW-846)
3585
(EPASW-846)
8260C
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPA SW-846)
Aqueous Liquid
Samples
Not of concern
3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
Not of concern
Not of concern
3535A
(EPASW-846)
832 1B7
(EPASW-846)
5030C
(EPASW-846)
8260C
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
Drinking Water
Samples
Not of concern
3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
Not of concern
Not of concern
3535A
(EPASW-846)
832 1B7
(EPASW-846)
5030C
(EPASW-846)
8260C
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
Air Samples
OSHA6115
TO-10A
(EPA ORD)
TO-10A
(EPA ORD)
TO-10A
(EPA ORD)
OSHA 61
TO-10A
(EPA ORD)
6002
(NIOSH)
6402
(NIOSH)
TO-10A8
(EPA ORD)
1612
(NIOSH)
TO-10A
(EPA ORD)
SAM Revision 3.1, Appendix A
A-10
November 15, 2007
-------
Analyte(s)
Sarin (GB)
Semivolatile Organic Compounds,
NOS
Sodium arsenite
(analyze for total arsenic)
Sodium azide
(analyze for hydrazoic acid)
Soman (GD)
Strychnine
Sulfur dioxide
Sulfur trioxide
Tabun (GA)
Tear gas (CS) [chlorobenzylidene
malonitrile]
Tetraethyl pyrophosphate
CASRN
107-44-8
NA
7784-46-5
26628-22-8
96-64-0
57-24-9
7446-09-5
7446-11-9
77-81-6
2698-41-1
107-49-3
Determinative
Technique
GC-MS
GC-MS
ICP-MS / ICP-AES
1C
GC-MS
GC-MS
1C
Titrimetry
GC-MS
GC-MS
GC-MS
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
3571
(EPASW-846)
8270D14
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
J. of Forensic Science,
1998, 43(1): 200-20216
300.1, Revl.O17
(EPA OW)
3545A
(EPASW-846)
8270D
(EPASW-846)
3545A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
Not of concern
3545A
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3545A
(EPASW-846)
8270D
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
3571
(EPASW-846)
8270D14
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
3580A16
(EPASW-846)
300.1, Revl.O17
(EPAOW)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
Not of concern
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
Aqueous Liquid
Samples
3571
(EPASW-846)
8270D14
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
200.8
(EPA OW)
J. of Forensic Science,
1998, 43(1): 200-20216
300.1, Revl.O17
(EPA OW)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
Not of concern
3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
Drinking Water
Samples
3571
(EPASW-846)
8270D14
(EPASW-846)
525.2
(EPAOW)
200.8
(EPAOW)
J. of Forensic
Science, 1998,43(1):
200-20216
300.1, Revl.O17
(EPAOW)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
Not of concern
Not of concern
3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D
(EPASW-846)
Air Samples
TO-10A4
(EPAORD)
TO-10A
(EPAORD)
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
ID-211 (OHSA)
TO-10A4
(EPAORD)
Not of concern
6004
(NIOSH)
Method 8
(EPAOAQPS)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
SAM Revision 3.1, Appendix A
A-11
November 15, 2007
-------
Analyte(s)
Tetramethylenedisulfotetramine
Thallium sulfate
(analyze for total thallium)
Thiodiglycol (TDG)
(degradation product of HD)
1 ,4-Thioxane
(degradation product of HD)
Titanium tetrachloride (analyze for
total Titanium)
Triacetone triperoxide (TATP)
Triethanolamine (TEA)
(degradation product of HN-3)
Trimethyl phosphite
1 ,3,5-Trinitrobenzene (1,3,5-TNB)
2,4,6-Trinitrotoluene(2,4,6-TNT)
Vanadium pentoxide
(analyze for total vanadium)
CASRN
80-12-6
10031-59-1
111-48-8
15980-15-1
7550-45-0
17088-37-8
102-71-6
121-45-9
99-35-4
118-96-7
1314-62-1
Determinative
Technique
GC-MS
ICP-MS / ICP-AES
GC-MS
GC-MS
ICP-MS / ICP-AES
HPLC
GC-MS
GC-MS
HPLC
HPLC
ICP-MS / ICP-AES
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
3545A
(EPASW-846)
8270D
(EPASW-846)
3545A
(EPASW-846)
8270D9
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
8330B
(EPASW-846)
3545A/3541
(EPASW-846)
8321 B
(EPASW-846)
3545A
(EPASW-846)
8270D5
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
3050B
(EPASW-846)
6020A/6010C
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
3580A
(EPASW-846)
8270D
(EPASW-846)
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D9
(EPASW-846)
Not of concern
8330B
(EPASW-846)
3580A
(EPASW-846)
8321 B
(EPASW-846)
3580A
(EPASW-846)
8270D5
(EPASW-846)
8330B
(EPASW-846)
8330B
(EPASW-846)
3031
(EPASW-846)
6020A/6010C
(EPASW-846)
Aqueous Liquid
Samples
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
200.8
(EPA OW)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D9
(EPASW-846)
Not of concern
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
3535A
(EPASW-846)
8270D5
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
200.8
(EPA OW)
Drinking Water
Samples
