&EPA
United States
Environmental Protection
Agency
Standardized Analytical Methods for
Use During Homeland Security Events
Revision 1.0
September 29, 2004
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EPA/600/R-04/126
September 2004
for
1.0
Prepared by
DynCorp Systems & Solutions LLC (A CSC Company)
Chantilly, VA20151
EPA Contract No. 68-W-01-034
Work Assignment Manager
Oba Vincent
National Homeland Security Research Center
Office of Research and Development
Cincinnati, OH 45268
National Homeland Security Research Center
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, OH 45268
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Disclaimer
The U.S. Environmental Protection Agency through its Office of Research and Development funded and
managed the research described here under Contract 68-W-01-034 to DynCorp Systems & Solutions LLC
(A CSC Company). It has been subjected to the Agency's peer and administrative review and has been
approved for publication as an EPA document. Mention of trade names or commercial products 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)
26 West Martin Luther King Drive
Cincinnati, OH 45268
(513)569-7456
vincent. oba@epa. gov
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Foreword
The U.S. Environmental Protection Agency is charged by Congress with protecting the Nation's land, air,
and water resources. Under a mandate of national environmental laws, the Agency 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 science knowledge 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 the Agency's center for conducting
research to facilitate protection and decontamination of structures and water infrastructure subject to
chemical or biological terror attacks. NHSRC's research is designed to provide appropriate, affordable,
effective, and validated technologies, methods, and guidance to understand the risk posed by potential
chemical and biological terror attacks on buildings and water infrastructure, and to enhance our ability to
detect, contain, and clean up in the event of such attacks. NHSRC will also provide direct technical
assistance to response personnel in the event of future deliberate physical and radiological,
chemical/biological attacks on buildings or water infrastructure and provide interagency liaison for EPA
in Homeland Security research and technology.
This publication has been produced as part of the Center'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.
E. Timothy Oppelt, Director
National Homeland Security Research Center
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Acknowledgments
The contributions of the following persons and organizations to the development of this document are
gratefully acknowledged:
EPA Homeland Security Laboratory Capability Workgroup Analytical Methods Subteam
Steve Allgeier, Office of Water
David Friedman, Office of Research and Development
Ben Hull, Office of Indoor Air and Radiation
Michael S. Johnson, Office of Solid Waste and Emergency Response
Alan Lindquist, Office of Research and Development
Oba Vincent, Office of Research and Development
Environmental Protection Agency
Linda Anderson-Carnahan, Region 10 - Manchester Laboratory
Don Betowski, National Exposure Research Laboratory
Isa Chamberlain, Region 10 - Manchester Laboratory
Joe Dorsey, Region 3 - Environmental Sciences Center
Diane Gregg, Region 6 Laboratory - Houston Branch
Ann C. Grimm, Office of Research and Development
Stephanie Harris, Region 10 - Manchester Laboratory
Jafirul Hasan, Health and Ecological Criteria Division
Bonita Johnson, Region 4 - Science and Ecosystems Support Division
Tammy Jones-Lepp, National Exposure Research Laboratory
Irwin Katz, Region 2
Peggy Knight, Region 10 - Manchester Laboratory
Barry Lesnik, Office of Solid Waste
John Nebelsick, Office of Solid Waste and Emergency Response
Ed O'Neill, Region 6 Laboratory - Houston Branch
Steve Reimer, Region 10 - Manchester Laboratory
Lavon Revells, Region 4 - Science and Ecosystems Support Division
Eugene Rice, Office of Research and Development
Fred Siegelman, Office of Pesticides Programs - Biological and Economic Analysis Division
Terry Smith, Office of Solid Waste and Emergency Response
Dave Stockton, Region 6 Laboratory - Houston Branch
Sue Warner, Region 3 - Environmental Sciences Center
Laura Webb, Region 7 - ENSVRLAB
Wayne Whipple, Region 5
Sen-yi Yang, Office of Solid Waste and Emergency Response
United States Geological Survey
Merle Shockey
IV
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Centers for Disease Control and Prevention
Matthew J. Arduino, Division of Health Quality Promotion
Jay E. Gee, Division of Bacterial and Mycotic Diseases
Dennis Juranek, Division of Parasitic Diseases
Laura J. Rose, Division of Health Quality Promotion
Department of Homeland Security
Lance Brooks
Federal Bureau of Investigation
Ben Garrett, Hazard Materials Response Unit
Paul Keller, Hazard Materials Response Unit
Jarrad Wagner, Hazard Materials Response Unit
Food and Drug Administration
David L. Craft, Molecular Biology and Microbiology
United States Army Edgewood Chemical Biological Center
Dennis Reutter, Edgewood Chemical Biological Center
John D. Wright, Dugway Proving Ground, Life Sciences Division
United States Department of Agriculture
Ronald Payer
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Standardized Analytical Methods for Use
During Homeland Security Events
Revision 1.0
September 2004
Contents
Disclaimer ii
Foreword iii
Acknowledgments iv
Section 1.0: Introduction 1
Section 2.0: Scope and Application 5
Section 3.0: Points of Contact 7
Section 4.0: Chemical Methods 9
4.1 General Guidance 9
4.1.1 Standard Operating Procedures for Identifying Chemical Methods 9
4.1.2 General Quality Control (QC) Guidance for Chemical Methods 10
4.1.3 Safety and Waste Management 11
4.2 Method Summaries 12
4.2.1 Laboratory Response Network (LRN) 12
4.2.2 CLP Method SOW ILM05.3 Cyanide: Analytical Methods for Total
Cyanide Analysis 13
4.2.3 NERL Method 365.1, Revision 2: Determination of Phosphorus by Semi-
Automated Colorimetry 13
4.2.4 EPA Method 200.2, Revision 2.8: Sample Preparation Procedure for
Spectrochemical Determination of Total Recoverable Elements 14
4.2.5 EPA Method 300.1: Determination of Inorganic Anions in Drinking Water
by Ion Chromatography 14
4.2.6 EPA Method 549.2: Determination of Diquat and Paraquat in Drinking
Water by Liquid-Solid Extraction and High-Performance Liquid Chromatography
with Ultraviolet Detection 15
4.2.7 EPA Method 3031 (SW-846): Acid Digestion of Oils for Metals Analysis by
Atomic Absorption or ICP Spectrometry 15
4.2.8 EPA Method 3050B (SW-846): Acid Digestion of Sediments, Sludges,
and Soils 15
4.2.9 EPA Method 3520C (SW-846): Continuous Liquid-Liquid Extraction 16
4.2.10 EPA Method 3535A (SW-846): Solid-Phase Extraction 18
4.2.11 EPA Method 3541 (SW-846): Automated Soxhlet Extraction 19
4.2.12 EPA Method 3545A (SW-846): Pressurized Fluid Extraction (PFE) 21
4.2.13 EPA Method 3580A (SW-846): Waste Dilution 23
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4.2.14 EPA Method 3585 (SW-846): Waste Dilution for Volatile Organics 24
4.2.15 EPA Method 5030C (SW-846): Purge-and-Trap for Aqueous Samples 25
4.2.16 EPA Method 5035A (SW-846): Closed-System Purge-and-Trap and
Extraction for Volatile Organics in Soil and Waste Samples 25
4.2.17 EPA Method 5050 (SW-846): Bomb Preparation Method for Solid Waste 26
4.2.18 EPA Method 6010C (SW-846): Inductively Coupled Plasma - Atomic
Emission Spectrometry 26
4.2.19 EPA Method 6020A (SW-846): Inductively Coupled Plasma - Mass
Spectrometry 27
4.2.20 EPA Method 7010 (SW-846): Graphite Furnace Atomic Absorption
Spectrophotometry 27
4.2.21 EPA Method 7470A (SW-846): Mercury in Liquid Wastes (Manual
Cold-Vapor Technique) 28
4.2.22 EPA Method 747IB (SW-846): Mercury in Solid or Semisolid Wastes
(Manual Cold-Vapor Technique) 28
4.2.23 EPA Method 8015C (SW-846): Nonhalogenated Organics Using GC/FID .... 28
4.2.24 EPA Method 8260B (SW-846): Volatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS) 29
4.2.25 EPA Method 8270D (SW-846): Semivolatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS) 30
4.2.26 EPA Method 8315A (SW-846): Determination of Carbonyl Compounds
by High Performance Liquid Chromatography (HPLC) 31
4.2.27 EPA Method 83ISA (SW-846): 7V-Methylcarbamates by High Performance
Liquid Chromatography (HPLC) 32
4.2.28 EPA Method 832IB (SW-846): Solvent-Extractable Nonvolatile
Compounds by High Performance Liquid Chromatography/Thermospray/
Mass Spectrometry (HPLC/TS/MS) or Ultraviolet (UV) Detection 32
4.2.29 AOAC Official Method 994.08: Aflatoxin in Corn, Almonds, Brazil Nuts,
Peanuts, and Pistachio Nuts 33
4.2.30 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 33
4.2.31 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 34
4.2.32 ISO Method - 10312: Ambient Air - Determination of Asbestos Fibres -
Direct-transfer Transmission Electron Microscopy Method (TEM) 34
4.2.33 NIOSH Method 6001: Arsine 35
4.2.34 NIOSH Method 6002: Phosphine 35
4.2.35 NIOSH Method 6004: Sulfur Dioxide 35
4.2.36 NIOSH Method 6010: Hydrogen Cyanide 36
4.2.37 NIOSH Method 6011: Bromine and Chlorine 36
4.2.38 NIOSH Method 6013: Hydrogen Sulfide 36
4.2.39 NIOSH Method 6015: Ammonia 37
4.2.40 NIOSH Method 6402: Phosphorus Trichloride 37
4.2.41 NIOSH Method 7903: Acids, Inorganic 37
4.2.42 NIOSH Method 7904: Cyanides, Aerosol and Gas 38
4.2.43 NIOSH Method 7906: Fluorides, Aerosol and Gas 38
4.2.44 NIOSH Method S301-1: Fluoroacetate Anion 39
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4.2.45 OSHA Method ID-188: Ammonia in Workplace Atmospheres-Solid Sorbent . 39
4.2.46 OSHA Method ID-216SG: Boron Trifluoride (BF3) 39
4.2.47 Standard Method 4110 B: Ion Chromatography with Chemical Suppression
of Eluent Conductivity 40
4.2.48 Standard Method 4500-NH3 B: Preliminary Distillation Step 40
4.2.49 Standard Method 4500-NH3 G: Automated Phenate Method 40
4.2.50 Standard Method 4500-C1 G: DPD Colorimetric Method 41
4.2.51 IO Compendium Method IO-3.1: Selection, Preparation, and Extraction of
Filter Material 41
4.2.52 IO Compendium Method IO-3.4: Determination of Metals in Ambient
Particulate Matter Using Inductively Coupled Plasma (ICP) Spectroscopy .... 42
4.2.53 IO Compendium Method IO-3.5: Determination of Metals in Ambient
Particulate Matter Using Inductively Coupled Plasma/Mass Spectrometry
(ICP/MS) 42
4.2.54 IO Compendium Method IO-5: Sampling and Analysis for Vapor and
Particle Phase Mercury in Ambient Air Utilizing Cold Vapor Atomic
Fluorescence Spectrometry (CVAFS) 43
4.2.55 EPA Air Toxics Method - 6 (TO-6): Method for the Determination of
Phosgene in Ambient Air Using High Performance Liquid Chromatography ... 43
4.2.56 EPA Air Toxics Method - 13A (TO-13A): Determination of Fob/cyclic
Aromatic Hydrocarbons (PAHs) in Ambient Air Using Gas Chromatography/
Mass Spectrometry (GC/MS) 44
4.2.57 EPA Air Toxics Method - 15 (TO-15): Determination of Volatile
Organic Compounds (VOCs) in Air Collected in Specially-Prepared
(Summa) Canisters and Analyzed by Gas Chromatography/
Mass Spectrometry (GC/MS) 45
4.2.58 Journal of Analytical Atomic Spectrometry, 2000, 15, pp. 277-279: Boron
Trichloride Analysis 46
4.2.59 Analytical Letters, 1994, 27 (14), pp. 2703-2718: Screening-Procedure for
Sodium Fluoracetate (Compound 1080) at Sub-Microgram/Gram
Concentrations in Soils 46
Section 5.0: Biological Methods 47
5.1 General Guidance 47
5.1.1 Standard Operating Procedures for Identifying Biological Methods 48
5.1.2 General Quality Control (QC) Guidance for Biological Methods 49
5.1.3 Safety and Waste Management 49
5.2 Method Summaries 50
5.2.1 Laboratory Response Network (LRN) 50
5.2.2 Biosafety Level 4 Viruses 51
5.2.3 U.S. EPA Method 1622: Cryptosporidium in Water by Filtration/IMS/FA 51
5.2.4 ICRMicrobial Laboratory Manual Chapter XI 52
5.2.5 USDA/FSIS 4 52
5.2.6 Standard Methods 9260 E: Shigella spp 53
5.2.7 Standard Methods 9260 F: Pathogenic Escherichia coli 53
5.2.8 Standard Methods 9260 G: Campylobacter jejuni 54
5.2.9 Standard Methods 9260 H: Vibrio cholerae 54
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5.2.10 Literature Reference for Caliciviruses 55
5.2.11 Literature Reference for Enteroviruses 55
5.2.12 Literature Reference for Hepatitis A Viruses 56
5.2.13 Literature Reference for Venezeulan Equine Encephalitis (VEE) Virus 57
5.2.14 Literature Reference for Toxoplasma gondii 57
5.2.15 Entamoeba histolytica. Filtration/IMS/PCR 58
Section 6.0: Conclusions 59
Abbreviations and Acronyms 61
Appendices
A Chemical Methods A-l
B Biological Methods B-l
Figures
1-1. Analytical Response Roadmap for Homeland Security Events 2
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Standardized Analytical Methods for Use During Homeland Security Events
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 successfully 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 incidents, the Environmental Protection Agency's
(EPA) 9/77 Lessons Learned Report and its Anthrax Lessons Learned report identified several areas
where the country could better prepare itself in the event of future incidents. One of the most important
areas was the need to improve the nation's laboratory capacity and capability to respond to incidents
requiring the analysis of large numbers of environmental samples in a short time.
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, cross-cutting laboratory issues. The EPA Homeland Security Laboratory Capacity
Workgroup consists of representatives from the Office of Research and Development, Office of Radiation
and Indoor Air, Office of Water, Office of Solid Waste and Emergency Response, Office of
Environmental Information, Office of Pesticide Programs, EPA Region 1, EPA Region 2, EPA Region 4,
and EPA Region 6.
A critical area identified by the workgroup was the need for a list of pre-selected, pre-evaluated,
standardized analytical methods to be used by all laboratories when analyzing homeland security incident
samples. 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, analyzing data
and making decisions.
To this end, workgroup members formed an Analytical Methods Subteam to address homeland security
methods issues. Figure 1-1 represents the analytical decision tree for responding to homeland security
incidents.
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Standardized Analytical Methods for Use During Homeland Security Events
Determination
Type of samples include
Risk assessment
Remediation
Clearance
~l
The "Determination" phase of environmental sample analysis in
response to a homeland security event is discussed within this
document. Other phases will be addressed in a separate document.
