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Detection and Site Characterization

Determining the presence of dioxins at a site can be a matter of simple screening. However, determining which dioxins are present involves speciation testing. This section lists well-established, standard methods commonly used for detecting, measuring, and/or monitoring dioxins particularly EPA-approved methods used for environmental samples.

Check the National Environmental Methods Index (NEMI), for a free, searchable clearinghouse of methods and procedures for regulatory and non-regulatory analyses.

Analytical Methods

Adobe PDF LogoDioxin and Furan Analytical Services (DLM02.2)
U.S. EPA, (PDF)(4 pp, 421 K)

This fact sheet provides information on the analysis of seventeen 2,3,7,8-substituted tetra- through octa- chlorinated dibenzo-dioxins and chlorinated dibenzo-furans in water, soil, sediment, sludge, non-human tissue, ash, oil, and oily matrices. Since this service is not one of the analyses conducted under the Superfund Contract Laboratory Program (CLP), it is managed as part of the Non-Routine Analytical Services program.

Adobe PDF LogoPolychorinated Biphenyl Congeners Analytical Services (CBC01.2)
U.S. EPA, (PDF)(3 pp, 382 K)

This fact sheet provides information on the analysis of 209 substituted mono- through deca- polychlorinated biphenyl (PCB) congeners, which include dioxin-like PCB congeners, in water, soil, sediment, sludge, non-human tissue, ash, oil, and oily matrices.

Dioxins, Furans, PCBs, and Congeners Analytical Service within the Superfund Contract Laboratory Program

This high-resolution service provides for the detection, and quantitative measurement of CBCs (either the 12 congeners defined as toxic by the World Health Organization or all 209 congeners) and/or 17 2,3,7,8-substituted tetra- through octachlorinated CDDs and CDFs in aqueous/water, soil/sediment, sludge, tissue (non- human), biosolids, ash, oil, and oily matrices using high resolution gas chromatography/high resolution mass spectrometry to analyze samples. This service offers preliminary results and 21- and 35-day turnaround.

Soil, Sediment, Water, and Other Environmental Matrices

Test Methods for Evaluating Solid Wastes: Physical/Chemical Methods, 3rd Edition
U.S. EPA, SW-846.

Method 4025: Screening for Polychlorinated Dibenzodioxins and Polychlorinated Dibenzofurans (PCDD/Fs) by Immunoassay

Method 4025 documents the procedure for the analysis of polychlorinated dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in soil at 500 parts per trillion (picograms per gram) using a simple one-step liquid phase oxidative cleanup or, at mid to low ppt levels, using a two-step coupled column cleanup (oxidation and activated carbon binding) adapted from Method 8290.

Method 4425: Screening Extracts of Environmental Samples for Planar Organic Compounds (PAHs, PCBs, PCDDs/PCDFs) by a Reporter Gene on a Human Cell Line [draft update IVB]

Method 4425 provides information on the screening procedure that will detect the total amount of planar compounds – including PCDDs, PCDFs, coplanar PCBs, and high molecular weight PAHs – in solvent extracts of environmental samples of soil, sediment, tissue, and water.

Method 8280B: Polychlorinated Dibenzo-p-Dioxins and Polychlorinated Dibenzofurans by High Resolution Gas Chromatography/Low Resolution Mass Spectrometry (HRGC/LRMS)

This document outlines the method used in the detection and measurement of PCDDs and PCDFs in water, soil, fly ash, and chemical waste samples, including stillbottom, fuel oil, and sludge matrices.

Method 8290A: Polychlorinated Dibenzodioxins (PCDDs) and Polychlorinated Dibenzofurans (PCDFs) by High-Resolution Gas Chromatography/High-Resolution Mass Spectrometry (HRGC/HRMS)

Method 8290A provides procedures for the detection and quantitative measurement of polychlorinated dibenzo-p-dioxins (tetra- through octachlorinated homologues; PCDDs), and polychlorinated dibenzofurans (tetra- through octachlorinated homologues; PCDFs) in a variety of environmental matrices and at part-per-trillion (ppt) to part-per-quadrillion (ppq) concentrations.

Air Emissions

Method 23 Determination of Polychlorinated Dibenzo-p-Dioxins and Polychlorinated Dibenzofurans from Stationary Sources
U.S. EPA, Air Emission Measurement Center, 2017

SW-846 Method 0023A: Sampling Method for Polychlorinated Dibenzo-p-Dioxin and Polychlorinated Dibenzofuran Emission from Stationary Sources

This method describes the sampling procedure for determining stack emissions of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) from stationary sources.