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
200.8
(EPAOW)
3535A
(EPASW-846)
8270D
(EPASW-846)
3535A
(EPASW-846)
8270D9
(EPASW-846)
Not of concern
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3520C/3535A
(EPASW-846)
8321 B
(EPASW-846)
3535A
(EPASW-846)
8270D5
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
3535A/8330B
(EPASW-846)
8330B
(EPASW-846)
200.8
(EPAOW)
Air Samples
TO-10A
(EPAORD)
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
TO-10A
(EPAORD)
Not of concern
Not of concern
Not of concern
TO-10A
(EPAORD)
TO-10A
(EPAORD)
Not of concern
Not of concern
IO-3.1
(EPAORD)
IO-3.4/IO-3.5
(EPA ORD)
SAM Revision 3.1, Appendix A
A-12
November 15, 2007
-------
Analyte(s)
VE [phosphonothioic acid, ethyl-, S-
(2-(diethylamino)ethyl) O-ethyl
ester]
VG [phosphonothioic acid, S-(2-
(diethylamino)ethyl) O,O-diethyl
ester]
VM [phosphonothioic acid, methyl-,
S-(2-(diethylamino)ethyl) O-ethyl
ester]
VX [O-ethyl-S-(2-
diisopropylaminoethyl)methyl-
phosphonothiolate]
White phosphorus
CASRN
21738-25-0
78-53-5
21770-86-5
50782-69-9
12185-10-3
Determinative
Technique
GC-MS
GC-MS
GC-MS
GC-MS
GC-NPD / GC-FPD
Method Type
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Sample Prep
Determinative
Solid Samples
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPASW-846)
8270D
(EPASW-846)
3545A/3541
(EPA SW-846)
8270D
(EPA SW-846)
3571
(EPA SW-846)
8270D14
(EPASW-846)
7580
(EPASW-846)
Non-aqueous
Liquid/Organic Solid
Samples1
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPASW-846)
8270D
(EPASW-846)
3580A
(EPA SW-846)
8270D
(EPA SW-846)
3571
(EPA SW-846)
8270D14
(EPASW-846)
7580
(EPASW-846)
Aqueous Liquid
Samples
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
3520C/3535A
(EPA SW-846)
8270D
(EPA SW-846)
3571
(EPA SW-846)
8270D14
(EPASW-846)
7580
(EPASW-846)
Drinking Water
Samples
3520C/3535A
(EPASW-846)
8270D
(EPASW-846)
3520C/3535A
(EPASW-846)
8270D
(EPA SW-846)
3520C/3535A
(EPA SW-846)
8270D
(EPA SW-846)
3571
(EPA SW-846)
8270D14
(EPASW-846)
7580
(EPASW-846)
Air Samples
TO-10A
(EPAORD)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
TO-10A
(EPAORD)
7905
(NIOSH)
Footnotes
1 An organic solid sample is a solid that completely dissolves in an organic solvent and leaves no solid residue.
2 If problems occur when using this method, it is recommended that TO-10A be used.
3 For this analyte, SW-846 Method 8270D must be modified to include a derivatization step.
4 If problems occur when using this method, it is recommended that the canister Method TO-15 be used.
5 If problems occur with analyses, lower the injection temperature.
6 If problems occur when using this method, it is recommended that SW-846 Method 8321B be used as the determinative method. Sample preparation methods should remain the same.
7 LC-MS (electrospray) procedures are preferred for these analytes; however, if this technique is not available to the laboratory, GC-MS procedures using derivatization based on SW-846 Method 8270D may be
used. Sample preparation methods should remain the same. Both electrospray LC-MS and GC/MS derivatization procedures are currently under development.
8 For this analyte, Method TO-10A must be modified to include a derivatization step.
9 If problems occur when using this method, it is recommended that SW-846 Method 8260C and appropriate corresponding sample preparation procedures (i.e., 5035A for solid samples, 3585 for non-aqueous
liquid/organic solid samples, and 5030C for aqueous liquid and drinking water samples) be used.
10 If problems occur when using this method, it is recommended that NIOSH Method 7906 be used.
11 Laboratory testing is currently under way for speciation of Lewisite 1 using GC-MS techniques.
12 If equipment is not available, use CVAA Methods 7471B (EPA SW-846) for solid samples and 7470A (EPA SW-846) for aqueous liquid samples.
13 If problems occur when using this method, it is recommended that NIOSH Method 5600 be used.
14 For this analyte, refer to EPA SW-846 Method 8271 for GC-MS conditions.
15 If problems occur when using this method, it is recommended that a method based on the following journal article be used: J. Chrom. A, 1098: (2005) 156-165.
16 Water extraction, filtration, and acidification steps from the Journal of Forensic Science, 1998. 43(1):200-202 should be used for the preparation of solid samples. Filtration and acidification steps from this
journal should be used for preparation of aqueous liquid and drinking water samples. The acidification step from the journal should be used with EPA SW-846 Method 3580A for preparation of non-aqueous
liquid/organic solid samples.