Figure 1-1. Analytical Response Roadmap for Homeland Security Events
The Subteam focused first on methods for use in assessing the extent of contamination and the efficacy of
decontamination. A survey of available analytical methods for approximately 120 biological and chemical
analytes was conducted using existing method resources including the following:
• National Environmental Methods Index (NEMI)
• Environmental Monitoring Method Index (EMMI)
• EPA Test Methods Index
EPA Office of Solid Waste SW-846 Methods On-line
EPA Microbiology Methods
National Institute for Occupational Safety and Health (NIOSH) method index
Occupational Safety and Health Administration (OSHA) method index
AOAC International
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Standardized Analytical Methods for Use During Homeland Security Events
• ASTM International
• International Organization for Standardization (ISO) methods
• Standard Methods for the Examination of Water and Wastewater
PubMED Literature Database
EPA's National Homeland Security Research Center brought together experts from across EPA and from
its sister agencies in a workshop setting to develop a compendium of analytical methods to be used when
responding to future incidents. The methodology was considered for both chemical and biological agents
of concern in the types of environmental sample matrices that were anticipated for analysis in homeland
security incidents. The primary objective of this effort was not to identify the "best" method, but rather to
have a balanced approach between leveraging existing determinative techniques and methodologies and
providing consistent analytical results.
These EPA Homeland Security Biological and Chemical Method Review and Consolidation Workshops
were held on April 13, 2004, and April 14, 2004, respectively. Participants included representatives from
the EPA program offices, the EPA regions, the Centers for Disease Control and Prevention (CDC), the
Food and Drug Administration (FDA), the Department of Homeland Security (DHS), the Federal Bureau
of Investigation (FBI), the Department of Defense (DOD), the U.S. Department of Agriculture (USDA),
and the U.S. Geological Survey (USGS).
During the workshops, participants were provided worksheets populated with as many known techniques
as possible to select a single determinative technique and analytical method for each analyte. For
biological methods, sample preparation techniques and/or sampling procedures were identified for water,
dust, and air matrices. Workshop participants identified sample preparation procedures for chemical
contaminants in solid, oily solid, aqueous, liquid, and gas phase matrices.
Not all methods identified and selected by workshop participants have been validated for the
analyte/matrix combinations of concern. However, this is a living document, and the Agency anticipates
that it will be updated periodically to reflect advances in analytical methodology, development of new
technologies, and changes in analytes based on needs. The Agency also anticipates that standardized
analytical protocols will be developed as needed, and that addendums may be generated to address
materials that are not included in this document.
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Standardized Analytical Methods for Use During Homeland Security Events
Section 2.0: Scope and Application
The purpose of this document is to identify and briefly describe the specific methodologies that EPA and
its contractors will employ when called upon to analyze environmental samples in times of national
emergency. The methodologies presented in this document should be used to:
• Evaluate the nature and extent of contamination
Evaluate decontamination effectiveness
The list of methods is limited to those methods that would be used to determine, to the extent possible
within analytical limitations, the presence of chemical and biological agents of concern and to determine
their concentrations in environmental media. The methods are not designed to be used for conducting an
initial evaluation (triage) of suspected material to determine if it poses an immediate danger or if it needs
to be analyzed in specially designed, highly secure facilities. Methods for addressing this analytical need
are and will be the subject of other efforts. It is hoped that this document will also assist State and local
governments in preparing for future emergencies.
Any deviations from the methods referenced in this document should be coordinated with the appropriate
point(s) of contact identified in Section 3.
Participants in the EPA Homeland Security Biological and Chemical Method Review and Consolidation
Workshops evaluated the suitability of existing methodologies and selected this set of methods for use by
the EPA laboratories and contract laboratories if called upon to respond to an emergency. The Agency
recognizes, however, that this advanced selection of such methods poses potential risks. These include the
following:
• Selecting technologies that may not be the most cost-effective technologies that are 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 the Agency did not anticipate having to analyze for a particular analyte or
analyte/matrix combination
• Preventing development and adoption of new and better measurement technologies
To address these potential risks as soon as possible, the Agency 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 are geared to the data
quality needs of the particular stage of the emergency response/recovery process) for all
analyte/matrix combinations listed in this document
• Specifying minimum measurement system verification (e.g., ASTM Standard D6956-03) and
documentation standards for homeland security analyses
Working with other government agencies and the private sector to establish a laboratory
accreditation system to ensure that laboratories selected to assist the Agency and its Federal, State,
and local partners in responding to homeland security incidents have the requisite expertise and
systems to perform this type of testing
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Section 3.0: Points of Contact
Questions concerning this document should be addressed to the appropriate point(s) of contact identified
below. 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.
Prior to use of one of the methods listed in Appendix A (chemical) or Appendix B (biological) in
response to a homeland security event, the appropriate point(s) of contact identified below should be
consulted. Additionally, any deviations from the protocols should be coordinated with the appropriate
point(s) of contact listed below.
General
Oba Vincent
National Homeland Security Research Center
Office of Research and Development (163)
26 West Martin Luther King Drive
Cincinnati, OH 45268
(513)569-7456
vincent.oba@epa.gov
Chemical Methods
Michael S. Johnson
Office of Solid Waste and Emergency Response (5204G)
USEPA Headquarters Ariel Rios Building
1200 Pennsylvania Avenue, NW
Washington, DC 20460
(703) 603-0266
johnson.michaels(g),epa.gov
David Friedman
Office of Research and Development (8101R)
USEPA Headquarters Ariel Rios Building
1200 Pennsylvania Avenue, NW
Washington, DC 20460
(202) 564-6662
friedman.david@epa.gov
Biological Methods
Alan Lindquist
National Homeland Security Research Center
Office of Research and Development (163)
26 West Martin Luther King Drive
Cincinnati, OH 45268
(513)569-7192
lindauist.alan@,epa.gov
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Standardized Analytical Methods for Use During Homeland Security Events
Eugene Rice
National Homeland Security Research Center
Office of Research and Development (163)
26 West Martin Luther King Drive
Cincinnati, OH 45268
(513)569-7204
rice.eugene@epa.gov
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Standardized Analytical Methods for Use During Homeland Security Events
Section 4.0: Chemical Methods
A list of analytical methods to be used in analyzing environmental samples for chemical contaminants
during homeland security events is provided in Appendix A. Methods are listed for each analyte and for
each sample matrix that potentially may need to be measured and analyzed when responding to an
emergency. The methods table in Appendix A is sorted alphabetically by analyte and includes the
following information:
Analyte(s). The compound or compound(s) of interest.
• CAS Number. 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.
• Determinative method identifier. The unique identifier or number assigned to an analytical method
by the method publisher.
Solid phase sample preparation procedure. The recommended method/procedure for sample
preparation to measure the analyte of interest in solid phase samples.
Oily solid sample preparation procedure. The recommended method/procedure for sample
preparation to measure the analyte of interest in oily phase samples.
• Aqueous/Liquid phase sample preparation procedure. The recommended method/procedure for
sample preparation to measure the analyte of interest in aqueous and/or liquid phase samples.
• Gas phase sample preparation procedure. The recommended method/procedure for sample
preparation and analysis to measure the analyte of interest in gas phase samples.
4.1 General Guidance
The guidance summarized in this section provides a general overview of how to identify the appropriate
chemical method(s) for a given analyte-matrix combination as well as recommendations for quality
control procedures.
For additional information on the properties of the chemicals listed in Appendix A, TOXNET
(http://toxnet.nlm.nih.gov/index.html'). a cluster of databases on toxicology, hazardous chemicals, and
related areas maintained by the National Library of Medicine, is an excellent resource. Additional
research on chemical contaminants is ongoing within EPA, and databases to manage this information are
currently under development.
4.1.1 Standard Operating Procedures for Identifying Chemical Methods
To determine the appropriate method and sample preparation technique that is to be used on the
environmental samples, locate analyte of concern in Appendix A: Chemical Methods under the "Analyte"
column. After locating the analyte of concern, continue across the table to identify the determinative
technique and determinative method for that particular compound. To determine the sample preparation
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Standardized Analytical Methods for Use During Homeland Security Events
technique select the appropriate matrix column (Solid Phase, Oily Solid, Aqueous/Liquid Phase, or Gas
Phase) for that particular analyte.
Sections 4.2.1 through 4.2.59 below provide summaries of the determinative and sample preparation
methods listed in Appendix A. Where available, a direct link to the full text of the selected analytical
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 1.
Table 1. 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, 20th Edition
Annual Book of ASTM Standards
International Organization for
Standardization Methods
Official Methods of Analysis of AOAC
International
Publisher
EPA, USGS
EPA Office of Water
EPA Office of Solid Waste
EPA Office of Research and
Development
EPA Office of Air and
Radiation
OSHA
NIOSH
American Public Health
Association and American
Water Works Association
ASTM International
ISO
AOAC International
Reference
http://vwwv.nemi.qov
http://www.epa.qov/safewater/methods/
sourcalt.html
http://www.epa.qov/epaoswer/hazwaste
/test/main, htm
http://www.epa.gov/nerlcwww/ordmeth.
htm
http://www.epa.qov/ttn/amtic/airtox.html
http://www.osha-slc.qov/dts/sltc/method
s/toc.html
http://www.cdc.gov/niosh/nmam/
http://www.apha.org
http://www.awwa.org
ISBN: 0875532357
http://www.astm.org
http://www.iso.org
http://www.aoac.org
4.1.2 General Quality Control (QC) Guidance for Chemical Methods
Having data of known and documented quality is critical in order for public officials to accurately assess
how to respond to emergency situations. 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.
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While method-specific QC requirements are described in many of the individual methods that are cited in
this manual, and will be included in any standardized analytical protocols developed to address specific
analytes and matrices of concern, the following describes a minimum set of QC procedures that shall be
conducted for all chemical testing. Individual methods, sampling and analysis protocols, or contractual
statements of work also should be consulted to determine any additional QC that may be needed. These
QC requirements generally consist of analysis of laboratory control samples and or matrix spikes to
identify and quantify measurement system accuracy at the levels of concern, blanks as a measure of
freedom from contamination, and matrix spike duplicates (MSB) or sample replicates to assess data
precision. 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 analysis of standards and other samples to ensure the continued reliability of the analytical
results. Examples of sufficient quality control includes:
Demonstration that 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/matrix/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 3 should be consulted regarding
appropriate quality assurance and quality control procedures prior to sample analysis.
4.1.3 Safety and Waste Management
It is imperative that safety precautions are used during collection, processing, and analysis of
environmental samples, particularly in emergency response situations that may include unknown hazards.
Many of the methods summarized or cited in Section 4.2 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:
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)
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4.2 Method Summaries
Method summaries for the analytical methods listed in Appendix A, including methods for sample
preparation and determinative techniques, are provided in Sections 4.2.1 - 4.2.59. Information provided in
these sections contains summary information only, extracted from the selected methods. The full version
of the method should be consulted prior to sample analysis.
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 the full version of the method or source
for obtaining a full version of the method.
Please note that not all methods have been validated for the analyte/matrix combination listed in
Appendix A. Please refer to the specified method to identify analyte/matrix combinations that have been
validated. Any questions regarding information discussed in this section should be addressed to the
appropriate contact(s) listed in Section 3.
4.2.1 Laboratory Response Network (LRN)
The agents listed below should be analyzed in accordance with the appropriate LRN protocols:
Contaminants
Alpha amanitin
Botulinum toxin
Microcystin
Ricin
Tetanus Toxin
CAS Number
NA
NA
NA
9009-86-3
NA
These agents will be analyzed using restricted procedures available only through the Laboratory Response
Network (LRN). These procedures are not available to the general laboratory community and thus are not
discussed within this document. For additional information on the LRN, please see contact information
listed below or visit http://www.bt.cdc.gov/lrn/.
Centers for Disease Control and Prevention
Laboratory Response Branch
Bioterrorism Preparedness and Response Program
National Center for Infectious Diseases
1600 Clifton Road NE, Mailstop C-18
Atlanta, GA 30333
Telephone: (404) 639-2790
E-mail: lrn(gicdc.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).
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Association of Public Health Laboratories
2025 M Street NW, Suite 550
Washington, DC 20036
Telephone: (202) 822-5227
Fax: (202) 887-5098
Website: www.aphl.org
E-mail: mfo@aphl.org
4.2.2 CLP Method SOW ILM05.3 Cyanide: Analytical Methods for Total Cyanide Analysis
This method should be used for preparation of solid, oily-solid, and aqueous/liquid phase samples for the
contaminant identified below and listed in Appendix A:
Contaminant
Cyanide
CAS Number
57-12-5
The method allows for either large volume (500 mL aqueous/liquid samples or 1 - 5 g solid samples
mixed with 500 mL of reagent water) or medium volume (50 mL aqueous/liquid samples, or 1 g solid
samples mized with 50 mL 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 also interfere with the distillation procedure.
Source: http://www.epa.gov/superfund/programs/cb/download/ilm/ilm53d.pdf
4.2.3 NERL Method 365.1, Revision 2: Determination of Phosphorus by Semi-Automated
Colorimetry
This method should be used for preparation and analysis of aqueous/liquid phase samples for the
contaminant identified below and listed in Appendix A:
Contaminant
Red Phosphorus
CAS Number
7723-14-0
This method measures all forms of phosphorus present in the sample, converting them to orthophosphate.
The analyte is determined as a reduced antimony-phospho-molybdate complex. A 50-mL sample is
digested with sulfuric acid and ammonium persulfate. The digestate is analyzed by automated
spectrophotometry (colorimetry) in which the sample reacts with color agents. The range of the method is
0.01 - 1.0 mg P/L. Silica, arsenate, nitrite, and sulfide may cause interference.
Source: http://infotrek.er.usgs.gov/intermedia/nemi port read/mediaget/nemi get blob/23
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4.2.4 EPA Method 200.2, Revision 2.8: Sample Preparation Procedure for
Spectrochemical Determination of Total Recoverable Elements
This method should be used for preparation of aqueous/liquid phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Arsenic (III) compounds
Arsenic trichloride
Arsine
Cadmium
Metals, NOS
CAS Number
22569-72-8
7784-34-1
7784-42-1
7440-43-9
NA
For aqueous/liquid samples, a 100-mL aliquot of the sample is digested with nitric and hydrochloric
acids, the volume reduced to approximately 20 mL, and worked up to a final volume of 50 mL. Samples
to be analyzed by Method 6020A (SW-846) require additional dilution to reduce chloride interference.
Source: "Methods for the Determination of Metals in Environmental Samples," Supplement I, National
Exposure Risk Laboratory-Cincinnati (NERL-CI), EPA/600/R-94/11, May 1994; and "Methods for the
Determination of Inorganic Substances in Environmental Samples," NERL-CI, EPA/600/R-93/100,
August, 1993 are available from National Technical Information Service (NTIS), 5285 Port Royal Road,
Springfield, VA 22161. Phone: 800-553-6847.
4.2.5 EPA Method 300.1: Determination of Inorganic Anions in Drinking Water by Ion
Chromatography
This method should be used for the preparation and analysis of aqueous/liquid phase samples for the
contaminants identified below and listed in Appendix A:
Contaminant
Fluoracetate Salts
CAS Number
NA
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 comprised of 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
utilize 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 2 orders of magnitude.
Source: http://infotrek.er.usgs.gov/intermedia/nemi port read/mediaget/nemi get blob/159
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4.2.6 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 phase samples for the
contaminants identified below and listed in Appendix A:
Contaminant
Paraquat
CAS Number
4685-14-7
A 250-mL sample is extracted using a C-8 liquid/solid extraction (LSE) cartridge or a C-8 disk which 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 high performance liquid
chromatography (HPLC) system equipped with a UV absorbance detector. A photodiode array detector is
utilized to provide simultaneous detection and confirmation of the method analytes. The analytical range
depends on the sample matrix and the instrumentation used.
Source: http://www.epa.gov/nerlcwww/rn 549 2.pdf
4.2.7 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 oily-solid phase samples for the contaminants identified
below and listed in Appendix A:
Contaminant
Arsenic (III) compounds
Arsenic trichloride
Cadmium
Metals, NOS
CAS Number
22569-72-8
7784-34-1
7440-43-9
NA
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 60IOC (SW-846).