Adobe PDF LogoMethod 613 Methods for Organic Chemical Analysis of Municipal -2,3,7,8-Tetrachlorodibenzo-p-Dioxin
U.S. EPA, 40 CFR 136, Appendix A: Methods for Organic Chemical Analysis of Municipal and Industrial Wastewater.

Adobe PDF LogoCompendium of Methods for the Determination of Toxic Organic Compounds in Ambient Air Second Edition Compendium Method TO-9A Determination Of Polychlorinated, Polybrominated and Brominated/Chlorinated Dibenzo-p-Dioxins And Dibenzofurans In Ambient Air
U.S. EPA, Office of Research and Development, EPA/625/R-96/010b, 94 pp. January 1999


Adobe PDF LogoMethod 1613: Tetra- Through Octa- Chlorinated Dioxins and Furans by Isotope Dilution HRGC/HRMS
U.S. EPA, Office of Water. EPA 821-B-94-005, 89 pp, Revision B, 1994.

Method 1614 was developed by the U.S. EPA for isomer-specific determination of the 2,3,7,8-substituted tetra- through octa-chlorinated dibenzo-p-dioxins and dibenzofurans in aqueous, solid, and tissue matrices by isotope dilution, high resolution capillary gas chromatography/high resolution mass spectrometry.


Ion-Trap Tandem Mass Spectrometry for the Analysis of Polychlorinated Dibenzo-p-dioxins, Dibenzofurans, and Dioxin-like Polychlorinated Biphenyls in Food
Journal of Agricultural and Food Chemistry 55(26):10531-10539(2007)
Jessica Malavia, et al.

An improved clean-up strategy for simultaneous analysis of polychlorinated dibenzo-p-dioxins (PCDD), polychlorinated dibenzofurans (PCDF), and polychlorinated biphenyls (PCB) in fatty food samples
Catherine Pirard, Jean-François Focant, and Edwin De Pauw
Anal Bioanal Chem (2002) 372 :373-381

Dioxins in Food and Feed – Reference Methods and New Certified Reference Materials (DIFFERENCE)
RIVO-Netherlands Institute for Fisheries Research. 75pp, 2002


Toward Streamlined Identification of Dioxin-like Compounds in Environmental Samples through Integration of Suspension Bioassay
Environmental Science & Technology 51(6):3382-3390(2017)
DOI: 10.1021/acs.est.6b06003
Hongxia Xiao, Markus Brinkmann, Beat Thalmann, Andreas Schiwy, Sigrid Grosse Brinkhaus, et al.

A high-throughput effect-directed analysis (HT-EDA) work-flow combining reversed phase high-performance liquid chromatography fractionation of samples into 96-well microplates was proposed, followed by toxicity assessment in the micro-EROD bioassay with the wild-type rat hepatoma H4IIE cells, and chemical analysis of bioactive fractions.

Adobe PDF LogoSoil Dioxin Relative Bioavailability Assay Evaluation Framework
U.S. Environmental Protection Agency, OSWER 9200.2-136, 16 pp, 2015

Until standard procedures for estimating the relative bioavailability (RBA) of PCDD/F in soil are established, a consistent approach is needed to evaluate the strengths and weaknesses of assays designs proposed or implemented to support risk assessments. This report offers a framework for making such evaluations.

Final Report on the Performance of the Eichrom Technologies Procept® Rapid Dioxin Assay for Soil and Sediment Samples
EPA 540-R-11-002, 48 pp, 2011
Stephen Billets

This report describes the experimental design of the follow-on site-specific study. Samples from additional sites were included in the study, and Eichrom utilized an optimized method for its Procept® Rapid Dioxin Assay analyses.

Enzyme-Linked Immunosorbent Assay for Screening Dioxin Soil Contamination by Uncontrolled Combustion During Informal Recycling in Slums
Environ Toxicol Chem. Nov; 27(11): 2224-2232(2008).
Trindade, M., M. Nording, M. Nichkova, E. Spinnel, P. Haglund, M.S. Last, S. Gee, B. Hammock, J.A. Last, G. Gonzalez-Sapienza, and B.M. Brena.

The present study describes a first approach toward the use of a dioxin antibody-based enzyme-linked immunosorbent assay (ELISA) as the basis for a monitoring program to assess the toxicological impact of uncontrolled combustion in slums.

An Enzyme-Linked Immunosorbent Assay for the Determination of Dioxins in Contaminated Sediment and Soil Samples
Chemosphere 72(1):95-103(2008)
J.M. Van Emon, J.C. Chuang, R.A. Lordo, M.E. Schrock, M. Nichkova, S.J. Gee, B.D. Hammock.