17 If analyses are problematic, refer to column manufacturer for alternate conditions
SAM Revision 3.1, Appendix A
A-13
November 15, 2007
-------
SAM Revision 3.1, Appendix A A -14 November 15, 2007
-------
Appendix B - Selected Radiochemical Methods
Appendix B : Selected Radiochemical Methods
SAM Revision 3.1 November 15, 2007
-------
Appendix B - Selected Radiochemical Methods
SAM Revision 3.1 November 15, 2007
-------
Appendix B: Selected Radiochemical Methods
Analyte Class
Gross Alpha
Gross Beta
Gamma
Analyte(s)
Americium-241 2
Californium-2522
Cesium-1374
Cobalt-60
Curium-2442
Europium-154
lridium-192
Plutonium-2382
Plutonium-2392
Polonium-210
CASRN
14596-10-2
13981-17-4
10045-97-3
10198-40-0
13981-15-2
15585-10-1
14694-69-0
13981-16-3
15117-48-3
13981-52-7
Determinative
Technique
Alpha/Beta
spectrometry
Alpha/Beta
spectrometry
Gamma
spectrometry
Determinative
Technique
Alpha/Gamma
spectrometry
Alpha
spectrometry
Gamma
spectrometry
Gamma
spectrometry
Alpha
spectrometry
Gamma
spectrometry
Gamma
spectrometry
Alpha
spectrometry
Alpha
spectrometry
Alpha
spectrometry
Drinking Water Samples
900.0 (EPA)
900.0 (EPA)
901.1 (EPA)
Drinking Water Samples
Qualitative
Determination1
D3084
(ASTM)
D3084
(ASTM)
901.1
(EPA)
901.1
(EPA)
D3084
(ASTM)
901.1
(EPA)
901.1
(EPA)
D3084
(ASTM)
D3084
(ASTM)
Po-02-RC
(HASL-300)
Confirmatory
Am-04-RC
(HASL-300)
Am-04-RC
(HASL-300)
901.1
(EPA)
901.1
(EPA)
Am-04-RC
(HASL-300)
901.1
(EPA)
901.1
(EPA)
EMSL-33
(EPA)
EMSL-33
(EPA)
Po-02-RC
(HASL-300)
Aqueous and Liquid Phase
Samples
7110B(SM)
7110B(SM)
Ga-01-R
(HASL-300)
Aqueous and Liquid Phase
Samples
Qualitative
Determination1
D3084
(ASTM)
D3084
(ASTM)
7120
(SM)
7120
(SM)
D3084
(ASTM)
7120
(SM)
7120
(SM)
D3084
(ASTM)
D3084
(ASTM)
Po-02-RC
(HASL-300)
Confirmatory
Am-04-RC
(HASL-300)
Am-04-RC
(HASL-300)
7120
(SM)
7120
(SM)
Am-04-RC
(HASL-300)
7120
(SM)
7120
(SM)
EMSL-33
(EPA)
EMSL-33
(EPA)
Po-02-RC
(HASL-300)
Soil and Sediment Samples
AP1
(ORISE)
AP1
(ORISE)
Ga-01-R
(HASL-300)
Soil and Sediment Samples
Qualitative
Determination1
Am-02-RC
(HASL-300)
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
D3084
(ASTM)
D3084
(ASTM)
Po-02-RC
(HASL-300)
Confirmatory
Am-01-RC3
(HASL-300)
Am-01-RC3
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Am-01-RC3
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
EMSL-33
(EPA)
EMSL-33
(EPA)
Po-02-RC
(HASL-300)
Surface Wipes
FRMAC, Vol 2, pg. 33
FRMAC, Vol 2, pg. 33
Ga-01-R
(HASL-300)
Surface Wipes
Qualitative
Determination1
D3084
(ASTM)
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
D3084
(ASTM)
D3084
(ASTM)
Method 111
(EPA)
Confirmatory
Am-04-RC
(HASL-300)
Am-04-RC
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Am-04-RC
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
EMSL-33
(EPA)
EMSL-33
(EPA)
Method 111
(EPA)
Air Filters
FRMAC, Vol 2, pg. 33
FRMAC, Vol 2, pg. 33
Ga-01-R
(HASL-300)
Air Filters
Qualitative
Determination1
D3084
(ASTM)
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
D3084
(ASTM)
D3084
(ASTM)
Method 111
(EPA)
Confirmatory
Am-04-RC
(HASL-300)
Am-04-RC
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Am-04-RC
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
EMSL-33
(EPA)
EMSL-33
(EPA)
Method 111
(EPA)
SAM Revision 3.1, Appendix B
B-1
November 15, 2007
-------
Analyte(s)
Radium-2264
Ruthenium-103
Ruthenium-1064
Selenium-75
Strontium-904
Uranium-2382
CASRN
13982-63-3
13968-53-1
13967-48-1
14265-71-5
10098-97-2
7440-61-1
Determinative
Technique
Alpha Counting
Gamma
spectrometry
Gamma
spectrometry
Gamma
spectrometry
Beta counting by
low-background
gas flow
proportional
detector
Alpha Counting
Drinking Water Samples
Qualitative
Determination1
903.0
(EPA)
901.1
(EPA)
901.1
(EPA)
901.1
(EPA)
7500-Sr B
(SM)
908.0
(EPA)
Confirmatory
903.1
(EPA)
901.1
(EPA)
901.1
(EPA)
901.1
(EPA)
7500-Sr B
(SM)
D3972
(ASTM)
Aqueous and Liquid Phase
Samples
Qualitative
Determination1
7500-Ra B
(SM)
7120
(SM)
7120
(SM)
7120
(SM)
7500-Sr B
(SM)
7500-U B
(SM)
Confirmatory
7500-Ra C
(SM)
7120
(SM)
7120
(SM)
7120
(SM)
7500-Sr B
(SM)
7500-U C
(SM)
Soil and Sediment Samples
Qualitative
Determination1
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Sr-03-RC
(HASL-300)
D3084
(ASTM)
Confirmatory
EMSL-19
(EPA)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Sr-03-RC
(HASL-300)
EMSL-33
(EPA)
Surface Wipes
Qualitative
Determination1
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Sr-03-RC
(HASL-300)
D3084
(ASTM)
Confirmatory
EMSL-19
(EPA)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Sr-03-RC
(HASL-300)
EMSL-33
(EPA)
Air Filters
Qualitative
Determination1
D3084
(ASTM)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Sr-03-RC
(HASL-300)
D3084
(ASTM)
Confirmatory
EMSL-19
(EPA)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Ga-01-R
(HASL-300)
Sr-03-RC
(HASL-300)
EMSL-33
(EPA)
1 In those cases where the same method is listed for qualitative determination and confirmatory analysis, qualitative determination can be performed by application of the method over a shorter count time than
that used for confirmatory analysis.