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3031 .pdf
4.2.8 EPA Method 3050B (SW-846): Acid Digestion of Sediments, Sludges, and Soils
This method should be used for preparation of solid and oily-solid phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Arsenic (III) compounds
Arsenic Trichloride
CAS Number
22569-72-8
7784-34-1
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Contaminant
Arsine
Cadmium
Metals, NOS
CAS Number
7784-42-1
7440-43-1
NA
A 1 - 2 g sample is digested with nitric acid and hydrogen peroxide. Samples to be analyzed by Method
60IOC (SW-846) for cadmium are also treated with hydrochloric acid. Sample volumes are reduced, then
brought up to a final volume of 100 mL. Samples are analyzed for arsenic by Method 6020A (SW-846)
and for cadmium by either Method 60IOC or 6020A.
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3050b.pdf
4.2.9 EPA Method 3520C (SW-846): Continuous Liquid-Liquid Extraction
This method should be used for preparation of aqueous/liquid phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Bromadiolone
Chloropicrin
Chlorosarin
Chlorosoman
Cyclohexyl Sarin (GF)
Dichlorvos
Dicrotophos
Diesel Range Organics
Dimethylphosphite
Distilled Mustard (HD)/Mustard Gas (H)
Ethyldichloroarsine (ED)
Fenamiphos
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Kerosene
Lewisite 1 (L-1 )
Lewisite 2 (L-2)
Lewisite 3 (L-3)
CAS Number
28772-56-7
76-06-2
1445-76-7
7040-57-5
329-99-7
62-73-7
141-66-2
NA
868-85-9
505-60-2
598-14-1
22224-92-6
1189-87-3
64742-81-0
541-25-3
40334-69-8
40334-70-1
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Contaminant
Methyl parathion
Mevinphos
Nicotine
Nitrogen Mustard (HN-2) [unstable compound]
O-ethyl-S-(2-diisopropylaminoethyl)methyl
phosphonothiolate (VX)
Perfluoroisobutylene (PFIB)
Phencyclidine
Phorate
Sarin (GB)
Semivolatile Organic Compounds, NOS
Soman (GD)
Strychnine
Tabun (GA)
Tear Gas (CS), chlorobenzylidene malonitrile
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Trimethyl phosphite
VE
VG
VM
CAS Number
298-00-0
7786-34-7
54-11-5
51-75-2
50782-69-9
382-21-8
60124-79-0
298-02-2
107-44-8
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
121-45-9
21738-25-0
78-53-5
21770-86-5
This method describes a procedure for isolating organic compounds from aqueous/liquid samples. 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 liter, is
placed into a continuous liquid-liquid extractor, adjusted, if necessary, to a specific pH (see Table 1 in
Method 3520C), 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: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3520c.pdf
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4.2.10 EPA Method 3535A (SW-846): Solid-Phase Extraction
This method should be used for preparation of aqueous/liquid phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Bromadiolone
Chloropicrin
Chlorosarin
Chlorosoman
Cyclohexyl Sarin (GF)
Dichlorvos
Dicrotophos
Diesel Range Organics
Dimethylphosphite
Distilled Mustard (HD)/Mustard Gas (H)
Ethyldichloroarsine (ED)
Fenamiphos
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Kerosene
Lewisite 1 (L-1 )
Lewisite 2 (L-2)
Lewisite 3 (L-3)
Methyl parathion
Mevinphos
Nicotine
Nitrogen Mustard (HN-2) [unstable compound]
O-ethyl-S-(2-diisopropylaminoethyl)methyl
phosphonothiolate (VX)
Perfluoroisobutylene (PFIB)
Phencyclidine
Phorate
Sarin (GB)
CAS Number
28772-56-7
76-06-2
1445-76-7
7040-57-5
329-99-7
62-73-7
141-66-2
NA
868-85-9
505-60-2
598-14-1
22224-92-6
1189-87-3
64742-81-0
541-25-3
40334-69-8
40334-70-1
298-00-0
7786-34-7
54-11-5
51-75-2
50782-69-9
382-21-8
60124-79-0
298-02-2
107-44-8
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Contaminant
Semivolatile Organic Compounds, NOS
Soman (GD)
Strychnine
Tabun (GA)
Tear Gas (CS), chlorobenzylidene malonitrile
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Trimethyl phosphite
VE
VG
VM
CAS Number
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
121-45-9
21738-25-0
78-53-5
21770-86-5
This method describes a procedure for isolating target organic analytes from aqueous/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: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3535a.pdf
4.2.11 EPA Method 3541 (SW-846): Automated Soxhlet Extraction
This method should be used for preparation of solid and oily-solid phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Bromadiolone
Chloropicrin
Chlorosarin
Chlorosoman
Cyclohexyl Sarin (GF)
Dichlorvos
Dicrotophos
Diesel Range Organics
CAS Number
28772-56-7
76-06-2
1445-76-7
7040-57-5
329-99-7
62-73-7
141-66-2
NA
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Contaminant
Dimethylphosphite
Distilled Mustard (HD)/Mustard Gas (H)
Ethyldichloroarsine (ED)
Fenamiphos
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Lewisite 1 (L-1 )
Lewisite 2 (L-2)
Lewisite 3 (L-3)
Methyl parathion
Mevinphos
Nicotine
Nitrogen Mustard (HN-2) [unstable compound]
O-ethyl-S-(2-diisopropylaminoethyl)methyl
phosphonothiolate (VX)
Perfluoroisobutylene (PFIB)
Phencyclidine
Phorate
Sarin (GB)
Semivolatile Organic Compounds, NOS
Soman (GD)
Strychnine
Tabun (GA)
Tear Gas (CS), chlorobenzylidene malonitrile
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Trimethyl phosphite
VE
VG
VM
CAS Number
868-85-9
505-60-2
598-14-1
22224-92-6
1189-87-3
541-25-3
40334-69-8
40334-70-1
298-00-0
7786-34-7
54-11-5
51-75-2
50782-69-9
382-21-8
60124-79-0
298-02-2
107-44-8
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
121-45-9
21738-25-0
78-53-5
21770-86-5
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Approximately 10 g of solid sample is mixed with an equal amount of anhydrous sodium sulfate, placed
in an extraction thimble or between two plugs of glass wool, and after adding the appropriate surrogate
amount, is extracted using an appropriate solvent in an automated Soxhlet extractor. 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 for analysis.
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3540c.pdf
4.2.12 EPA Method 3545A (SW-846) Pressurized Fluid Extraction (PFE)
This method should be used for preparation of solid and oily-solid phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Bromadiolone
Chloropicrin
Chlorosarin
Chlorosoman
Cyclohexyl Sarin (GF)
Dichlorvos
Dicrotophos
Diesel Range Organics
Dimethylphosphite
Distilled Mustard (HD)/Mustard Gas (H)
Ethyldichloroarsine (ED)
Fenamiphos
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Kerosene
Lewisite 1 (L-1 )
Lewisite 2 (L-2)
Lewisite 3 (L-2)
Methyl parathion
Mevinphos
Nicotine
Nitrogen Mustard (HN-2) [unstable compound]
CAS Number
28772-56-7
76-06-2
1445-76-7
7040-57-5
329-99-7
62-73-7
141-66-2
NA
868-85-9
505-60-2
598-14-1
22224-92-6
1189-87-3
64742-81-0
541-25-3
40334-69-8
40334-70-1
298-00-0
7786-34-7
54-11-5
51-75-2
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Contaminant
O-ethyl-S-(2-diisopropylaminoethyl)rnethyl
phosphonothiolate (VX)
Perfluoroisobutylene (PFIB)
Phencyclidine
Phorate
Sarin (GB)
Semivolatile Organic Compounds, NOS
Soman (GD)
Strychnine
Tabun (GA)
Tear Gas (CS), chlorobenzylidene malonitrile
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Trimethyl phosphite
VE
VG
VM
CAS Number
50782-69-9
382-21-8
60124-79-0
298-02-2
107-44-8
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
121-45-9
21738-25-0
78-53-5
21770-86-5
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, as
needed, 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 i-ig/kg of semivolatile organic
compounds, 250 to 2500 i-ig/kg of organophosphorus pesticides, 5 to 250 i-ig/kg of organochlorine
pesticides, 50 to 5000 |ig/kg of chlorinated herbicides, 1 to 1400 |ig/kg of PCBs, and 1 to 2500 ng/kg of
PCDDs/PCDFs.
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3545a.pdf
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4.2.13 EPA Method 3580A (SW-846): Waste Dilution
This method should be used for preparation of oily-solid phase samples for the contaminants identified
below and listed in Appendix A:
Contaminant
Bromadiolone
Chloropicrin
Chlorosarin
Chlorosoman
Cyclohexyl Sarin (GF)
Dichlorvos
Dicrotophos
Diesel Range Organics
Dimethylphosphite
Distilled Mustard (HD)/Mustard Gas (H)
Ethyldichloroarsine (ED)
Fenamiphos
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Kerosene
Lewisite 1 (L-1 )
Lewisite 2 (L-2)
Lewisite 3 (L-3)
Methyl parathion
Mevinphos
Nicotine
Nitrogen Mustard (HN-2)
O-ethyl-S-(2-diisopropylaminoethyl)methyl
phosphonothiolate (VX)
Perfluoroisobutylene (PFIB)
Phencyclidine
Phorate
Sarin (GB)
CAS Number
28772-56-7
76-06-2
1445-76-7
7040-57-5
329-99-7
62-73-7
141-66-2
NA
868-85-9
505-60-2
598-14-1
22224-92-6
1189-87-3
64742-81-0
541-25-3
40334-69-8
40334-70-1
298-00-0
7786-34-7
54-11-5
51-75-2
50782-69-9
382-21-8
60124-79-0
298-02-2
107-44-8
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Contaminant
Semivolatile Organic Compounds, NOS
Soman (GD)
Strychnine
Tabun (GA)
Tear Gas (CS), chlorobenzylidene malonitrile
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Trimethyl phosphite
VE
VG
VM
CAS Number
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
121-45-9
21738-25-0
78-53-5
21770-86-5
This method describes a 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: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/358Oa.pdf
4.2.14 EPA Method 3585 (SW-846): Waste Dilution for Volatile Organics
This method should be used for preparation of oily-solid phase samples for the contaminants identified
below and listed in Appendix A:
Contaminant
Carbon disulfide
Cyanogen chloride
Ethylene Oxide
Gasoline Range Organics
Phosgene
Volatile Organic Compounds, NOS
CAS Number
75-15-0
506-77-4
75-21-8
NA
75-44-5
NA
This method describes a 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 - 10.0 mL with «-hexadecane or other appropriate solvent. Diluted samples are injected into the GC or
GC/MS for analysis.
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Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/3585.pdf
4.2.15 EPA Method 5030C (SW-846): Purge-and-Trap for Aqueous Samples
This method should be used for preparation of aqueous/liquid phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Carbon disulfide
Cyanogen chloride
Ethylene oxide
Gasoline Range Organics
Kerosene
Volatile Organic Compounds, NOS
CAS Number
75-15-0
506-77-4
75-21-8
NA
64742-81-0
NA
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. It also describes the analysis of high
concentration soil and waste sample extracts prepared using Method 5035 (SW-846).
Aqueous/liquid Samples: An inert gas is bubbled through a portion of the aqueous/liquid sample at
ambient temperature, and the volatile components are efficiently 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.
High Concentration Extracts from Method 5035 (SW-846): An aliquot of the extract prepared using
Method 5035 is combined with organic-free reagent water in the purging chamber. It is then analyzed by
purge-and-trap GC or GC/MS following the procedure used for the aqueous/liquid samples.
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/503Oc.pdf
4.2.16 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 phase samples for the contaminants identified below
and listed in Appendix A:
Contaminant
Carbon disulfide
Cyanogen chloride
Ethylene oxide
Gasoline Range Organics
Kerosene
CAS Number
75-15-0
506-77-4
75-21-8
NA
64742-81-0
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Contaminant
Phosgene
Volatile Organic Compounds, NOS
CAS Number
75-44-5
NA
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 i-ig/kg) of VOCs. The method also provides
specific procedures for preparation of samples containing high levels (>200 i-ig/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 automatically added, and the vial heated to 40°C. The
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., 8260B).
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/5035a r 1 .pdf
4.2.17 EPA Method 5050 (SW-846): Bomb Preparation Method for Solid Waste
This method should be used for preparation of solid and aqueous/liquid phase samples for the
contaminant identified below and listed in Appendix A:
Contaminant
Osmium tetraoxide
CAS Number
20816-12-0
A 0.5 g sample is placed in a sample cup, which is placed in a bomb. The sample is combusted with pure
oxygen. The bomb and cup are rinsed, and the rinse collected and analyzed using Method 60IOC (SW-
846).
Source: htto://www.epa.gov/epaoswer/hazwaste/test/pdfs/5050.pdf
4.2.18 EPA Method 601OC (SW-846): Inductively Coupled Plasma - Atomic Emission
Spectrometry
This method should be used for determination of the contaminants identified below and listed in
Appendix A:
Contaminant
Arsenic (III) compounds
Arsenic trichloride
Cadmium
Metals, NOS
Osmium tetraoxide
CAS Number
22569-72-8
7784-34-1
7440-43-9
NA
20816-12-0
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Contaminant
Titanium tetrachloride
CAS Number
7550-45-0
This method determines arsenic (III) compounds and arsenic trichloride as arsenic, osmium tetraoxide as
osmium, and titanium tetrachloride as titanium. Any other metals are determined as the metal. Aqueous
samples (prepared using OW Method 200.2 or SW-846 Method 5050), soil samples (prepared using SW-
846 Methods 3050B or 5050), oily solid samples (prepared using SW-846 Methods 3050B or 3031), and
air filter/particle samples (prepared using Inorganic (IO) Method 3.4) are analyzed by Inductively
Coupled Plasma - Atomic Emission Spectrometry (ICP-AES). Detection limits vary with each analyte.
The analytical range may be extended by sample dilution.
Source: htto://www.epa.gov/epaoswer/hazwaste/test/pdfs/601 Oc.pdf
4.2.19 EPA Method 6020A (SW-846): Inductively Coupled Plasma - Mass Spectrometry
This method should be used for determination of the contaminants identified below and listed in
Appendix A:
Contaminant
Arsenic (III) compounds
Arsenic trichloride
Cadmium
Metals NOS
Titanium tetrachloride
CAS Number
22569-72-8
7784-34-1
7440-43-9
NA
7550-45-0
This method determines arsenic (III) compounds, arsenic trichloride, and titanium tetrachloride as
titanium. Any other metals are determined as the metal. Aqueous samples (prepared using OW Method
200.2 or SW-846 Method 5050), soil samples (prepared using SW-846 Methods 3050B or 5050), oily
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. Detection limits
vary with each analyte. The analytical range may be extended by sample dilution.
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/6020a.pdf
4.2.20 EPA Method 7010 (SW-846): Graphite Furnace Atomic Absorption
Spectrophotometry
This method should be used for determination of the contaminant identified below and listed in Appendix
A:
Contaminant
Arsine
CAS Number
7784-42-1
This method determines arsine in environmental samples. Aqueous samples (prepared using OW Method
200.2) or soil samples (prepared using SW-846 Method 3050B) are analyzed by Graphite Furnace Atomic
Absorption Spectrophotometry (GFAA). A representative aliquot of the sample is placed in the graphite
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tube in the furnace, evaporated to dryness, charred, and atomized. Detection limits vary with each matrix
and instrument used. The analytical range may be extended by sample dilution.