A 96-microwell enzyme-linked immunosorbent assay (ELISA) method was evaluated to determine PCDDs/PCDFs in sediment and soil samples from an EPA Superfund site.

Analysis of Dioxins in Contaminated Soils with the Calux and Caflux Bioassays, an Immunoassay, and Gas Chromatography/High-Resolution Mass Spectrometry
Environmental Toxicology and Chemistry 26(6): 1122-1129(2007)
M. Nording, M.S. Denison, D. Baston, Y. Persson, E. Spinnel, P. Haglund.

The performance of three bioanalytical methods (chemically activated luciferase expression assay, chemically activated fluorescence expression assay, and the enzyme-linked immunosorbent assay (ELISA)) were assessed for quantifying PCDD/Fs in aliquots of extracts from aged-contaminated soil samples. The results were compared those obtained by GC/HRMS.

Dioxin and Dioxin-Like Compounds in Soil and Sediment
Billets, S.N. and A.B. Dindal.
EPA 600-R-04-036, 2004.

In 2004 a demonstration of technologies for determining the presence of dioxin in soil and sediment was conducted under the U.S. Environmental Protection Agency Superfund Innovative Technology Evaluation Program in Saginaw, Michigan, at Green Point Environmental Learning Center. The primary purpose of the demonstration was to evaluate innovative monitoring technologies:

  • AhRC PCR™ Kit, Hybrizyme Corporation
  • Ah-IMMUNOASSY® Kit, Paracelsian, Inc.
  • Coplanar PCB Immunoassay Kit, Abraxis LLC
  • DF-l Dioxin/Furan Immunoassay Kit, CAPE Technologies LLC
  • CALUX® by Xenobiotic Detection Systems, Inc.
  • Dioxin ELISA Kit, Wako Pure Chemical Industries LTD

Adobe PDF LogoAnalysis of Uncertainty in Estimating Dioxin Bioaccumulation Potential in Sediment-Exposed Benthos
U.S. Army Corps of Engineers, ERDC TN-DOER-R5, 18 pp. September 2004

Adobe PDF LogoBiomarker-Based Analysis for Contaminants in Sediments/Soil: Review of Cell-Based Assays and cDNA Arrays
U.S. Army Corps of Engineers, Dredging Operations Environmental Research Program. ERDC TN-DOER-C19, 11 pp, 2000.

Adobe PDF LogoProtocols for a Rapid Clean-Up/Extraction Procedure and an Improved P450RGS Dioxin Screening Assay for Sediments
U.S. Army Corps of Engineers, Dredging Operations Environmental Research Program. ERDC TN-DOER-C10, 28 pp, 2000.

Adobe PDF LogoComparison of Two Cell-Based Assays for Screening Dioxin and Dioxin-Like Compounds in Sediments
U.S. Army Corps of Engineers, Dredging Operations Environmental Research Program. Technical Note DOER-C8, 12 pp, 1999.

Importance of Clean-up for Comparison of TEQ-values Obtained by CALUX and Chemo-analysis
V. Wouwea, et al.
Talanta 63 (2004) 1269-1272, 4pp

Sampling and Site Characterization

Site Evaluation for Dioxin at Superfund Sites

EPA website lists guidance documents on the sampling for dioxin at Superfund site and analyses under the Contract Laboratory Program.

Adobe PDF LogoField Testing a Passive Multisampler to Measure Dioxins/Furans and Other Contaminant Bioavailability in Aquatic Sediments - Phase II
Lohmann, R., and M. Weinstein. SERDP Project ER-2538, 58 pp, 2022

An in situ passive multi-sampler was further developed and field tested to quantify polychlorinated dibenzo-p-dioxins and dibenzofurans and PCBs in sediment and water column. The novel sampler was validated and field-tested in shallow and deep sediments at several sites along the Passaic River and Newark Bay. Our The technical approach consisted of the construction, deployment, and validation of a modified sediment porewater multisampler for dioxins/furans and other HOCs. The construction of a passive multisampler in the flukes of a Danforth-style anchor was optimized in field tests. Initial field trials needed a diver to ensure deployments, while the final field deployment proceeded without divers. Performance reference compounds were included to correct for the lack of equilibrium of target hydrophobic organic contaminants (HOCs) during field deployments. Sediment grabs were collected at each site to derive HOC porewater concentrations through ex situ equilibrations. The deployment trial in Newark Bay included a comparison to a previously circular sampler and sediment equilibrations for PCBs and PCDD/Fs. The retrieval rate of the Danforth anchors exceeded 80%, assisted by using extra anchor chain. At all sites, porewater concentrations derived from ex situ equilibrations exceeded in situ passive samplers. Comparing ex situ versus in situ porewater concentrations in a second field trial showed ex situ concentrations of PCBs exceeded in situ concentrations by two to three-fold. At most sites, PCDD/Fs were below detection limits. A newly published standard protocol was followed for the ex situ determination of HOC porewater concentrations that prevented depletion of porewater, which may have contributed to the observed discrepancy. A reanalysis of foodweb samples from the Passaic River suggests that passive sampling may be a powerful tool to predict HOC concentrations in the storage lipids of biota.