2 If it is suspected that the sample exists in refractory form (i.e., non-digestable or dissolvable material after normal digestion methods) or if there is a matrix interference problem, use ORISE Method AP11 for
qualitative determination or confirmatory analysis.
3 In cases where only small sample volumes (£100 g) are available, use HASL-300 Method Pu-12-RC.
4 Methods identified will measure decay product of these isotopes.
SAM Revision 3.1, Appendix B
B-2
November 15, 2007
-------
Appendix C - Selected Pathosen Methods
Appendix C: Selected P athogen Methods
SAM Revision 3.1 November 15, 2007
-------
Appendix C - Selected Pathosen Methods
SAM Revision 3.1 November 15, 2007
-------
Appendix C: Pathogen Methods
Note: If viability determinations are needed (e.g., evaluation of the efficacy of disinfection), a viability-based procedure should be used. Those methods that include viability
procedures are indicated as culture, tissue culture, animal infectivity, or embryonation of eggs under the analytical technique column of this appendix, and usually include
biochemical and/or immunological differentiation of cultures.
Pathogen (s)
Viability
Analytical
Technique
Analytical
Method
Solid1
(soil, powder)
Particulate2
(swabs, wipes,
HEPA)
Liquid/Water
(filter, grab)
Drinking Water
(filter, grab)
Aerosol
(growth media,
filter, liquid)
Bacteria
Bacillus anthracis
[Anthrax]
Brucella spp.
(B. abortus, B. melitensis, B. suis )
Burkholderia mallei
[Glanders]
Burkholderia pseudomallei
[Melioidosis]
Campylobacterjejuni
Chlamydophila psittaci
(formerly known as Chlamydia psittaci)
Coxiella burnetii
[Q-fever]
Escherichia coli O157:H7
(E. coli)
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Culture
PCR/lmmunoassay
Culture
PCR/lmmunoassay
Culture
PCR/lmmunoassay
Culture
PCR/lmmunoassay
Culture
PCR
Tissue culture
PCR/lmmunoassay
Culture
PCR/lmmunoassay
Culture
Immunoassay
LRN
This pathogen is included in the HHS/USDA select agent list. Select agent methods are generally provided and controlled
by the LRN. In some cases, methods that apply to environmental samples may not be fully developed or validated.
If analysis of this agent is required, contact the LRN at (404) 639-2790 for information on the LRN laboratory capable of
receiving and processing the sample.
LRN
This pathogen is included in the HHS/USDA select agent list. Select agent methods are generally provided and controlled
by the LRN. In some cases, methods that apply to environmental samples may not be fully developed or validated.
If analysis of this agent is required, contact the LRN at (404) 639-2790 for information on the LRN laboratory capable of
receiving and processing the sample.
LRN
This pathogen is included in the HHS/USDA select agent list. Select agent methods are generally provided and controlled
by the LRN. In some cases, methods that apply to environmental samples may not be fully developed or validated.
If analysis of this agent is required, contact the LRN at (404) 639-2790 for information on the LRN laboratory capable of
receiving and processing the sample.
LRN
This pathogen is included in the HHS/USDA select agent list. Select agent methods are generally provided and controlled
by the LRN. In some cases, methods that apply to environmental samples may not be fully developed or validated.
If analysis of this agent is required, contact the LRN at (404) 639-2790 for information on the LRN laboratory capable of
receiving and processing the sample.
SM 9260 G
Molecular and
Cellular Probes 20:
269-279
Journal of Clinical
Microbiology
38: 1085-1093
Journal of Clinical
Microbiology
38: 1085-1093
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
SM 9260 G
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
SM 9260 G
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Unlikely to be viable
Unlikely to be viable
Adapted from
analytical method
Adapted from
analytical method
LRN
This pathogen is included in the HHS/USDA select agent list. Select agent methods are generally provided and controlled
by the LRN. In some cases, methods that apply to environmental samples may not be fully developed or validated.
If analysis of this agent is required, contact the LRN at (404) 639-2790 for information on the LRN laboratory capable of
receiving and processing the sample.
SM 9260 F
SM 9260 F
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
SM 9260 F
SM 9260 F
SM 9260 F
SM 9260 F
Unlikely to be viable
Unlikely to be viable
SAM Revision 3.1, Appendix C
C-1
November 15, 2007
-------
Pathogen (s)
Francisella tularensis
Uularemia]
Leptospira
(L. interrogans Serovars
Icteroheamorrhagiae, Autralis, Balum,
Bataviae, Bejro, Pomona)
Listeria monocytogenes
Salmonella spp.
(Method not applicable to Salmonella typhi)
Salmonella typhi
[Typhoid fever]
Shigella spp.
[Shigellosis]
Staphylococcus aureus
Vibrio cholerae O1 and O139
[Cholera]
Yersinia pestis
[Plague]
Viability
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Analytical
Technique
Culture
PCR/lmmunoassay
Culture
Immunoassay
Culture
Immunoassay
Culture
Immunoassay
Culture
Immunoassay
Culture
Immunoassay
Culture
TBD
Culture
Immunoassay
Culture
PCR/lmmunoassay
Analytical
Method
Solid1
(soil, powder)
Particulate2
(swabs, wipes,
HEPA)
Liquid/Water
(filter, grab)
Drinking Water
(filter, grab)
Aerosol
(growth media,
filter, liquid)
LRN
This pathogen is included in the HHS/USDA select agent list. Select agent methods are generally provided and controlled
by the LRN. In some cases, methods that apply to environmental samples may not be fully developed or validated.
If analysis of this agent is required, contact the LRN at (404) 639-2790 for information on the LRN laboratory capable of
receiving and processing the sample.