Source: htto://www.epa.gov/epaoswer/hazwaste/test/pdfs/7010.pdf
4.2.21 EPA Method 7470A (SW-846): Mercury in Liquid Wastes (Manual Cold-Vapor
Technique)
This method should be used for preparation and analysis of aqueous/liquid samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Mercury
CAS Number
7439-97-6
A 100-mL aqueous or liquid waste 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: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/7470a.pdf
4.2.22 EPA Method 7471B (SW-846): Mercury in Solid or Semisolid Wastes (Manual Cold-
Vapor Technique)
This method should be used for preparation of solid phase samples for the contaminant identified below
and listed in Appendix A:
Contaminant
Mercury
CAS Number
7439-97-6
A 0.5 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: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/7471b.pdf
4.2.23 EPA Method 8015C (SW-846): Nonhalogenated Organics Using GC/FID
This method should be used for determination of the contaminants identified below and listed in
Appendix A:
Contaminant
Diesel Range Organics
Gasoline Range Organics
Kerosene
CAS Number
NA
NA
64742-81-0
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This method provides gas chromatographic 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, direction 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 ^ig/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: htto://www.epa.gov/epaoswer/hazwaste/test/pdfs/8015c.pdf
4.2.24 EPA Method 8260B (SW-846): Volatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS)
This method should be used for determination of the contaminants identified below and listed in
Appendix A:
Contaminant
Carbon disulfide
Cyanogen chloride
Ethylene oxide
Phosgene
Volatile Organic Compounds, NOS
CAS Number
75-15-0
506-77-4
75-21-8
75-44-5
NA
Volatile compounds are introduced into a gas chromatograph by purge-and-trap or other methods (see
Sec. 1.2 in Method 8260B). The analytes are 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. The column is temperature-programmed to separate the analytes, which are then detected with a
mass spectrometer (MS) interfaced to the gas chromatograph (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) of Method 8260B for an individual analyte is dependent on the instrument as
well as the choice of sample preparation/introduction method. Using standard quadrapole instrumentation
and the purge-and-trap technique, estimated quantitation limits are 5 i-ig/kg (wet weight) for soil/sediment
samples and 5 i-ig/L for ground water (see Table 3 in Method 8260B). 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: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/8260b.pdf
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4.2.25 EPA Method 8270D (SW-846): Semivolatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS)
This method should be used for determination of the contaminants identified below and listed in
Appendix A:
Contaminant
Chloropicrin
Chlorosarin
Chlorosoman
Cyclohexyl Sarin
Dichlorvos
Dicrotophos
Dimethylphosphite
Distilled Mustard (HD)/Mustard Gas (H)
Ethyldichloroarsine (ED)
Fenamiphos
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Lewisite 1 (L-1)
Lewisite 2 (L-2)
Lewisite 3 (L-3)
Methyl parathion
Mevinphos
Nicotine
Nitrogen Mustard (HN-2) [unstable compound]
O-ethyl-S-(2-diisopropylaminoethyl)methyl
phosphonothiolate (VX)
Perfluoroisobutylene (PFIB)
Phencyclidine
Phorate
Sarin (GB)
Semivolatile Organic Compounds, NOS
Soman (GD)
Strychnine
CAS Number
76-06-2
1445-76-7
7040-57-5
329-99-7
62-73-7
141-66-2
868-85-9
505-60-2
598-14-1
22224-92-6
1189-87-3
541-25-3
40334-69-8
40334-70-1
298-00-0
7786-34-7
54-11-5
51-75-2
50782-69-9
382-21-8
60124-79-0
298-02-2
107-44-8
NA
96-64-0
57-24-9
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Contaminant
Tabun
Tear gas (CS), chlorobenzylidene malonitrile
Tetraethyl pyrophosphate
Trimethyl phosphite
VE
VG
VM
CAS Number
77-81-6
2698-41-1
107-49-3
121-45-9
21738-25-0
78-53-5
21770-86-5
Samples are prepared for analysis by gas chromatography/mass spectrometry (GC/MS) 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 gas chromatograph.
Analytes eluted from the capillary column are introduced into the mass spectrometer. The estimated
method detection limits vary with each analyte and range between 10 and 1000 i-ig/L for aqueous/liquid
samples and 660 and 3300 i-ig/kg for soil samples. The analytical range depends on the target analyte(s)
and the instrument used.
Source: htto://www.epa.gov/epaoswer/hazwaste/test/pdfs/8270d.pdf
4.2.26 EPA Method 8315A (SW-846): Determination of Carbonyl Compounds by High
Performance Liquid Chromatography (HPLC)
This method should be used for preparation of solid and aqueous/liquid phase samples for the
contaminants identified below and listed in Appendix A:
Contaminant
Formaldehyde
CAS Number
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 which permit 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 i-ig/L for aqueous/liquid samples.
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/8315a.pdf
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4.2.27 EPA Method 8318A (SW-846): A/-Methylcarbamates by High Performance Liquid
Chromatography (HPLC)
This method should be used for preparation of solid, oily-solid, and aqueous/liquid phase samples for the
contaminants identified below and listed in Appendix A:
Contaminant
Aldicarb (Temik)
Carbofuran (Furadan)
Oxamyl
CAS Number
116-06-3
1563-66-2
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 C-18 cartridge, filtered, and eluted on a C-18 analytical column. After
separation, the target analytes are hydrolyzed and derivatized post-column, then quantitated
fluorometrically. The sensitivity of the method usually depends on the level of interferences present,
rather than on the instrumental 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 and 9.4 i-ig/L for aqueous/liquid samples and 10 and
50 i-ig/kg for soil samples.
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/8318a.pdf
4.2.28 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 determination of the contaminants identified below and listed in
Appendix A:
Contaminant
Bromadiolone
Tetramethylenedisulfotetramine
CAS Number
28772-56-7
80-12-6
This method provides reversed-phase high performance liquid chromatographic (RP/HPLC),
thermospray (TS) 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
TS/MS. Quantitative analysis may be performed by either TS/MS or UV detection, using either an
external or internal standard approach. TS/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 analytical range and detection limits vary depending on the target analyte and instrument used.
Source: http://www.epa.gov/epaoswer/hazwaste/test/pdfs/8321b.pdf
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4.2.29 AOAC Official Method 994.08: Aflatoxin in Corn, Almonds, Brazil Nuts, Peanuts,
and Pistachio Nuts
This method should be used for preparation of solid, oily-solid, and aqueous/liquid phase samples for the
contaminants identified below and listed in Appendix A:
Contaminant
Aflatoxin
Brevetoxins
Picrotoxin
Saxitoxin
T-2 Mycotoxins
CAS Number
1402-68-2
NA
124-87-8
35523-89-8
NA
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 liquid chromatography 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
5-30 ng/g. Note: This method was originally designed for the analysis of aflatoxins in commodities where
cleanup was necessary to remove other food components, like fats and proteins; the cleanup procedure
may not be necessary with water analyses.
Source: AOAC International. 1998. Official Methods of Analysis of AOAC International. 16th Edition, 4th
Revision: Vol II.
4.2.30 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 of solid phase samples for the contaminant identified below
and listed in Appendix A:
Contaminant
Asbestos
CAS Number
1332-21-4
This method describes procedures to (a) identify asbestos in dust and (b) 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. A section of the membrane is prepared and transferred
to a 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.
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Source:
http://www.astm.org/cgi-bin/SoftCart.exe/STORE/filtrexx40.cgi?U+mvstore+tavs3076+-L+D5755:03+/u
sr6/htdocs/astm.org/DATABASE.CART/REDLINE PAGES/05755 .htm
4.2.31 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 of solid phase samples for the contaminant identified below
and listed in Appendix A:
Contaminant
Asbestos
CAS Number
1332-21-4
This test 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. 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 EDXA at a magnification from 15,000 to
20,OOOX.
Source:
httD://www.astm.org/cgi-bin/SoftCart.exe/STORE/filtrexx40.cgi?U+mvstore+tavs3076+-L+D6480:99+/u
sr6/htdocs/astm.org/DATABASE.CART/REDLINE PAGES/D6480.htm
4.2.32 ISO Method -10312: Ambient Air - Determination of Asbestos Fibres -
Direct-transfer Transmission Electron Microscopy Method (TEM)
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Asbestos
CAS Number
1332-21-4
This method determines the type(s) of asbestos fibers present but cannot discriminate between individual
fibres 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 are of 0.195 mm of the
TEM specimen is examined.
Source: http://www.iso.org
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4.2.33 NIOSH Method 6001: Arsine
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Arsine
CAS Number
7784-42-1
In this method, arsine is determined as arsenic. A volume of 0.1 to 10 L of air is drawn through a sorbent
tube containing activated charcoal. The sorbent is extracted with a nitric acid solution. The arsine 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: http://www.cdc.gov/niosh/nmam/pdfs/6001 .pdf
4.2.34 NIOSH Method 6002: Phosphine
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Phosphine
CAS Number
7803-51-2
In this method, phosphine is determined as phosphate. One to 16 liters 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, the phosphate is determined by visible spectrometry. The working range
of the method is 0.02 - 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: http://www.cdc.gov/niosh/nmam/pdfs/6002.pdf
4.2.35 NIOSH Method 6004: Sulfur Dioxide
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Sulfur Dioxide
CAS Number
7446-09-5
In this method, sulfur dioxide is determined as sulfite plus sulfate. A volume of 40 to 200 liters 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 - 20 mg/m3 for a 100-L sample. The method is subject to
interference from sulfur trioxide in dry conditions.
Source: http://www.cdc.gov/niosh/nmam/pdfs/6004.pdf
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4.2.36 NIOSH Method 6010: Hydrogen Cyanide
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Hydrogen Cyanide
CAS Number
74-90-8
Hydrogen cyanide is determined as a cyanide ion complex by this method. A volume of 2 to 90 liters 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. The extract is pH
adjusted and treated with the coupling-color agent. The cyanide ion is determined by visible
spectrophotometry. The working range of the method is 3 - 260 mg/m3 for a 3-L sample. The method is
subject to interference from high concentrations of hydrogen sulfide.
Source: http://www.cdc.gov/niosh/nmam/pdfs/6010 .pdf
4.2.37 NIOSH Method 6011: Bromine and Chlorine
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Chlorine
CAS Number
7782-50-5
In this method, chlorine is determined as chloride. A volume of 2 to 90 liters of air is drawn through a
silver membrane filter. A prefilter is used to remove particulate chlorides. The filter is extracted with
sodium hyposulfate solution, and the extract analyzed for chloride by ion chromatography. The working
range of the method is 0.02 - 1.5 mg/m3 for a 90-L sample. The method is subject to positive interference
by HC1 and negative interference by hydrogen sulfide.
Source: http://www.cdc.gov/niosh/nmam/pdfs/6011 .pdf
4.2.38 NIOSH Method 6013: Hydrogen Sulfide
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Hydrogen Sulfide
CAS Number
7783-06-4
Hydrogen sulfide is determined as sulfate by this method. A volume of 1.2 to 40 liters 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 - 20 mg/m3 for a 20-L sample. The method is
subject to interference from sulfur dioxide.
Source: http://www.cdc.gov/niosh/nmam/pdfs/6013 .pdf
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4.2.39 NIOSH Method 6015: Ammonia
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Ammonia
CAS Number
7664-41-7
Ammonia is determined as indophenol blue by this method. A volume of 0.1 to 96 liters 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 - 300 mg/m3 for a 10-L sample. No interferences have been
identified.
Source: http://www.cdc.gov/niosh/nmam/pdfs/6015 .pdf
4.2.40 NIOSH Method 6402: Phosphorus Trichloride
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Phosphorus Trichloride
CAS Number
7719-12-2
In this method, phosphorus trichloride is determined as phospate. A volume of 11 to 100 liters 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 - 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.
Source: http://www.cdc.gov/niosh/nmam/pdfs/6402.pdf
4.2.41 NIOSH Method 7903: Acids, Inorganic
This method should be used for preparation and analysis of gas phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Hydrogen Bromide
Hydrogen Chloride
Hydrogen Fluoride
CAS Number
10035-10-6
7647-01-0
7664-39-3
Acids are analyzed as bromide, chloride, and fluoride, respectively, by this method. A volume of 3 to 100
liters 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 - 5 mg/m3 for a 50-L sample. Particulate salts of the acids are an interference (trapped
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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: http://www.cdc.gov/niosh/nmam/pdfs/7903.pdf
4.2.42 NIOSH Method 7904: Cyanides, Aerosol and Gas
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Cyanide
CAS Number
57-12-5
In this method, cyanide(s) are determined as cyanide ion. A volume of 10 to 180 liters of air is drawn
through a 0.8-(im PVC membrane filter and a bubbler containing 0. IN KOH solution. The filter collects
aerosols of cyanide solutions and the bubbler collects HCN. The filters are extracted with the KOH
solution. Sulfide must be removed from the solutions prior to analysis. Analyze the solutions by cyanide
ion specific electrode (ISE). The working range of the method is 0.5 - 15 mg/m3 for a 90-L sample.
Sulfide, chloride, iodide, bromide, cadmium, zinc, silver, nickel, cuprous iron, and mercury are
interferences.
Source: http://www.cdc.gov/niosh/nmam/pdfs/7904.pdf
4.2.43 NIOSH Method 7906: Fluorides, Aerosol and Gas
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Hydrogen Fluoride
CAS Number
7664-39-3
Hydrogen fluoride is determined as fluoride ion by this method. A volume of 1 to 800 liters 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 - 8
mg/m3 for 250-L samples. If other aerosols are present, gaseous fluoride may be slightly underestimated
owing to adsorption onto or reaction with particles; with concurrent overestimation of particulate/gaseous
fluoride ratio.
Source: http://www.cdc.gov/niosh/nmam/pdfs/7906.pdf
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4.2.44 NIOSH Method S301-1: Fluoroacetate Anion
This method should be used for preparation and analysis of gas phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Fluoroacetate salts
CAS Number
NA
This method was developed specifically for sodium fluoroacetate, but also may be applicable to other
fluoroacetate salts. A known volume of air, 480 L, 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 - 0.16 mg/m3. The detection limit of
the analytical method is estimated to be 20 ng of sodium fluoroacetate per injection, corresponding to a
100-^L aliquot of a 0.2 ^g/rnL standard.
Source: http://www.cdc.gov/niosh/pdfs/s301 .pdf
4.2.45 OSHA Method ID-188: Ammonia in Workplace Atmospheres - Solid Sorbent
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Ammonia
CAS Number
7664-41-7
In this method, ammonia is determined as ammonium ion. A volume of 7.5 to 24 liters of air is drawn
through a sulfuric acid-treated carbon bead sorbent. The sorbent is extracted with reagent water and the
extract analyzed for ammonium by ion chromatography. The detection limit for the method is 0.60 ppm
for 24-L samples and the quantitation limit is 1.5 ppm for 24-L samples. Volatile amines,
monethanolamine, isopropanolamine, and propanolamine may be interferences. Particulate ammonium
salts can be a positive interference (trapped on the glass wool filter plug in the sorbent tube).