Using Polyethylene Passive Samplers to Study the Partitioning, Fluxes and Bioaccumulation of PBDEs in an Urban River
Environmental Science & Technology, 51(16):9062-9071(2017)
M. Khairy and R. Lohmann.

To better understand the biogeochemical controls on PBDEs, 12 PBDE congeners were measured in air, water, sediment and porewater of the lower Passaic River.

Adobe PDF LogoIn Situ Laser-Induced Fluorescence: Novel Applications for Contaminated Sediments
RemTech 2019: Remediation Technologies Symposium, 16-18 October, Banff, 18 slides, 2019
Sweet, B., K. Davidson, and C. Lake.

The potential of an in situ laser-induced fluorescence technique to spatially delineate organic-rich industrial sediments contaminated with dioxins and furans to collect unique optical signatures of materials based on their physiochemical properties was explored. Lab and field work were conducted at a stabilization basin that has received industrial wastewater over the past 50+ years in Nova Scotia, Canada.

Comprehensive Analysis of the Great Lakes Top Predator Fish for Novel Halogenated Organic Contaminants by GCxGC-HR-ToF Mass Spectrometry
Environmental Science & Technology 52(5):2909-2917(2018) DOI: 10.1021/acs.est.7b05999
S. Fernando, A. Renaguli, M.S. Milligan, J.J. Pagano, P.K. Hopke, T.M. Holsen, and B.S. Crimmins.

Acetylcholinesterase Is a Potential Biomarker for a Broad Spectrum of Organic Environmental Pollutants
Environmental Science & Technology, 52, 15, 8065-8074(2018) (Critical Review) DOI:10.1021/acs.est.7b04004
H. Fu, Y. Xia, Y. Chen, T. Xu, L. Xu, Z. Guo, H. Xu, H.Q. Xie, and B. Zhao

This review summarizes correlations of Acetylcholinesterase (AChE, EC activity of certain organisms with the level of the contaminants in particular habitats, disruptions of AChE activity upon treatment with the emerging disruptors in vivo and in vitro, and action mechanisms underlying the effects on AChE. Over 40 chemicals belonging to six main categories were reviewed, including 12 POPs listed in the Stockholm Convention.

Adobe PDF LogoDevelopment of a Passive Multisampling Method to Measure Dioxins/Furans and Other Contaminant Bioavailability in Aquatic Sediments
R. Lohmann
SERDP Project ER-2538, 72 pp, 2016

A passive sampling method utilizing polyethylene samplers was developed for in situ sampling of dioxins/furans and a wide range of other hydrophobic organic contaminants in sediment and the overlying water column.

Tracking the Spatial Fate of PCDD/F Emissions from a Cement Plant by Using Lichens as Environmental Biomonitors
Environmental Science & Technology 50(5):2434-2441(2016)
DOI: 10.1021/acs.est.5b04873
S. Augusto, P. Pinho, A. Santos, M.J. Botelho, J. Palma-Oliveira, and C. Branquinho

The use of lichens to track minor sources of air pollution was tested by transplanting lichens from a background area located in a naural park in the vicinity of a cement manufacturing plant that uses alternative fuel and in an area surrounded by other important sources of pollution. After 7 months of exposure, the lichens were collected and analyzed for 17 PCDD/F congeners.

Adobe PDF LogoStandardized Toolkit for Identification and Quantification of Dioxin and Furan Releases
United Nations Environment Programme, 250 pp, 2nd ed. February 2002

Adobe PDF LogoNational Functional Guidelines for Chlorinated Dibenzo-p-Dioxins (CDDs) and Chlorinated Dibenzofurans (CDFs) Data Review
U.S. EPA Analytical Services Branch
EPA 540-R-11-016, OSWER 9240.1-53, 62 pp, 2011.

Dioxin at Superfund Sites
U.S. EPA Office of Superfund Remediation and Technology Innovation

Advanced Monitoring Systems Center Environmental Technology Verification (ETV) Program (Archived)

Technologies for Monitoring and Measurement of Dioxin and Dioxin-Like Compounds in Soil and Sediment: Superfund Innovative Technology Evaluation (SITE) Program Innovative Technology Verification Reports