SM 9260 I
SM 9260 I
FDA/Bacteriological
Analytical Manual
Chapter 10, 2003
FDA/Bacteriological
Analytical Manual
Chapter 10, 2003
Method 1682
Method 1682
SM 9260 B
SM 9260 B
SM 9260 E
SM 9260 E
SM9213
TBD
SM 9260 H
SM 9260 H
SM 9260 I
SM 9260 I
FDA/Bacteriological
Analytical Manual
Chapter 10, 2003
FDA/Bacteriological
Analytical Manual
Chapter 10, 2003
Method 1682
Method 1682
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
TBD
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
TBD
Adapted from
analytical method
Adapted from
analytical method
SM 9260 I
SM 9260 I
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
SM 9260 B
SM 9260 B
SM 9260 E
SM 9260 E
SM9213
TBD
SM 9260 H
SM 9260 H
SM 9260 I
SM 9260 I
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
SM 9260 B
SM 9260 B
SM 9260 E
SM 9260 E
SM9213
TBD
SM 9260 H
SM 9260 H
Unlikely to be viable
Unlikely to be viable
Adapted from
analytical method
Adapted from
analytical method
Unlikely to be viable
Unlikely to be viable
Unlikely to be viable
Unlikely to be viable
Unlikely to be viable
Unlikely to be viable
Adapted from
analytical method
TBD
Unlikely to be viable
Unlikely to be viable
LRN
This pathogen is included in the HHS/USDA select agent list. Select agent methods are generally provided and controlled
by the LRN. In some cases, methods that apply to environmental samples may not be fully developed or validated.
If analysis of this agent is required, contact the LRN at (404) 639-2790 for information on the LRN laboratory capable of
receiving and processing the sample.
Viruses
Adeno viruses:
Enteric and non-enteric (A-F)
Yes
No
Tissue culture3
Real-time PCR
AEM 71(6): 3131-
3136
AEM 71(6): 3131-
3136
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
SAM Revision 3.1, Appendix C
C-2
November 15, 2007
-------
Pathogen (s)
Astroviruses
Caliciviruses: Noroviruses (NoV)
Caliciviruses: Sapovirus (SaV)
Coronaviruses:
SARS-associated human coronavirus
(SARS-HCoV)
Hepatitis E virus (HEV)
Influenza H5N1 virus
Orthopoxviruses: Variola, Vaccinia, Cowpox,
Monkeypox, Camelpox, Ectromelia, and
Gerbilpox
Picornaviruses: Enteroviruses
Picornaviruses:
Hepatitis A virus (HAV)
Reoviruses:
Rotavirus (Group A)
Viability
Yes
No
No
No
No
No
No
Yes
No
Yes
No
Yes
No
Yes
No
Analytical
Technique
Integrated Cell
Culture/RT-PCR
Real-time RT-PCR
Real-time RT-PCR
Real-time RT-PCR
Real-time RT-PCR
Real-time RT-PCR
Real-time RT-PCR
Tissue culture
PCR/lmmunoassay
Tissue culture
RT-PCR
Tissue culture
RT-PCR
Tissue culture
RT-PCR
Analytical
Method
Canadian Journal of
Microbiology 50:
269-278
Canadian Journal of
Microbiology 50:
269-278
Journal of Clinical
Microbiology 42(10):
4679-4685
Journal of Medical
Virology 78(10):
1347-1353
Journal of Virological
Methods 122: 29-36
Journal of Virological
Methods 131(1 ): 65-
71
Emerging Infectious
Diseases 11(8):
1303-1305
Solid1
(soil, powder)
Adapted from
analytical method4
Adapted from
analytical method4
Journal of Clinical
Microbiology Vol.
42(10): 4679-468S4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Particulate2
(swabs, wipes,
HEPA)
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Liquid/Water
(filter, grab)
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Drinking Water
(filter, grab)
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Aerosol
(growth media,
filter, liquid)
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
LRN
This pathogen is included in the HHS/USDA select agent list. Select agent methods are generally provided and controlled
by the LRN. In some cases, methods that apply to environmental samples may not be fully developed or validated.
If analysis of this agent is required, contact the LRN at (404) 639-2790 for information on the LRN laboratory capable of
receiving and processing the sample.
USEPA Manual of
Methods for Virology
EPA/600/4-84/013,
April 2001
AEM 69(6): 3158-
3164
TBD
AEM 69(6): 3158-
3164
AEM 69(6): 3158-
3164
AEM 69(6): 3158-
3164
Adapted from
analytical method4
Adapted from
analytical method4
TBD
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
TBD
Adapted from
analytical method4
Adapted from
analytical method4
Adapted from
analytical method4
USEPA Manual of
Methods for Virology
EPA/600/4-84/013,
April 2001
Adapted from
analytical method4
TBD
AEM Vol. 69 No. 6:
31 58-31 644
AEM Vol. 69 No. 6:
31 58-31 644
AEM Vol. 69 No. 6:
31 58-31 644
USEPA Manual of
Methods for Virology
EPA/600/4-84/013,
April 2001
Adapted from
analytical method4
TBD
AEM Vol. 69 No. 6:
31 58-31 644
AEM Vol. 69 No. 6:
31 58-31 644
AEM Vol. 69 No. 6:
31 58-31 644
Unlikely to be viable
Adapted from
analytical method4
TBD
Adapted from
analytical method4
Unlikely to be viable
Adapted from
analytical method4
Protozoa
Cryptosporidium spp.
[Cryptosporidiosis]
Yes
No
Tissue culture
I MS/FA and/or I FA
AEM 65(9): 3936-
3941
Method 1622 and/or
Method 1623
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Method 1623
Adapted from
analytical method
Method 1622
Unlikely to be found
Unlikely to be found
SAM Revision 3.1, Appendix C
C-3
November 15, 2007
-------
Pathogen (s)
Entamoeba histolytica
Giardiaspp.