Source: http://www.osha-slc.gov/dts/sltc/methods/inorganic/idl88/idl88.html
4.2.46 OSHA Method ID-216SG: Boron Trifluoride (BF3)
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Boron Trifluoride
CAS Number
7637-07-2
Boron trifluoride is determined as fluoroborate by this method. A volume of 30 to 480 liters 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: http://www.osha-slc.gov/dts/sltc/methods/partial/id216sg/id216sg.html
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4.2.47 Standard Method 4110 B: Ion Chromatography with Chemical Suppression of
Eluent Conductivity
This method should be used for preparation and analysis of aqueous/liquid phase samples for the
contaminants identified below and listed in Appendix A:
Contaminant
Hydrogen Bromide
Hydrogen Chloride
CAS Number
10035-10-6
7647-01-0
The contaminants are determined as bromide and chloride respectively by this method. Aqueous/liquid
samples are pre-filtered and injected onto the ion chromatograph. The anions are identified on the basis of
retention time and quantified by measurement of peak area or height. The method can detect bromide and
chloride at 0.1 mg/L. Lower values can be achieved using a higher scale setting and an electronic
integrator. Other salts of the anions are a positive interference. Low molecular weight organic acids may
interfere with chloride.
Source: American Public Health Association, American Water Works Association, and Water
Environment Federation. 1998. Standard Methods for the Examination of Water and Wastewater. 20th
Edition, (http://www.standardmethods.org/)
4.2.48 Standard Method 4500-NH3 B: Preliminary Distillation Step
This method should be used for preparation of aqueous/liquid phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Ammonia
CAS Number
7664-41-7
A 0.5 - 1 L water sample is dechlorinated, buffered, adjusted to pH 9.5, and distilled into a sulfuric acid
solution. The distillate is brought up to volume and neutralized with sodium hydroxide. The distillate is
analyzed by Method 4500-NH3 G.
Source: American Public Health Association, American Water Works Association, and Water
Environment Federation. 1998. Standard Methods for the Examination of Water and Wastewater. 20th
Edition, (http://www.standardmethods.org/)
4.2.49 Standard Method 4500-NH3 G: Automated Phenate Method
This method should be used for analysis of aqueous/liquid phase samples for the contaminant identified
below and listed in Appendix A:
Contaminant
Ammonia
CAS Number
7664-41-7
Ammonia is determined as indophenol blue by this method. A portion of the neutralized distillate from
procedure 4500-NH3 B is run through the manifold decribed in the method. The ammonium in the
distillate reacts with solutions of disodium EDTA, sodium phenate, sodium hypochlorite, and sodium
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nitroprusside. The resulting indophenol blue is detected by colorimetry in a flow cell. The range of the
method is 0.02 - 2.0 mg/L.
Source: American Public Health Association, American Water Works Association, and Water
Environment Federation. 1998. Standard Methods for the Examination of Water and Wastewater. 20th
Edition, (http://www.standardmethods.org/)
4.2.50 Standard Method 4500-CI G: DPD Colorimetric Method
This method should be used for preparation of aqueous/liquid phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Chlorine
CAS Number
7782-50-5
A portion of aqueous/liquid sample is buffered and reacted with N,N-diethyl-p-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 Water Works Association, and Water
Environment Federation. 1998. Standard Methods for the Examination of Water and Wastewater. 20th
Edition, (http://www.standardmethods.org/)
4.2.51 IO Compendium Method IO-3.1: Selection, Preparation, and Extraction of Filter
Material
This method should be used for preparation and analysis of gas phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Arsenic (III) compounds
Arsenic trichloride
Cadmium
CAS Number
22569-72-8
7784-34-1
7440-43-9
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 hydrochloric/nitric acid mix and microwave or hotplate
heating. The extract is filtered and worked up to 20 mL. The extract is analyzed by compendium methods
IO-3.4orIO-3.5.
Source: http://www.epa.gov/ttn/amtic/files/ambient/inorganic/mthd-3-l.pdf
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4.2.52 IO Compendium Method IO-3.4: Determination of Metals in Ambient Participate
Matter Using Inductively Coupled Plasma (ICP) Spectroscopy
This method should be used for preparation and analysis of gas phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Arsenic (III) compounds
Arsenic Trichloride
Cadmium
Osmium Tetraoxide
CAS Number
22569-72-8
7784-34-1
7440-43-9
20816-12-0
All analytes are determined as the metal by this method. Ambient air is sampled by high-volume filters
using compendium method IO-2.1 (a sampling method). The filters are extracted by compendium method
IO-3.1 and the extracts analyzed by Inductively Couple Plasma - Atomic Emission Spectroscopy (ICP-
AES). Detection limits, ranges, and interference corrections are dependent on the analyte and the
instrument used.
Source: http://www.epa.gov/ttn/amtic/files/ambient/inorganic/mthd-3-4.pdf
4.2.53 IO 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 preparation and analysis of gas phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Arsenic (III) compounds
Arsenic trichloride
Arsine
Cadmium
CAS Number
22569-72-8
7784-34-1
7784-42-1
7440-43-9
All analytes are determined as the metal by this method. Ambient air is sampled by high-volume filters
using compendium method IO-2.1 (a sampling method). The filters are extracted by compendium method
IO-3.1 and the extracts analyzed by Inductively Couple Plasma/Mass Spectrometry (ICP/MS). Detection
limits, ranges, and interference corrections are dependent on the analyte and the instrument used.
Source: http://www.epa.gov/ttn/amtic/files/ambient/inorganic/mthd-3-5.pdf
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4.2.54 IO 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 of analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Mercury
CAS Number
7439-97-6
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 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: http://www.epa.gov/ttn/amtic/files/ambient/inorganic/mthd-5.pdf
4.2.55 EPA Air Toxics Method - 6 (TO-6): Method for the Determination of Phosgene
in Ambient Air Using High Performance Liquid Chromatography
This method should be used for preparation and analysis of gas phase samples for the contaminant
identified below and listed in Appendix A:
Contaminant
Phosgene
CAS Number
75-44-5
This method can be used to detect phosgene in air at the 0.1 ppbv level. Ambient air is drawn through a
midget impinger containing 10 mL of 2/98 aniline/toluene (by volume). Phosgene readily reacts with
aniline to form carbanilide (1,3-diphenylurea), which is stable indefinitely. After sampling, the impinger
contents are transferred to a screw capped vial having a Teflon-lined cap and returned to the laboratory
for analysis. The solution is taken to dryness by heating, and the residue is dissolved acetonitrile.
Carbanilide is determined in the acetonitrile solution using reverse-phase HPLC with an ultraviolet (UV)
absorbance detector operating at 254 nm. Precision for phosgene spiked into a clean air stream is ±15-
20% relative standard deviation. Recovery is quantitative within that precision, down to less than 3 ppbv.
Source: http://www.epa.gov/ttn/amtic/files/ambient/airtox/to-6.pdf
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4.2.56 EPA Air Toxics - 13A (TO-13A): of Polycyclic
(PAHs) in Air
This method should be used for preparation and analysis of gas phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Aldicarb (Temik)
Bromadiolone
Carbofuran (Furadan)
Chloropicrin
Chlorosarin
Chlorosoman
Cyclohexy! Sarin (GF)
Dichlorvos
Dicrotophos
Dimethylphosphite
Distilled Mustard (HD)/Mustard Gas (H)
Ethyldichloroarsine (ED)
Fenamiphos
1-Methylethyl ester ethylphosphonofluoridic acid (GE)
Lewisite 1 (L-1 )
Lewisite 2 (L-2)
Lewisite 3 (L-3)
Methyl parathion
Mevinphos
Nicotine
Nitrogen Mustard (HN-2) [unstable compound]
O-ethyl-S-(2-diisopropylaminoethyl)methyl
phosphonothiolate (VX)
Oxamyl
Perfluoroisobutylene (PFIB)
Phencyclidine
Phorate
Sarin (GB)
CAS Number
116-06-3
28772-56-7
1563-66-2
76-06-2
1445-76-7
7040-57-5
329-99-7
62-73-7
141-66-2
868-85-9
505-60-2
598-14-1
22224-92-6
1 1 89-87-3
541-25-3
40334-69-8
40334-70-1
298-00-0
7786-34-7
54-11-5
51-75-2
50782-69-9
23135-22-0
382-21-8
60124-79-0
298-02-2
107-44-8
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Contaminant
Semivolatile Organic Compounds, NOS
Soman (GD)
Strychnine
Tabun (GA)
Tear Gas (CS), chlorobenzylidene malonitrile
Tetraethyl pyrophosphate
Tetramethylenedisulfotetramine
Trimethyl phosphite
VE
VG
VM
CAS Number
NA
96-64-0
57-24-9
77-81-6
2698-41-1
107-49-3
80-12-6
121-45-9
21738-25-0
78-53-5
21770-86-5
Approximately 300 m3 of air is drawn through the filter and sorbent cartridge using a high-volume flow
rate air sampler or equivalent. The filters and sorbent cartridge are extracted by Soxhlet extraction with
appropriate solvent. The extract is concentrated by Kuderna-Danish (K-D) evaporator, followed by silica
gel cleanup using column chromatography to remove potential interferences prior to analysis by GC/MS.
With optimization to reagent purity and analytical conditions, the detection limits for the GC/MS method
range from 1 ng to 10 pg based on field experience.
Source: http://www.epa.gov/ttn/amtic/files/ambient/airtox/to-13arr.pdf
4.2.57 EPA Air Toxics Method -15 (TO-15): Determination of Volatile Organic
Compounds (VOCs) in Air Collected in Specially-Prepared (Summa) Canisters and
Analyzed by Gas Chromatography/Mass Spectrometry (GC/MS)
This method should be used for preparation and analysis of gas phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Carbon disulfide
Cyanogen chloride
Ethylene oxide
Formaldehyde
Volatile Organic Compounds, NOS
CAS Number
75-15-0
506-77-4
75-21-8
50-00-0
NA
The atmosphere is sampled by introduction of air into a specially prepared stainless steel canister
(summa). A sample of air is drawn through a sampling train comprised of components that regulate the
rate and duration of sampling into the pre-evacuated and passivated summa canister. 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
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from the canister through a solid multisorbent concentrator. After the concentration and drying steps are
completed, the 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 GC/MS. This method applies to
ambient concentrations of VOCs above 0.5 ppbv and typically requires VOC enrichment by concentrating
up to 1-L of a sample volume.
Source: http://www .epa.gov/ttn/amtic/files/ambient/airtox/to-15 r .pdf
4.2.58 Journal of Analytical Atomic Spectrometry, 2000, 15, pp. 277-279: Boron
Trichloride Analysis
This method should be used for preparation and analysis of gas phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Boron trichloride
CAS Number
10294-34-5
An analytical procedure is described for the determination of boron trichloride by ICP-AES. A modified
sampling and gas introduction system allows on-line monitoring of the investigated gas. The gas-phase
sample is introduced into an aqueous mannitol solution, and the boron trichloride hydrolyzed to boric
acid. Since this method is designed for measuring dichlorosilane, the detection limit was found to be 0.63
l^g of boron/g dichlorosilane.
Source: Journal of Analytical Atomic Spectrometry, 2000, 15, pp. 277-279: "On-line monitoring of boron
in dichlorosilan by means of inductively coupled plasma atomic emission Spectrometry," Royal Society of
Chemistry. http://pubs.rsc.org/ei/JA/2000/a908634i.pdf
4.2.59 Analytical Letters, 1994, 27 (14), pp. 2703-2718: Screening-Procedure for Sodium
Fluoracetate (Compound 1080) at Sub-Microgram/Gram Concentrations in Soils
This method should be used for preparation of solid and oily-solid phase samples for the contaminants
identified below and listed in Appendix A:
Contaminant
Fluoroacetate salts
CAS Number
NA
Sodium fluoroacetate (Compound 1080) is readily quantitated at sub-microgram per gram concentrations
in small (ca. 1 g) soil samples. Samples are ultrasonically extracted with water, which is then partitioned
with hexane, and acidified prior to re-extraction with ethyl acetate. The latter is taken to dryness in the
presence of triethanolamine "keeper," and the resulting acid is derivatized with pentafluorobenzyl
bromide. Quantitation is performed using a gas chromatograph equipped with an electron-capture
detector. A standardized statistical protocol is used to validate a screening level of 0.2 |^g Compound
1080/g soil. Difluoroacetic, trifluoroacetic, and naturally occurring formic acids do not interfere with the
determination. The recovery for Compound 1080 was 40% from soil fortified to 0.2 (ig/g soil.
Source: Tomkins, B.A., "Screening-Procedure for Sodium Fluoracetate (Compound 1080) at Sub-
Microgram/Gram Concentrations in Soils" Analytical Letters. 27(14), 2703-2718 (1994).
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Section 5.0: Biological Methods
The purpose of this section is to provide analytical methods for the analysis of environmental samples for
biological agents in response to a homeland security event.
Protocols from peer reviewed journal articles have been identified for analyte-matrix pairs where methods
are not available. It should be noted that the limitations of these protocols are not the same as the
limitations of the standardized methods that have been identified. Future steps include the development of
standardized methods based on the journal protocols. The literature references will be replaced with
standardized, validated protocols as they become available.
Although culture-based methods have been selected for the bacterial pathogens, for viruses, PCR
techniques will be used because of the difficulty and time required to culture viruses. It should be noted
that PCR techniques have limitations such as the inability to determine viability or infectivity of the
analyte.
Sample collection and handling protocols are not available for all matrices included in this document.
Future research will include the development and validation of sampling protocols.
A complete list of recommended biological methods for use in response to homeland security events is
provided in Appendix B. The following information is included in Appendix B, organized by type of
organism (e.g., bacteria, virus, protozoa):
• Analyte(s). The contaminant or contaminant(s) of interest.
Determinative technique. An analytical instrument or technique used to determine the quantity and
identification of compounds or components in a sample.
Determinative method identifier. The unique identifier or number assigned to an analytical method
by the method publisher.
Sample preparation procedure and/or sampling method. The recommended sample preparation
procedure and/or sample collection procedure for the analyte-matrix combination.
5.1 General Guidance
The guidance summarized in this section provides a general overview of how to identify the appropriate
biological method(s) for a given analyte-matrix combination as well as recommendations for quality
control procedures.
For additional information on the properties of the biological agents listed in Appendix B, TOXNET
(http://toxnet.nlm.nih.gov/index.html). a cluster of databases on toxicology, hazardous chemicals, and
related areas maintained by the National Library of Medicine is an excellent resource. Additional
information also can be found on CDC's Emergency Preparedness and Response website
(http://www.bt.cdc.gov/). Further research on biological agents is ongoing within EPA and databases to
manage this information are currently under development.
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5.1.1 Standard Operating Procedures for Identifying Biological Methods
To determine the appropriate method and sample preparation procedure and/or sampling method that is to
be used on the environmental samples, locate the analyte of interest in Appendix B: Biological Methods
under the "Analyte" column.
After locating the analyte of interest, continue across the table to identify the appropriate matrix (e.g.,
water, dust, aerosol). This will identify the determinative technique, determinative method, and sample
preparation procedure and/or sampling method for the analyte of interest.
Sections 5.2.1 through 5.2.15 below provide summaries of the analytical methods listed in Appendix B.
Where available, a direct link to the full text of the selected 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 2.
Table 2. Sources of Biological Methods
Name
National Environmental Methods Index
(NEMI)
U.S. EPA Microbiology Methods
USDA/FSIS Microbiology Laboratory
Guidebook
ICR Microbial Laboratory Manual
Occupational Safety and Health
Administration Methods
National Institutes for Occupational
Safety and Health Methods
Standard Methods for the Examination
of Water and Wastewater, 20th Edition
Annual Book of ASTM Standards
Applied and Environmental
Microbiology
Journal of Clinical Microbiology
Publisher
EPA, USGS
EPA
USDA Food Safety and
Inspection Service
EPA Office of Research and
Development
OSHA
NIOSH
American Public Health
Association and American
Water Works Association
ASTM International
American Society for
Microbiology
American Society for
Microbiology
Reference
http://vwwv.nemi.gov
http://www.epa.qov/microbes/
http://www.fsis.usda.qov/OPHS/microla
b/mlqbook.htm
http://www.epa. qov/nerlcwww/icrmicro.p
d|
http://www.osha-slc.qov/dts/sltc/method
s/toc.html
http://www.cdc.gov/niosh/nmam/
http://www.apha.org
http://www.awwa.org
ISBN: 0875532357
http://www.astm.org
http://www.asm.org
http://www.asm.org
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5.1.2 General Quality Control (QC) Guidance for Biological Methods
Having data of known and documented quality is critical in order for public officials to accurately assess
how to respond to emergency situations. 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.
While method-specific QC requirements are described in many of the individual methods that are cited in
this manual, and will be included in any standardized analytical protocols developed to address specific
analytes and matrices of concern, the following describes a minimum set of QC procedures that shall be
conducted for all biological testing. Individual methods, sampling and analysis protocols, or contractual
statements of work also should be consulted to determine any additional QC that may be needed. These
QC requirements generally consist of analysis of laboratory control samples and or matrix spikes to
identify and quantify measurement system accuracy at the levels of concern, blanks as a measure of
freedom from contamination, and matrix spike duplicates (MSB) or sample replicates to assess data
precision. 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 analysis of standards and other samples to ensure the continued reliability of the analytical
results. Examples of sufficient QC includes:
• Demonstration that measurement system is operating properly
*• Initial calibration (for molecular techniques)
*• Method blanks
• Demonstration of initial and ongoing method performance
*• Precision and recovery
*• Matrix spike/matrix spike duplicates
*• QC samples
*• Method blanks
Please note: The appropriate point of contact identified in Section 3 should be consulted regarding
appropriate quality assurance and quality control procedures prior to sample analysis.
5.1.3 Safety and Waste Management
It is imperative that safety precautions are used during collection, processing, and analysis of
environmental samples, particularly in emergency response situations that may include unknown hazards.
Many of the methods summarized or cited in Section 5.2 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. Additional resources that can be consulted for additional information include the following:
• Environmental Protection Agency's standards regulating hazardous waste (40 CFR parts 260 - 270)
• Biosafety in Microbiological and Biomedical Laboratories, 4th Edition, found at
www. cdc. gov/od/ohs/biosftv/bmb 14toc .htm
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Laboratory Security and Emergency Response Guidance for Laboratories Working with Select
Agents, December 6, 2002 751 (RR19); 1-8, found at
www.cdc.gov/mmwr/preview/mmwrhtml/rr5119al .htm.
5.2 Method Summaries
Method summaries for the analytical methods listed in Appendix B are provided in Sections 5.2.1 -
5.2.15. Each method summary contains a table identifying the contaminants in Appendix B to which the
method applies, provides a brief description of the analytical method, and includes a link to the full
version of the method or source for obtaining a full version of the method. Any questions regarding
information discussed in this section should be addressed to the appropriate contact(s) listed in Section 3.
5.2.1 Laboratory Response Network (LRN)
The agents identified below and listed in Appendix B should be analyzed in accordance with the
appropriate LRN protocols:
Analyte(s)
Bacillus anthracis (Anthrax)
Brucella spp. (Brucellosis)
Burkholderia mallei (Glanders)
Burkholderia pseudomallei (Melioidosis)
Francisella tularensis (Tularemia)
Coxiella burnetti (Q-fever)
Yersina pestis (Plague)
Variola major (Smallpox)
Agent Category
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Viruses
These agents will be analyzed using restricted procedures available only through the Laboratory Response
Network (LRN). These procedures are not available to the general laboratory community and thus are not
discussed within this document. For additional information on the LRN, please use the contact
information provided below or visit http://www.bt.cdc.gov/lrn/.
Centers for Disease Control and Prevention
Laboratory Response Branch
Bioterrorism Preparedness and Response Program
National Center for Infectious Diseases
1600 Clifton Road NE, Mailstop C-18
Atlanta, GA 30333
Telephone: (404) 639-2790
E-mail: lrn@,cdc.gov
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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
2025 M Street NW, Suite 550
Washington, DC 20036
Telephone: (202) 822-5227
Fax: (202) 887-5098
Website: www.aphl.org
E-mail: info@aplil.org
5.2.2 Biosafety Level 4 Viruses
Samples to be analyzed for the viruses identified below and listed in Appendix B should be analyzed
under biosafety level (BSL) 4 conditions at the Centers for Disease Control and Prevention (CDC):
Analyte(s)
Arenaviruses
Nairovirus
Hemorrhagic fever viruses
Rift vally fever
Variola major (Smallpox)
Agent Cateory
Viruses
Viruses
Viruses
Viruses
Viruses
For additional information on the LRN, please use the contact information provided below or visit
http://www.bt.cdc.gov/lrn/.
Centers for Disease Control and Prevention
Laboratory Response Branch
Bioterrorism Preparedness and Response Program
National Center for Infectious Diseases
1600 Clifton Road NE, Mailstop C-18
Atlanta, GA 30333
Telephone: (404) 639-2790
E-mail: lrn@cdc.gov
5.2.3 U.S. EPA Method 1622: Cryptosporidium in Water by Filtration/IMS/FA
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Cryptosporidium parvum (Cryptosporidiosis)
Agent Category
Protozoa
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
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ax&i-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 (DIG) 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.
Please note: This method was originally developed for water matrices; further research will be conducted
to develop and standardize sample processing protocols for other matrices.
Source: USEPA. 2001. Cryptosporidium in Water by Filtration/IMS/FA (EPA-821-R-01-026). United
States Environmental Protection Agency, Washington, B.C.
(http: //www. epa.gov/nerlcwww/ 1622ap01.pdf)
5.2.4 ICR Microbial Laboratory Manual Chapter XI
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Clostridium perfringens
Agent Category
Bacteria
An appropriate volume of water sample is passed through a membrane filter that retains the bacteria
present in the sample. The membrane filter is placed on mCP agar and incubated anaerobically for 24 h at
44.5° C. Upon exposure to ammonium hydroxide, the yellow straw-colored C. perfringens colonies turn
dark pink to magenta and are counted as presumptive C. perfringens. Biochemical confirmation includes
anaerobic growth in thioglycollate, a positive gram stain reaction and stormy fermentation of iron milk.
Please note: This procedure has not been fully validated. At a minimum, the following sample processing
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. This method was originally developed for water matrices; further research
will be conducted to develop and standardize sample processing protocols for other matrices.
Source: United States Environmental Protection Agency, Office of Research and Development.
1996. ICR Microbial Laboratory Manual Chapter XI. EPA/600/R-95/178.
(http://www.epa.gov/microbes/icrmicro.pdf)
5.2.5 USDA/FSIS 4
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Salmonella typhi (Typhoid fever)
Agent Category
Bacteria
Samples are added to buffered peptone water and incubated for 24 hours. An aliquot of the broth is added
to both tetrathionate broth and modified Rappaport-Vassiliadis broth and incubated for 24 hours. An
aliquot of each broth is streaked onto Brilliant green sulfa agar, Xylose lysine Tergitol™ 4 agar , or
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Double modified lysine iron agar and incubated for 24 hours. Isolates are inoculated into triple sugar iron
and lysine iron agar slants. Additional biochemical tests are performed using commercially available
biochemical test kits. Serological testing is done using polyvalent O antisera. The method addendum (4A)
allows for the use of a rapid screening immunoassay kit.
Please note: This procedure has not been fully validated. At a minimum, the following sample processing
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: Isolation and Identification of Salmonella from Meat, Poultry, and Egg Products. 3rd Edition
1998. http://www.fsis.usda.gov/Qphs/Microlab/Mlg4.02.pdf:
http://www.fsis.usda.gov/Ophs/Microlab/Mlg4CO 1 .pdf
5.2.6 Standard Methods 9260 E: Shigella spp.
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Shigella spp.
Agent Category
Bacteria
This method contains two options for sample concentration: membrane filtration (liquid matrices) and
centrifugation (liquid and solid matrices) for analyses. Both options include inoculation of an enrichment
medium (Selenite F broth). Isolation of the target analyte is achieved by plating onto XLD and/or
MacConkey agar. Biochemical identification consists of inoculating TSI and LIA slants. Serological
grouping is done by slide agglutination tests using polyvalent and group specific antisera.
Please note: This procedure has not been fully validated. At a minimum, the following sample processing
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank.
Source: American Public Health Association, American Water Works Association, and Water
Environment Federation. 1998. Standard Methods for the Examination of Water and Wastewater. 20th
Edition, (http://www.standardmethods.org/)
5.2.7 Standard Methods 9260 F: Pathogenic Escherichia coli
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Escherichia coli (E. coli) O157:H7
Agent Category
Bacteria
This method allows for two options, one being a modification of SM 922 IB 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 EHEC Enterohemorrhagic
E. coli enrichment broth (EEB) and incubated for 6 hours. Tellurite Cefixime SMAC (TC SMAC) plates
are inoculated with the EEB. Both the TC SMAC and EEB are incubated for up to 24 hours. Colorless
colonies on TC SMAC are tested for indole production. Additional biochemical tests and serotyping are
also done to confirm identification.
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Please note: This procedure has not been fully validated. At a minimum, the following sample processing
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. This method was originally developed for liquid matrices; further research
will be conducted to develop and standardize sample processing protocols for other matrices.
Source: American Public Health Association, American Water Works Association, and Water
Environment Federation. 1998. Standard Methods for the Examination of Water and Wastewater. 20th
Edition, (http://www.standardmethods.org/)
5.2.8 Standard Methods 9260 G: Campylobacterjejuni
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Campylobacterjejuni
Agent Category
Bacteria
Water samples are filtered using a cellulose nitrate membrane filter. Filters are placed face down on either
Skirrow's medium or Campy-BAP and incubated for 24 hours. Filters are then transferred to another
selective medium face-down and incubated for a total of 5 days at 42°C under microaerophilic conditions.
Identification is made by culture examination, microscopy, motility test, and biochemical testing.
Biochemical tests include oxidase, catalase, nitrite and nitrate reduction, H2S production, and hippurate
hydrolysis. Serotyping is done using commercially available rapid test kits.
Please note: This procedure has not been fully validated. At a minimum, the following sample processing
quality control checks should be performed and evaluated before using this protocol: positive control,
negative control, and blank. This method was originally developed for water matrices; further research
will be conducted to develop and standardize sample processing protocols for other matrices.
Source: American Public Health Association, American Water Works Association, and Water
Environment Federation. 1998. Standard Methods for the Examination of Water and Wastewater. 20th
Edition, (http://www.standardmethods.org/)
5.2.9 Standard Methods 9260 H: Vibrio cholerae
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Vibrio cholerae (cholera)
Agent Category
Bacteria
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. Vibrio
cholerae isolates are plated on tryptic soy agar with 0.5% NaCl. Biochemical confirmation is done using
multiple tests including but not limited to ONPG, Indole, and Voges-Proskauer. Slide agglutination
assays are used for serological identification.
Please note: This procedure has not been fully validated. At a minimum, the following sample processing
quality control checks should be performed and evaluated before using this protocol: positive control,
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negative control, and blank. This method was originally developed for water matrices, further research
will be conducted to develop and standardize sample processing protocols for other matrices.
Source: American Public Health Association, American Water Works Association, and Water
Environment Federation. 1998. Standard Methods for the Examination of Water and Wastewater. 20th
Edition, (http://www.standardmethods.org/)
5.2.10 Literature Reference for Caliciviruses
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Caliciviruses
Agent Category
Viruses
This molecular detection method is used for the concentration, extraction, and RT-PCR analysis of human
Noroviruses in water and clinical (stool) samples. Water samples are collected by filtration (1MDS filter)
and viruses are eluted using a beef extract solution (1.5%, pH 9.0). Viruses are concentrated using celite
adsorption (pH 4.0), filtration, and 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 viruses are used either directly or following RNA extraction (GITC)
for two-step (RT followed by PCR) RT-PCR analysis using gene-specific (norovirus pol gene) primer
sets. Detection of amplicons is by agarose gel electrophoresis and subsequent confirmation (genotyping)
by sequence analysis of cloned amplicons.
Please note: This procedure has not been fully validated. At a minimum, the following sample processing
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 the current
version of the EPA Quality Assurance/Quality Control Guidance for Laboratories Performing PCR
Analyses on Environmental Samples. Please call the point of contact identified in Section 3 for additional
information. This method was originally developed for water matrices; further research will be conducted
to develop and standardize sample processing protocols for other matrices.
Source: Parshionikar, S. U., Willian-True, S., Fout, G. S., Robbins, D. E., Seys, S. A., Cassady, J. D.,
and Harris, R. 2003. Waterborne Outbreak of 'Gastroenteritis Associated with a Norovirus. Applied and
Environmental Microbiology 69(9): 5263-5268. http://aem.asm.Org/cgi/reprint/69/9/5263
5.2.11 Literature Reference for Enteroviruses
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Enteroviruses
Agent Category
Viruses
This molecular detection method is used for the simultaneous detection of several human enteric viruses
(enteroviruses, reoviruses, and rotaviruses) in groundwater using a multiplex RT-PCR approach. Water
samples are collected by filtration (1MDS filters); 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 elution with
sodium phosphate (0.15 M, pH 9.0), followed by further concentration and processing to remove
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inhibitors (ultracentrifugation, solvent extraction, and MW-exclusion filtration). Concentrated samples
are analyzed by a two-step multiplex RT-PCR (RT followed by PCR) using virus-specific primer sets.
Detection of amplicons is by gel electrophoresis and subsequent confirmation by hybridization (dot-blot)
using digoxigenin-labeled internal (nested) probes.
Please note: This procedure has not been fully validated. At a minimum, the following sample processing
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 or call the point of contact identified in Section 3. This method was
originally developed for water matrices, further research will be conducted to develop and standardize
sample processing protocols for other matrices.
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 Vol. 69 No. 6: 3158-3164.
(http://aem.asm.Org/cgi/reprint/69/6/3158 .pdf)
5.2.12 Literature Reference for Hepatitis A Viruses
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Hepatitis A Viruses
Agent Category
Viruses
This method is used to detect Hepatitis A virus in groundwater samples. It is a multiplex RT-PCR
procedure optimized for hepatitis A viruses. Water samples are collected by filtration (1MDS filters) 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 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 by a two-step multiplex RT-PCR (RT followed
by PCR) using two hepatitis A-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: This procedure has not been fully validated. At a minimum, the following sample processing
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 or call the point of contact identified in Section 3. This method was
originally developed for water matrices; further research will be conducted to develop and standardize
sample processing protocols for other matrices.
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/69/6/3158.pdf)
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5.2.13 Literature Reference for Venezeulan Equine Encephalitis (VEE) Virus
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Venezuelan Equine Encephalitis (VEE) Virus
Agent Category
Viruses
The VEE-specific RT-PCR assay is applied to human sera viruses that are isolated in either Vero cells,
C6/36 cells, or newborn mice. VEE is identified using an indirect immunofluorescence assay. QIAmp
viral RNA kit (Qiagen) is used to extract RNA without the use of TaqExtender and amplification is done
using gene-specific RT-PCR-seminested PCR. Annealing temperature is 55°C in the thermocycler.
Please note: This procedure has not been fully validated for matrices other than human sera. At a
minimum, the following sample processing 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 or call the point of contact
identified in Section 3. This method was originally developed for clinical matrices, further research will
be conducted to develop and standardize sample processing protocols for other matrices.
Source: Linssen, B., Kinney, R. M., Aguilar, P., Russell, K. L., Watts, D. M., Kaaden, O., and Pfeffer M.