Toxoplasma gondii [Toxoplasmosis]
Viability
Yes
No
Yes
No
Yes
No
Analytical
Technique
Culture
PCR
Culture
IMS/FA
Animal infectivity
PCR
Analytical
Method
Journal of
Parasitology 58(2):
306-310
Journal of Clinical
Microbiology
43(11): 5491 -5497
Trans. R. Soc. Trop.
Med. Hyg. 77(4):
487-488
Method 1623
Emerging Infectious
Diseases 12(2):
326-329
AEM 70(7): 4035-
4039
Solid1
(soil, powder)
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Particulate2
(swabs, wipes,
HEPA)
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Liquid/Water
(filter, grab)
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Method 1623
Emerging Infectious
Diseases Vol. 12
No. 2: 326-329
AEM Vol. 70 No. 7:
4035-4039
Drinking Water
(filter, grab)
Adapted from
analytical method
Adapted from
analytical method
Adapted from
analytical method
Method 1623
Emerging Infectious
Diseases Vol. 12
No. 2: 326-329
AEM Vol. 70 No. 7:
4035-4039
Aerosol
(growth media,
filter, liquid)
Unlikely to be found
Unlikely to be found
Unlikely to be found
Unlikely to be found
Unlikely to be found
Unlikely to be found
Helminths
Baylisascaris procyonis
Yes
No
Embryonation of
eggs
Microscopy
EPA/625/R92/013
EPA/625/R92/013
EPA/625/R92/013
EPA/625/R92/013
Adapted from
analytical method
Adapted from
analytical method
EPA/625/R92/013
EPA/625/R92/013
Adapted from
analytical method
Adapted from
analytical method
Unlikely to be found
Unlikely to be found
1 Solid samples (except those containing viruses) should be (1) prepared for culture according to EPA Method 1680, Fecal Conforms in Sewage Sludge (Biosolids) by Multiple-Tube Fermentation using Lauryl
Tryptose Broth (LTB) and EC Medium, and (2) prepared for PCR according to Quantification of Bias Related to the Extraction ofDNA Directly from Soils, Frostegard A, Courtois S, Ramisse V, Clerc S, Bernillon D,
Le Gall F, Jeannin P, Nesme X, Simonet P, AEM. 2005. 65(12): 5409-5420.
2 Particulate samples (except those containing viruses) should be prepared for culture according to Swab Materials and Bacillus anthracis Spore Recovery from Nonporous Surfaces, Rose, L, Jensen, B.,
Peterson, A., Banerjee, S.N., and M.J. Arduino, Emerg. Infec. Diseases. 2004 10(6): 1023-1029.
3 Given that adenovirus 40 and 41 can be difficult to grow in culture, additional cell lines such as G293 (Journal of Medical Virology. 1983. 11(3) :215-231) or CaCo-2 (Journal of Medical Virology. 1994. 44(3): 310-
315) may be considered when these viruses are suspected to be present.
4 Samples should be prepared according to procedures found in USEPA Manual of Methods for Virology EPA/600/4-84/013, April 2001.
SAM Revision 3.1, Appendix C
C-4
November 15, 2007
-------
Appendix D - SelectedBiotoxin Methods
Appendix D: Selected Biotoxin Methods
SAM Revision 3.1 November 15, 2007
-------
Appendix D - SelectedBiotoxin Methods
SAM Revision 3.1 November 15, 2007
-------
Appendix D: Biotoxin Methods
Analyte(s)
CASRN
Analysis Type
Analytical
Technique
Aerosol
(filter/cassette, liquid
impinger)
Solid
(soil, powder)
Particulate
(swabs, wipes, filters)
Liquid/Drinking Water
Protein
Abrin
Botulinum neurotoxins
(Serotoypes A, B, E, F)
a-Conotoxin
Ricin
Shiga and Shiga-like
Toxins
(Stx, Stx-1,Stx-2)
1393-62-0
NA
156467-85-5
9009-86-3
75757-64-1
Presumptive
Confirmatory
Biological Activity
Presumptive
Confirmatory
Biological Activity
Presumptive
Confirmatory
Biological Activity
Presumptive
Confirmatory
Biological Activity
Presumptive
Confirmatory
Biological Activity
Immunoassay
Competitive binding
assay
Enzyme activity
Immunoassay
Immunoassay
(ELISA)
Mouse Bioassay
Immunoassay
HPLC-MS
Receptor binding
Immunoassay
Immunoassay
(ELISA)
Enzyme activity
Optical
immunoassay
Immunoassay
(ELISA)
Competitive binding
assay
Adapted from 1 19th AOAC
Annual Meeting & Exposition,
2005, p. 613
Adapted from Pharmacology &
Toxicology 88(5): 255-260
Adapted from Analytical
Biochemistry 357(2): 200-207
Adapted from Lateral flow
immunassay kits
Adapted from FDA
Bacteriological Analytical
Manual, Chapter 17
Adapted from FDA
Bacteriological Analytical
Manual, Chapter 17
Adapted from Biochemical
Journal 328: 245-250
Adapted from Journal of
Medicinal Chemistry
47(5): 1234-1241
Adapted from Biochemical
Journal 328: 245-250
Adapted from Lateral flow
immunassay kits
Adapted from Journal of Food
Protection 68(6): 1294-1301
Adapted from Analytical
Biochemistry 357(2): 200-207
Adapted from Journal of Clinical
Microbiology 45(10): 3377-3380
Adapted from FDA
Bacteriological Analytical
Manual, Appendix 1
Adapted from Pharmacology &
Toxicology 88(5): 255-260
Adapted from 1 19th AOAC
Annual Meeting & Exposition,
2005, p. 613
Adapted from Pharmacology &
Toxicology 88(5): 255-260
Adapted from Analytical
Biochemistry 357(2): 200-207
Adapted from 1 19th AOAC
Annual Meeting & Exposition,
2005, p. 613
Adapted from Pharmacology &
Toxicology 88(5): 255-260
Adapted from Analytical
Biochemistry 357(2): 200-207
Adapted from 119th AOAC
Annual Meeting & Exposition,
2005, p. 613
Adapted from Pharmacology &
Toxicology 88(5): 255-260
Adapted from Analytical
Biochemistry 357(2): 200-207
LRN
If analysis for this agent is required in solid, particulate, or liquid samples, contact the LRN at
(404) 639-2790 for information of the closest LRN laboratory capable of receiving and processing
the sample. The terms presumptive and confirmatory as used for LRN methods are described in
Section 8.2.1.