2000. Development of Reverse Transcription-PCR Assays Specific for Detection of Equine Encephalitis
Viruses. Journal of Clinical Microbiology. 38(4): 1527-1535.
(http ://j cm. asm. org/cgi/reprint/3 8/4715 27 .pdf)
5.2.14 Literature Reference for Toxoplasma gondii
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Toxoplasma gondii (Toxoplasmosis)
Agent Category
Protozoa
This method uses PCR amplification of T. gondii oocyst DNA using gene-specific (Bl gene) primers (one
primer end - labeled with biotin) and subsequent detection and confirmation by a sandwich hybridization
capture assay and Enzyme Immunoassay (EIA). Amplicons are generated by PCR and are then denatured
and hybridized with FITC-labeled internal (nested) oligoprobes. Following hybridization, amplicons are
bound to avidin-coated (96-well) plates and reacted with anti-FITC monoclonal antibody conjugated to
horseradish peroxidase. Following substrate reaction, hybridization products are detected colorimetrically
(OD 450).
Please note: This procedure has not been fully validated. At a minimum, the following sample processing
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 or call the point of contact identified in Section 3.
Source: Schwab, K. J. and McDevitt, J. J. 2003. Development of a PCR-Enzyme Immunoassay
Oligoprobe Detection Method for Toxoplasma gondii Oocysts, Incorporating PCR Controls. Applied and
Environmental Microbiology. 69(10):5819-5825.
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5.2.15 Entamoeba histolytica: Filtration/IMS/PCR
This method should be used for the contaminant identified below and listed in Appendix B:
Analyte(s)
Entamoeba histolytica
Agent Category
Protozoa
A standardized method/protocol for the analysis of Entamoeba histolytica has not been identified. Further
research will be conducted prior to the next revision of this document to identify an appropriate
method/protocol. In the interim, PCRtest kits may be obtained for analysis of this organism. Please
contact the appropriate point of contact identified in Section 3 if the need to analyze for Entamoeba
histolytica arises.
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Section 6.0: Conclusions
Methods listed in Appendix A (chemical methods) and Appendix B (biological methods) should be used
to assess the extent of contamination and the effectiveness of decontamination in response to a homeland
security event.
As stated in the introduction, the primary objective of this effort was not necessarily to identify the "best"
method, but rather to have a balanced approach between leveraging existing determinative techniques and
methodologies and providing consistent analytical results. The method selected for each analyte matrix
pair was the most general, appropriate, and broadly applicable of available methods. This is a living
document and recommended methods are subject to change based on advances in technology.
Any questions concerning the information in this document should be directed to the appropriate point(s)
of contact identified in Section 3.
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Abbreviations and Acronyms
1 MDS #1 Mark D. Sobsey filter
AEM Applied and Environmental Microbiology
AGI sampler All Glass Impinger Sampler
AOAC AOAC International
ASTM ASTM International
BSL Biosafety Level
°C Degrees Celsius
Campy-BAC Campylobacter-Brucella agar base with sheep blood and antibiotics
CDC Centers for Disease Control and Prevention
CFR Code of Federal Regulations
CLP Contract Laboratory Program
CVAA Cold Vapor Atomic Absorption
CVAFS Cold Vapor Atomic Fluorescence Spectrometry
DAPI 4',6-diamidino-2-phenylindole
DHS Department of Homeland Security
DIC Differential Interference Contrast
DNA Deoxyribonucleic Acid
DNPH 2,4-dinitrophenylhydrazine
DOD Department of Defense
ED Electron Diffraction
EDTA Ethylenediaminetetraacetic acid
EDXA Energy Dispersive X-ray Analysis
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EEB EHEC Enrichment Broth
EHEC Enterohemorrhagic Escherichia coli
EIA Enzyme Immunoassay
ELISA Enzyme-Linked Immunosorbent Assay
EMMI Environmental Monitoring Methods Index
EPA U.S. Environmental Protection Agency
EQL Estimated Quantitation Limit
FA Fluorescence Assay
FBI Forensic Bureau of Investigation
FDA Food and Drug Administration
FID Flame lonization Detector
FITC Fluorescein isothiocyanate
FSIS Food Safety and Inspection Service
GC Gas Chromatograph or Gas Chromatography
GC/MS Gas Chromatograph/Mass Spectrometer or Gas Chromatography/Mass Spectrometry
GFAA Graphite Furnace Atomic Absorption Spectrophotometer or Graphite Furnace Atomic
Absorption Spectrophotometry
GITC Guanidinium iso-thiocyanate
HAV Hepatitis A Virus
HPLC High Performance Liquid Chromatograph or High Performance Liquid Chromatography
1C Ion Chromatograph or Ion Chromatography
ICP Inductively Coupled Plasma
ICP-AES Inductively Coupled Plasma - Atomic Emission Spectrometry
ICR Information Collection Rule
IMS Immunomagnetic Separation
IO Inorganic
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ISO International Organization for Standardization
ISE Ion Specific Electrode
K-D Kuderna-Danish
LIA Lysine Iron Agar
LRN Laboratory Response Network
LSE Liquid/Solid Extraction
MCP Membrane Clostridium perfringens
MS Mass Spectrometer or Mass Spectrometry
MSB Matrix Spike Duplicate
MW Molecular Weight
NA Not Applicable
NEMI National Environmental Methods Index
NERL-CI National Exposure Risk Laboratory-Cincinnati
NHSRC National Homeland Security Research Center
NIOSH National Institute for Occupational Safety and Health
NOS Not Otherwise Specified
NTIS National Technical Information Service
ONPG Ortho-nitrophenyl-p-D-galactopyranoside
OSHA Occupational Safety and Health Administration
OW Office of Water
PAHs Polycyclic Aromatic Hydrocarbons
PCBs Polychlorinated biphenyls
PCDDs Polychlorinated dibenzo-p-dioxins
PCDFs Polychlorinated dibenzofurans
PCR Polymerase Chain Reaction
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PFE Pressurized Fluid Extraction
QC Quality Control
RNA Ribonucleic Acid
RP/HPLC Reversed-Phase High Performance Liquid Chromatography
RT-PCR Reverse Transcription-Polymerase Chain Reaction
SAED Selected Area Electron Diffraction
SM Standard Methods for the Examination of Water and Wastewater
SPE Solid-Phase Extraction
SW Solid Waste
TCBS Thiosulfate Citrate Bile Salts Sucrose
TC SMAC Tellurite Cefixime Sorbitol MaConkey agar
TCLP Toxicity Characteristic Leaching Procedure
TEM Transmission Electron Microscope or Microscopy
TOXNET National Library of Medicine, Toxicological Database
TS Thermospray
TSI Triple Sugar Iron
USDA U.S. Department of Agriculture
USGS U.S. Geological Survey
UV Ultraviolet
VEE Venezeulan Equine Encephalitis
VOCs Volatile Organic Compounds
VOA Volatile Organic Analysis
XLD Xylose lysine desoxycholate
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Appendix A: Chemical
Analyte(s)
Aflatoxin
Aldicarb (Temik)
Alpha amanitin
Ammonia
Arsenic 111 compound,
Cadmium and other metals
Arsenic trichloride (analyze for
Arsenic)
Arsine
Asbestos
Boron trichloride
Boron trifluoride
Botulinum toxin
CAS No.
1402-68-2
116-06-3
NA
7664-41-7
22569-72-8
7784-34-1
7784-42-1
1332-21-4
10294-34-5
7637-07-2
NA
Determinative
Technique
HPLC-FL
HPLC-FL
Immunoassay
Spectrophotometry
ICP-MS/ICP-AES
ICP-MS/ICP-AES
GFAA
TEM
ICP-AES
ISE
Immunoassay
Determinative Method
Identifier
994.08 (AOAC)
8318A(SW-846)
LRN
4500- NH3 G (SM)
6020A/6010C
(SW-846)
6020A/6010C
(SW-846)
7010 (SW-846)
ASTM(dust)/ISO-
1031 2 (air)
Journal Article: J. Anal.
At. Spectrom., 2000, 15,
277-279
ID-216SG(OSHA)
LRN
Solid Prep
Procedure
994.08 (AOAC)
831 8A (SW-846)
LRN
Not of concern in this matrix
(SW-846)
3050B (SW-846)
(SW-846)
ASTM D5755-03 (soft
surfaces-microvac) or D6480-
99 (hard surfaces-wipes)
Not of concern in this matrix
Not of concern in this matrix
LRN
Oily Solid
Procedure
(AOAC)
831 8A (SW-846)
LRN
Not of concern in this
matrix
3031/3050B
(SW-846)
3031/3050B
(SW-846)
Not of concern in this
matrix
Not of concern in this
matrix
Not of concern in this
matrix
Not of concern in this
matrix
LRN
Aqueous and Liquid
Sample Prep Procedure
994.08 (AOAC)
831 8A (SW-846)
LRN
4500-NH3 B (SM)
200.2 (OW)
200.2 (OW)
200.2 (OW)
Not of concern in this matrix
Not of concern in this matrix
Not of concern in this matrix
LRN
Phase Sample Prep
Procedure
Not of concern in this
matrix
TO-13A
LRN
6015(NIOSH)/ID-188
(OSHA)
IO-3.5/IO-3.4/IO-3.1
(ORD)
IO-3.5/IO-3.4/IO-3.1
(ORD)
6001 (NIOSH)
ISO-10312(filter)
Journal Article: J. Anal. At.
Spectrom., 2000, 15, 277-
279
ID-216SG(OSHA)
LRN
Revision 1.0
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Appendix A: Chemical
Analyte(s)
Brevetoxins
Bromadiolone
Cadmium
Carbofuran (Furadan)
Carbon disulfide
Chlorine
Chloropicrin
Chlorosarin
Chlorosoman
Cyanide
Cyanogen chloride
CAS No.
NA
28772-56-7
7440-43-9
1563-66-2
75-15-0
7782-50-5
76-06-2
1445-76-7
7040-57-5
57-12-5
506-77-4
Determinative
Technique
HPLC-MS
HPLC-UV
ICP-MS/ICP-AES
HPLC
GC/MS
1C
GC/MS
GC/MS
GC/MS
Spectrophotometry
GC/MS
Determinative Method
Identifier
994.08 (AOAC)
8321 B (SW-846)
6020A/6010C
(SW-846)
831 8A (SW-846)
(SW-846)
6011 (NIOSH)
(SW-846)
8270D (SW-846)
(SW-846)
CLP ILM05.3 CN
(SW-846)
Solid Prep
Procedure
994.08 (AOAC)
3545A/3541
(SW-846)
(SW-846)
831 8A (SW-846)
(SW-846)
Not of concern in this matrix
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
CLP ILM05.3 CN
(SW-846)
Oily Solid
Procedure
(AOAC)
3545A/3541/3580A
(SW-846)
3031/3050B
(SW-846)
831 8A (SW-846)
3585 (SW-846)
Not of concern in this
matrix
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
Not of concern in this
matrix
3585 (SW-846)
Aqueous and Liquid
Sample Prep Procedure
994.08 (AOAC)
3520C/3535A
(SW-846)
200.2 (OW)
831 8A (SW-846)
5030C (SW-846)
4500-CI G
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
CLP ILM05.3 CN
(SW-846)
Phase Sample Prep
Procedure
Not of concern in this
matrix
TO-13A
IO-3.5/IO-3.4/IO-3.1
(ORD)
TO- 13A/ Method 74
(OSHA)
TO-15
6011 (NIOSH)
TO-13A
TO-13A
TO-13A
7904 (NiOSH)
TO-15
Revision 1.0
A-2
September 29, 2004
-------
Appendix A: Chemical
Analyte(s)
Cyclohexyl Sarin (GF)
Dichlorvos
Dicrotophos
Diesel Range Organics
Dimethylphosphite
Distilled Mustard (HD) /
Mustard Gas (H)
Ethyldichloroarsine (ED)
Ethylene oxide
Fenamiphos
Fiuoroacetate salts
CAS No.
329-99-7
62-73-7
141-66-2
NA
868-85-9
505-60-2
598-14-1
75-21-8
22224-92-6
NA
Determinative
Technique
GC/MS
GC/MS
GC/MS
GC/FID
GC/MS
GC/MS
GC/MS
GC/MS
GC/MS
Ion Chromatography/
GC/ECD
Determinative Method
Identifier
(SW-846)
8270D (SW-846)
(SW-846)
801 5C (SW-846)
8270D (SW-846)
8270D (SW-846)
8270D (SW-846)
8260B (SW-846)
8270D (SW-846)
300.1 (OW)
Solid Prep
Procedure
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545 A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
5035A (SW-846)
3545A/3541
(SW-846)
Journal Article: Analytical
Letters, 1994, 27(14), 2703-
2718
Oily Solid
Procedure
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3585 (SW-846)
3545A/3541/3580A
(SW-846)
Journal Article: Analytical
Letters, 1994, 27(14),
2703-2718
Aqueous and Liquid
Sample Prep Procedure
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
Note: For liquid matrices use
(SW-846)
5030C (SW-846)
3520C/3535A
(SW-846)
300.1 (OW)
Phase Sample Prep
Procedure
TO-13A
TO-13A
TO-13A
Not of concern in this
matrix
TO-13A
TO-13A
TO-13A
TO-15
TO-13A
S301-1 (NIOSH)
Revision 1.0
A-3
September 29, 2004
-------
Appendix A: Chemical
Analyte(s)
Formaldehyde
Gasoline Range Organics
1 -methylethyl ester ethyl-
phosphonofluoridic acid (GE)
Hydrogen bromide
Hydrogen chloride
Hydrogen cyanide
Hydrogen fluoride
Hydrogen sulfide
Kerosene
Lewisite 1 (L-1)
CAS No.
50-00-0
NA
1189-87-3
10035-10-6
7647-01-0
74-90-8
7664-39-3
7783-06-4
64742-81-0
541-25-3
Determinative
Technique
HPLC
GC/FiD
GC/MS
1C
1C
Spectrophotometry /
ISE
1C
1C
GC/FID
GC/MS
Determinative Method
Identifier
8315A(SW-846)
8015C(SW-846)
(SW-846)
4110B(SM)/7903
(NIOSH)
4110B(SM)/7903
(NIOSH)
6010 (NIOSH)
7906 or 7903 (NIOSH)
6013 (NIOSH)
801 5C (SW-846)
8270D (SW-846)
Solid Prep
Procedure
831 5A (SW-846)
5035A (SW-846)
3545A/3541
(SW-846)
Not of concern in this matrix
Not of concern in this matrix
Not of concern in this matrix
Not of concern in this matrix
Not of concern in this matrix
(SW-846)
3545A/3541
(SW-846)
Oily Solid
Procedure
Not of concern in this
matrix
3585 (SW-846)
3545A/3541/3580A
(SW-846)
Not of concern in this
matrix
Not of concern in this
matrix
Not of concern in this
matrix
Not of concern in this
matrix
Not of concern in this
matrix
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
Aqueous and Liquid
Sample Prep Procedure
831 5A (SW-846)
5030C (SW-846)
3520C/3535A
(SW-846)
4110B(SM)
4110B(SM)
Not of concern in this matrix
Not of concern in this matrix
Not of concern in this matrix
(SW-846)
Note: For liquid matrices use
(SW-846)
3520C/3535A
(SW-846)
Phase Sample Prep
Procedure
TO-15
Not of concern in this
matrix
TO-13A
7903 (NIOSH)
7903 (NIOSH)
6010 (NIOSH)
7906 / 7903 (NIOSH)
601 3 (NIOSH)
Not of concern in this
matrix
TO-13A
Revision 1.0
A-4
September 29, 2004
-------
Appendix A: Chemical
Analyte(s)
Lewisite 2 (L-2)
Lewisite 3 (L-3)
Mercury
Metals, NOS
Methyl parathion
Mevinphos
Microcystin
Nicotine
Nitrogen Mustard (HN-2)
[unstable compound]
O-ethyl-S-(2-
diisopropylaminoethyi)methyl
phosphorite (VX)
Osmium tetraoxide (analyze for
Osmium)
CAS No.
40334-69-8
40334-70-1
7439-97-6
NA
298-00-0
7786-34-7
NA
54-11-5
51-75-2
50782-69-9
20816-12-0
Determinative
Technique
GC/MS
GC/MS
CVAA
ICP-MS/ICP-AES
GC/MS
GC/MS
Immunoassay
GC/MS
GC/MS
GC/MS
ICP-AES
Determinative Method
Identifier
(SW-846)
8270D (SW-846)
7471 B (s) / 7470A (aq)
6020A/6010C
(SW-846)
(SW-846)
8270D (SW-846)
LRN
8270D (SW-846)
8270D (SW-846)
8270D (SW-846)
6010C(SW846)
Solid Prep
Procedure
3545A/3541
(SW-846)
3545A/3541
(SW-846)
7471 B
3050B (SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
LRN
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
5050 (SW-846)
Oily Solid
Procedure
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
Not of concern in this
matrix
3031/3050B
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
LRN
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
Not of concern in this
matrix
Aqueous and Liquid
Sample Prep Procedure
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
7470A
200.2 (OW)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
LRN
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
5050 (SW-846)
Phase Sample Prep
Procedure
TO-13A
TO-13A
IO-5
See specific metals
methods
TO-13A
TO-13A
LRN
TO-13A
TO-13A
TO-13A
IO-3.4 (ORD)
Revision 1.0
A-5
September 29, 2004
-------
Appendix A: Chemical
Analyte(s)
Oxamyl
Paraquat
Perfluoroisobutylene (PFIB)
Phencyciidine
Phorate
Phosgene
Phosphine
Phosphorus trichloride
Picrotoxin
Red Phosphorous (RP)
(analyze for total phosphorous)
Ricin
CAS No.
23135-22-0
4685-14-7
382-21-8
60124-79-0
298-02-2
75-44-5
7803-51-2
7719-12-2
124-87-8
7723-14-0
9009-86-3
Determinative
Technique
HPLC
HPLC-UV
GC/MS
GC/MS
GC/MS
GC/MS
UV-VIS
Spectrophotometry
HPLC-MS
Spectrophotometry
(colorimetric)
Immunoassay
Determinative Method
Identifier
8318A(SW-846)
549.2 (OW)
(SW-846)
8270D (SW-846)
(SW-846)
8260B (SW-846)
6002 (NIOSH)
6402 (NIOSH)
994.08 (AC AC)
365.1 (NERL)
LRN
Solid Prep
Procedure
831 8A (SW-846)
Problematic
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
5035A (SW-846)
Not of concern in this matrix
Not of concern in this matrix
(AOAC)
Not of concern in this matrix
LRN
Oily Solid
Procedure
831 8A (SW-846)
Problematic
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
(SW-846)
3585 (SW-846)
Not of concern in this
matrix
Not of concern in this
matrix
994.08 (AOAC)
Not of concern in this
matrix
LRN
Aqueous and Liquid
Sample Prep Procedure
831 8A (SW-846)
549.2 (OW)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
Not of concern in this matrix
Not of concern in this matrix
Not of concern in this matrix
(AOAC)
365.1 (NERL)
LRN
Phase Sample Prep
Procedure
TO-13A
Not of concern in this
matrix
TO-13A
TO-13A
TO-13A
TO-6
6002 (NIOSH)
6402 (NIOSH)
Not of concern in this
matrix
Not of concern in this
matrix
LRN
Revision 1.0
A-6
September 29, 2004
-------
Appendix A: Chemical
Analyte(s)
Sarin (GB)
Saxitoxin
Semivolatile Organic
Compounds, NOS
Soman (GD)
Strychnine
Sulfur Dioxide
T-2 Mycotoxins
Tabun (GA)
Tear gas (CS)
chlorobenzylidene malonitrile
Tetanus toxin
Tetraethyl pyrophosphate
CAS No.
107-44-8
35523-89-8
NA
96-64-0
57-24-9
7446-09-5
NA
77-81-6
2698-41-1
NA
107-49-3
Determinative
Technique
GC/MS
HPLC-MS
GC/MS
GC/MS
GC/MS
1C
HPLC-MS
GC/MS
GC/MS
Immunoassay
GC/MS
Determinative Method
Identifier
(SW-846)
994.08 (AOAC)
(SW-846)
8270D (SW-846)
(SW-846)
6004 (NIOSH)
(AOAC)
8270D (SW-846)
8270D (SW-846)
LRN
8270D (SW-846)
Solid Prep
Procedure
3545A/3541
(SW-846)
994.08 (AOAC)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
Not of concern in this matrix
(AOAC)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
LRN
3545A/3541
(SW-846)
Oily Solid
Procedure
3545A/3541/3580A
(SW-846)
994.08 (AOAC)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
Not of concern in this
matrix
994.08 (AOAC)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
LRN
3545A/3541/3580A
(SW-846)
Aqueous and Liquid
Sample Prep Procedure
3520C/3535A
(SW-846)
994.08 (AOAC)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
Not of concern in this matrix
(AOAC)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
LRN
3520C/3535A
(SW-846)
Phase Sample Prep
Procedure
TO-13A
Not of concern in this
matrix
TO-13A
TO-13A
TO-13A
6004 (NIOSH)
Not of concern in this
matrix
TO-13A
TO-13A
LRN
TO-13A
Revision 1.0
A-7
September 29, 2004
-------
Appendix A: Chemical
Analyte(s)
Tetramethylene-
disulfotetramine
Titanium tetrachloride (analyze
for total titanium)
Trimethyl phosphite
VE
VG
VM
Volatile Organic Compounds,
NOS
CAS No.
80-12-6
7550-45-0
121-45-9
21738-25-0
78-53-5
21770-86-5
NA
Determinative
Technique
HPLC-UV
ICP-MS/ICP-AES
GC/MS
GC/MS
GC/MS
GC/MS
GC/MS
Determinative Method
Identifier
8321 B (SW-846)
6020A/6010C
(SW-846)
(SW-846)
8270D (SW-846)
(SW-846)
8270D (SW-846)
(SW-846)
Solid Prep
Procedure
3545A/3541
(SW-846)
3050B (SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
3545A/3541
(SW-846)
(SW-846)
Oily Solid
Procedure
3545A/3541/3580A
(SW-846)
Not of concern in this
matrix
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3545A/3541/3580A
(SW-846)
3585 (SW-846)
Aqueous and Liquid
Sample Prep Procedure
3520C/3535A
(SW-846)
Not of concern in this matrix
3520C/3535A
(SW-846)
Note: For liquid matrices use
3520C (SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
3520C/3535A
(SW-846)
5030C (SW-846)
Phase Sample Prep
Procedure
TO-13A
Not of concern in this
matrix
TO-13A
TO-13A
TO-13A
TO-13A
TO-15
Revision 1.0
September 29, 2004
-------
B:
-------
Page Intentionally Blank
-------
Appendix B: Biological Methods
Agent
Category
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Bacteria
Viruses
Viruses
Viruses
Viruses
Viruses
Viruses
Analyte(s)
Bacillus anthracls
(Anthrax)
Bruceila spp.
(Brucellosis)
Burkhoideria mallei
(glanders)
Burkholdeha
pseudomallei
(melioidosis)
Clostridium perfringens
Francisella tulatensis
(Tularemia)
Coxiella burnetii
(Q-fever)
Salmonella typhl
(Typhoid fever)
Vibrio choleras
(Cholera)
Yersinia pestis (Plague)
Campylobacterjejuni
E. co//O157:H7
Shigeila spp.
Arenavi ruses
Caliciviruses
Nairovirus
Hemorrhagic fever
Rift valley fever
Encephalomyelitis
Waterborne
Determinative
technique
Culture / PCR
Culture
Culture
Culture
Culture
Culture
Culture
Culture/ELISA
Culture
Culture
Culture
Culture
Culture
Biosafety Level 4 -
Ship directly to CDC
laboratory
RT-PCR
Biosafety Level 4 -
Ship directly to CDC
laboratory
Biosafety Level 4 -
Ship directly to CDC
laboratory
Biosafety Level 4 -
Ship directly to CDC
laboratory
Antibody / Integrated
cell culture PCR
Determinative
Method Identifier
LRN
LRN
LRN
LRN
ICR method
LRN
LRN
USDA/FSIS 4
SM 9260H
LRN
SM 9260G
SM 9260F
SM 9260E
Method not identified
AEM Vol. 69 No. 9;
5263-5268
Method not identified
Method not identified
Method not identified
Method not identified
Sample preparation
procedure and/or
sampling method
As specified by LRN
protocol
As specified by LRN
protocol
As specified by LRN
protocol
As specified by LRN
protocol
ICR- As specified in ICR
protocol
As specified by LRN
protocol
As specified by LRN
protocol
As specified by FSIS
protocol
Concentration by placing
Moore swabs in flowing
wastewater for 1 week
As specified by LRN
protocol
Filtration of large volumes
of water using 0.45 or 0.22
micron filter
Collect 100 mL sample in
sterile container
Filtration of large volumes
of water using 0.45 or 0.22
micron filter
As specified by CDC
As specified in AEM Vol.
69 No. 9; 5263-5268
As specified by CDC
As specified by CDC
As specified by CDC
Method not identified
Dustborne
Determinative
technique
Culture / PCR
Culture
Culture
Culture
Culture
Culture
Culture
Culture/ELISA
Culture
Culture
Culture
Culture
Culture
Biosafety Level 4 -
Ship directly to CDC
laboratory
RT-PCR
Biosafety Level 4 -
Ship directly to CDC
laboratory
Biosafety Level 4 -
Ship directly to CDC
laboratory
Biosafety Level 4 -
Ship directly to CDC
laboratory
Antibody/ Integrated
cell culture PCR
Determinative Method
Identifier
OSS3 (OSHA)
LRN
LRN
LRN
ICR method
LRN
LRN
USDA/FSIS 4
SM 9260H
LRN
SM 9260G
SM 9260F
SM 9260E
Method not identified
As specified in AEM
Vol. 69 No. 9; 5263-
5268 (method was
developed for water but
may be adaptable to
other matrices)
Method not identified
Method not identified
Method not identified
Method not identified
Sample preparation
procedure and/or
sampling method
Sampling with wet swabs
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
Swabs, socks, swipes
CDC/NIOSH Sampling
techniques
As specified by CDC
Swabs, socks, swipes
As specified by CDC
As specified by CDC
As specified by CDC
Swabs, socks, swipes
Aerosol
Determinative
technique
Culture / PCR
Culture
Culture
Culture
Culture
Culture
Culture
Culture/ELISA
Culture
Culture
Culture
Culture
Culture
Biosafety Level 4 -
Ship directly to CDC
laboratory
RT-PCR
Biosafety Level 4 -
Ship directly to CDC
laboratory
Biosafety Level 4 -
Ship directly to CDC
laboratory
Biosafety Level 4 -
Ship directly to CDC
laboratory
Antibody / Integrated
cell culture PCR
Determinative
Method Identifier
OSA-7 (OSHA)
LRN
LRN
LRN
ICR method
LRN
LRN
USDA/FSIS 4
SM 9260H
LRN
SM 9260G
SM 9260F
SM 9260E
Method not identified
As specified in AEM
Vol. 69 No. 9; 5263-
5268 (method was
developed for water
but may be
adaptable to other
matrices)
Method not identified
Method not identified
Method not identified
Method not identified
Sample preparation
procedure and/or
sampling method
XMZ/ Anderson Button
Sampler /DFU
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
Wetted-wall cyclones,
CDC/NIOSH Sampling
techniques
As specified by CDC
AGI sampler (modify for
viruses)
As specified by CDC
As specified by CDC
As specified by CDC
AGI sampler (modify for
viruses)
Revision 1.0
September 29, 2004
-------
Appendix B: Biological Methods
Agent
Category
Viruses
Viruses
Viruses
Viruses
Viruses
Protozoa
Protozoa
Protozoa
Analyte(s)
Variola (Smalipox)
Yellow fever
Enterovi ruses
Hepatitis A viruses
VEE virus
Ctyptosporidium
patvum
(Cryptosporidiosis)
Toxoplasma gondii
(Toxoplasmosis)
Entamoeha histolytica
Waterborne
Determinative
technique
Biosafety Level 4 -
Ship directly to CDC
laboratory
Integrated cell culture
PCR
RT-PCR
RT-PCR (HAV)
RT-PCR
FA
RT-PCR
RT-PCR
Determinative
Method Identifier
Method not identified
Method not identified
AEM Vol. 69 No. 6;
3158-3164
AEM Vol. 69 No. 6;
3158-3164
J. Clin. Microbiol. Vol.
38 No. 4;1527-1535
Method 1622, EPA
OW April 2001
Applied and
Environmental
Microbiology, 69
(105:5819-5825
PCR test kits: Mo Bio,
Qiagen
Sample preparation
procedure and/or
sampling method
As specified by CDC
Method not identified
As specified in AEM Vol.
69 No. 6:3158-3164
As specified in AEM Vol.
69 No. 6;3158-3164
As specified in J. Clin.
Microbiol. Vol. 38 No. 4:
1527-1535
Filtration
As specified in Applied anc
Environmental
Microbiology, 69(10)5819
5825
Filtration
Dustborne
Determinative
technique
Biosafety Level 4 -
Ship directly to CDC
laboratory
Integrated cell culture
PCR
RT-PCR
RT-PCR (HAV)
RT-PCR
FA
RT-PCR
RT-PCR
Determinative Method
Identifier
Method not identified
Method not identified
As specified in AEM
Vol. 69 No. 6:3158-
31 64 (method was
developed for water but
may be adaptable to
other matrices)
As specified in AEM
Vol. 69 No. 6:3158-
31 64 (method was
developed for water but
may be adaptable to
other matrices)
Method not identified
Method 1622, EPAOW
April 2001
Applied and
Environmental
Microbiology, 69
(10):5819-5825
PCR test kits: Mo Bio,
Qiagen
Sample preparation
procedure and/or
sampling method
As specified by CDC
Swabs, socks, swipes
Swabs, socks, swipes
Swabs, socks, swipes
Swabs, socks, swipes
Swabs, socks, swipes
Swabs, socks, swipes
Swabs, socks, swipes
Aerosol
Determinative
technique
Biosafety Level 4 -
Ship directly to CDC
laboratory
Integrated cell culture
PCR
RT-PCR
RT-PCR (HAV)
RT-PCR
FA
RT-PCR
RT-PCR
Determinative
Method Identifier
Method not identified
Method not identified
As specified in AEM
Vol. 69 No. 6:3158-
31 64 (method was
developed for water
but may be
adaptable to other
matrices)
As specified in AEM
Vol. 69 No. 6; 31 58-
3164 (method was
developed for water
but may be
adaptable to other
matrices)
Method not identified
Method 1622, EPA
OW April 2001
Applied and
Environmental
Microbiology, 69
(10):5819-5825
PCR test kits: Mo
Bio, Qiagen
Sample preparation
procedure and/or
sampling method
As specified by CDC
AGI sampler (modify for
viruses)
AGI sampler (modify for
viruses)
AGI sampler (modify for
viruses)
AGI sampler (modify for
viruses)
Wetted-wall cyclone
Wetted-wall cyclone
Wetted-wall cyclone
Revision 1.0
September 29, 2004
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