Adapted from Biochemical
Journal 328: 245-250
Adapted from Journal of
Medicinal Chemistry
47(5): 1234-1241
Adapted from Biochemical
Journal 328: 245-250
Adapted from Biochemical
Journal 328: 245-250
Adapted from Journal of
Medicinal Chemistry
47(5): 1234-1241
Adapted from Biochemical
Journal 328: 245-250
Adapted from Biochemical
Journal 328: 245-250
Adapted from Journal of
Medicinal Chemistry
47(5): 1234-1241
Adapted from Biochemical
Journal 328: 245-250
LRN
If analysis for this agent is required in solid, particulate, or liquid samples, contact the LRN at
(404) 639-2790 for information of the closest LRN laboratory capable of receiving and processing
the sample. The terms presumptive and confirmatory as used for LRN methods are described in
Section 8.2.1.
Adapted from Journal of Food
Protection 68(6): 1294-1301
Adapted from Analytical
Biochemistry 357(2): 200-207
Adapted from Journal of Clinical
Microbiology 45(10): 3377-3380
Adapted from FDA
Bacteriological Analytical
Manual, Appendix 1
Adapted from Pharmacology &
Toxicology 88(5): 255-260
Adapted from Journal of Food
Protection 68(6): 1294-1301
Adapted from Analytical
Biochemistry 357(2): 200-207
Adapted from Journal of Clinical
Microbiology 45(10): 3377-3380
Adapted from FDA
Bacteriological Analytical
Manual, Appendix 1
Adapted from Pharmacology &
Toxicology 88(5): 255-260
Adapted from Journal of Food
Protection 68(6): 1294-1301
Adapted from Analytical
Biochemistry 357(2): 200-207
Adapted from Journal of Clinical
Microbiology 45(10): 3377-3380
Adapted from FDA
Bacteriological Analytical
Manual, Appendix 1
Adapted from Pharmacology &
Toxicology 88(5): 255-260
SAM Revision 3.1, Appendix D
D-1
November 15, 2007
-------
Analyte(s)
Staphylococcal
enterotoxins
(SEB)
Staphylococcal
enterotoxins
(SEA, SEC)
CASRN
39424-53-8
37337-57-8,
39424-54-9
Analysis Type
Presumptive
Confirmatory
Biological Activity
Presumptive
Confirmatory
Biological Activity
Analytical
Technique
Immunoassay
TBD
TBD
Immunoassay
TBD
TBD
Aerosol
(filter/cassette, liquid
impinger)
Adapted from 993.06 (AOAC)
TBD
TBD
Adapted from 993.06 (AOAC)
TBD
TBD
Solid
(soil, powder)
Particulate
(swabs, wipes, filters)
Liquid/Drinking Water
LRN
If analysis for this agent is required in solid, particulate, or liquid samples, contact the LRN at
(404) 639-2790 for information of the closest LRN laboratory capable of receiving and processing
the sample. The terms presumptive and confirmatory as used for LRN methods are described in
Section 8.2.1.
TBD
TBD
Adapted from 993.06 (AOAC)
TBD
TBD
TBD
TBD
Adapted from 993.06 (AOAC)
TBD
TBD
TBD
TBD
Adapted from 993.06 (AOAC)
TBD
TBD
Small Molecule
Aflatoxin
(Type B1)
a-Amanitin
Anatoxin-a
Brevetoxins
(B form)
Cylindrospermopsin
Diacetoxyscirpenol
(DAS)
1402-68-2
23109-05-9
64285-06-9
98225-48-0
143545-90-8
2270-40-8
Presumptive
Confirmatory
Presumptive
Confirmatory
Presumptive
Confirmatory
Presumptive
Confirmatory
Presumptive
Confirmatory
Presumptive
Confirmatory
Immunoassay
HPLC-FL
Immunoassay
HPLC
ampero metric
detection
TBD
HPLC-FL
(precolumn
derivatization)
Immunoassay
HPLC-MS
TBD
HPLC-MS
Immunoassay
HPLC-MS
Adapted from 991.31 (AOAC)
Adapted from 994.08 (AOAC)
Adapted from Journal Food
Protection 68(6): 1294-1301
Adapted from Journal of
Chromatography
563(2): 299-311
TBD
Adapted from Biomedical
Chromatography 10: 46-47
Adapted from Environmental
Health Perspectives
110(2): 179-185
Adapted from Toxicon
43(4): 455-465
TBD
Adapted from FEMS
Microbiology Letters
216(2): 159-164
Adapted from International
Journal of Food Microbiology
6(1): 9-1 7
Adapted from Rapid
Communications in Mass
Spectrometry 20(9): 1422-1428
Adapted from 991 .31 (AOAC)
Adapted from 994.08 (AOAC)
Adapted from Journal Food
Protection 68(6): 1294-1301
Adapted from Journal of
Chromatography
563(2): 299-311
TBD
Adapted from Biomedical
Chromatography 10: 46-47
Adapted from Environmental
Health Perspectives
110(2): 179-185
Adapted from Toxicon
43(4): 455-465
TBD
Adapted from FEMS
Microbiology Letters
216(2): 159-164
Adapted from International
Journal of Food Microbiology
6(1): 9-1 7
Adapted from Rapid
Communications in Mass
Spectrometry 20(9): 1422-1428
Adapted from 991 .31 (AOAC)
Adapted from 994.08 (AOAC)
Adapted from Journal Food
Protection 68(6): 1294-1301
Adapted from Journal of
Chromatography
563(2): 299-311
TBD
Adapted from Biomedical
Chromatography 10: 46-47
Adapted from Environmental
Health Perspectives
110(2): 179-185
Adapted from Toxicon
43(4): 455-465
TBD
Adapted from FEMS
Microbiology Letters
216(2): 159-164
Adapted from International
Journal of Food Microbiology
6(1): 9-1 7
Adapted from Rapid
Communications in Mass
Spectrometry 20(9): 1422-1428
Adapted from 991 .31 (AOAC)
Adapted from 994.08 (AOAC)
Adapted from Journal Food
Protection 68(6): 1294-1301
Adapted from Journal of
Chromatography
563(2): 299-31 1
TBD
Adapted from Biomedical
Chromatography 10: 46-47
Adapted from Environmental
Health Perspectives
110(2): 179-185
Adapted from Toxicon
43(4): 455-465
TBD
Adapted from FEMS
Microbiology Letters
216(2): 159-164.
Adapted from International
Journal of Food Microbiology
6(1): 9-1 7
Adapted from Rapid
Communications in Mass
Spectrometry 20(9): 1422-1428
SAM Revision 3.1, Appendix D
D-2
November 15, 2007
-------
Analyte(s)
Microcystins
(Principal isoforms: LR,
YR, RR, LW)
Picrotoxin
Saxitoxin
(SIX, NEOSAX, GTX,
dcGTX, dcSTX)
T-2 Mycotoxin
Tetrodotoxin
CASRN
77238-39-2
124-87-8
35523-89-8
21259-20-1
9014-39-5
Analysis Type
Presumptive
Confirmatory
Presumptive
Confirmatory
Presumptive
Confirmatory
Presumptive
Confirmatory
Presumptive
Confirmatory
Analytical
Technique
Immunoassay/
Phosphatase assay
HPLC-MS
Immunoassay
HPLC
Immunoassay
HPLC-FL
(post column
derivatization)
Immunoassay
HPLC-MS
Immunoassay
HPLC-MS
Aerosol
(filter/cassette, liquid
impinger)
Adapted from Journal of AOAC
International 84(4): 1035-1044
Adapted from Analyst
119(7): 1525-1530
TBD
Adapted from Journal of
Pharmaceutical and Biomedical
Analysis 7(3): 369-375
TBD
Adapted from Journal of AOAC
International 78(2): 528-532
Adapted from Journal Food
Protection 68(6): 1294-1301
Adapted from Rapid
Communications in Mass
Spectrometry 20(9): 1422-1428
Adapted from Journal of Clinical
Laboratory Analysis 6(2): 65-72
Adapted from Analytical
Biochemistry 290(1): 10-17
Solid
(soil, powder)
Adapted from Journal of AOAC
International 84(4): 1035-1044
Adapted from Analyst
119(7): 1525-1530
TBD
Adapted from Journal of
Pharmaceutical and Biomedical
Analysis 7(3): 369-375
TBD
Adapted from Journal of AOAC
International 78(2): 528-532
Adapted from Journal Food
Protection 68(6): 1294-1301
Adapted from Rapid
Communications in Mass
Spectrometry 20(9): 1422-1428
Adapted from Journal of Clinical
Laboratory Analysis 6(2): 65-72
Adapted from Analytical
Biochemistry 290(1): 10-17
Particulate
(swabs, wipes, filters)
Adapted from Journal of AOAC
International 84(4): 1035-1044
Adapted from Analyst
119(7): 1525-1530
TBD
Adapted from Journal of
Pharmaceutical and Biomedical
Analysis 7(3): 369-375
TBD
Adapted from Journal of AOAC
International 78(2): 528-532
Adapted from Journal Food
Protection 68(6): 1294-1301
Adapted from Rapid
Communications in Mass
Spectrometry 20(9): 1422-1428
Adapted from Journal of Clinical
Laboratory Analysis 6(2): 65-72
Adapted from Analytical
Biochemistry 290(1): 10-17
Liquid/Drinking Water
Adapted from Journal of AOAC
International 84(4): 1035-1044
Adapted from Analyst
119(7): 1525-1530
TBD
Adapted from Journal of
Pharmaceutical and Biomedical
Analysis 7(3): 369-375
TBD
Adapted from Journal of AOAC
International 78(2): 528-532
Adapted from Journal Food
Protection 68(6): 1294-1301
Adapted from Rapid
Communications in Mass
Spectrometry 20(9): 1422-1428
Adapted from Journal of Clinical
Laboratory Analysis 6(2): 65-72
Adapted from Analytical
Biochemistry 290(1): 10-17
SAM Revision 3.1, Appendix D
D-3
November 15, 2007
-------
Revision 3.1, Appendix D D - 4 November 15, 2007
-------
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sited
ivironmental Protection
jency
fiecfcfcd/iecfclable
>-" tinted ink on
paper that contains a minimum of
50%
psv/cessed chlorine
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