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Rapid screening of dioxin-contaminated soil by accelerated solvent extraction/purification followed by immunochemical detection
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
Department of Entomology and Cancer Research Center, University of California, USA.
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
Vise andre og tillknytning
2006 (engelsk)Inngår i: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 385, nr 2, s. 357-366Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Since soils at industrial sites might be heavily contaminated with polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), there is a need for large-scale soil pollution surveys and, thus, for cost-efficient, high-throughput dioxin analyses. However, trace analysis of dioxins in complex matrices requires exhaustive extraction, extensive cleanup, and very sensitive detection methods. Traditionally, this has involved the use of Soxhlet extraction and multistep column cleanup, followed by gas chromatography-high-resolution mass spectrometry (GC/HRMS), but bioanalytical techniques may allow much more rapid, cost-effective screening. The study presented here explores the possibility of replacing the conventional method with a novel approach based on simultaneous accelerated solvent extraction (ASE) and purification, followed by an enzyme-linked immunosorbent assay (ELISA). Both the traditional and the novel cleanup and detection approaches were applied to contaminated soil samples, and the results were compared. ELISA and GC/HRMS results for Soxhlet-extracted samples were linearly correlated, although the ELISA method slightly underestimated the dioxin levels. To avoid an unacceptable rate of false-negative results, the use of a safety factor is recommended. It was also noted that the relative abundance of the PCDDs/PCDFs, evaluated by principal component analysis, had an impact on the ELISA performance. To minimize this effect, the results may be corrected for differences between the ELISA cross-reactivities and the corresponding toxic equivalency factor values. Finally, the GC/HRMS and ELISA results obtained following the two sample preparation methods agreed well; and the ELISA and GC/HRMS results for ASE extracts were strongly correlated (correlation coefficient, 0.90). Hence, the ASE procedure combined with ELISA analysis appears to be an efficient approach for high-throughput screening of PCDD-/PCDF-contaminated soil samples.

sted, utgiver, år, opplag, sider
Springer Science+Business Media B.V., 2006. Vol. 385, nr 2, s. 357-366
Emneord [en]
Immunoassay, ELISA, Soil, Dioxin, Accelerated solvent extraction
HSV kategori
Identifikatorer
URN: urn:nbn:se:umu:diva-11789DOI: 10.1007/s00216-006-0402-9PubMedID: 16598456OAI: oai:DiVA.org:umu-11789DiVA, id: diva2:151460
Tilgjengelig fra: 2007-08-17 Laget: 2007-08-17 Sist oppdatert: 2019-04-09bibliografisk kontrollert
Inngår i avhandling
1. PLE with integrated clean up followed by alternative detection steps for cost-effective analysis of dixons and dioxin-like compounds
Åpne denne publikasjonen i ny fane eller vindu >>PLE with integrated clean up followed by alternative detection steps for cost-effective analysis of dixons and dioxin-like compounds
2008 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) are two structurally related groups of chemicals, generally referred to as `dioxins´. These are of great concern due to their high toxicity and global spread. Other groups of compounds with similar chemical structure and toxicity mechanisms are the brominated analogues polybrominated dibenzo-p-dioxins (PBDDs) and polybrominated dibenzofurans (PBDFs), and the dioxin-like polychlorinated biphenyls (PCBs). Numerous studies have been undertaken to investigate sources and transport routes of dioxins. However, much remains to be done, including analytical, inventories of dioxin-like compounds, such as PBDD/Fs, and the development of more convenient analytical methods. The currently standard procedure for analyzing dioxins (and dioxin-like compounds) is to use Soxhlet extraction followed by multi-step clean-up and gas chromatography - high resolution mass spectrometry (GC- HRMS) for detection. Unfortunately, this method is very solvent, labor and time-consuming, making it very expensive.

The main aim of the studies this thesis was to develop pressurized liquid extraction (PLE) with integrated clean up techniques for fast, convenient preparation of dioxin samples. PLE with integrated clean-up has previously been used for extracting dioxins from biological samples, but in these studies the possibility of extending its use to abiotic samples was explored. The results show that PLE with an integrated carbon trap is suitable for analyzing dioxins in various types of soil samples, sediment and flue gas samples. The results also showed that it has potential for analyzing dioxins in fly ash. The thesis focuses on developments of the methodology for dioxin analysis, but also includes results obtained from PBDDs and dioxin-like PCB analyses. In addition, the possibility of using various other kinds of detection techniques rather than GC-HRMS, such as enzyme-linked immunosorbent assays (ELISAs) or two-dimensional gas chromatography with micro electron capture detection (GCxGC-µECD) was explored. The results indicate that ELISA and GCxGC-µECD could serve as complementary detection systems in some cases. However, it is not yet possible to fully replace GC-HRMS.

A further refinement of the PLE with in-cell clean-up technique is the modular approach developed in these studies. With this technique it is possible to include various steps for both clean-up and fractionation. For example, sulphuric acid impregnated silica could be combined with active carbon for the simultaneous removal of lipids (along with other interferences) and fractionation of PCBs and PCDD/Fs. It was shown that the method could provide data that agreed reasonably well with both reference values and values obtained using traditional methods. In general PLE proved to have high extraction efficiency and to yield very similar congener profiles to the reference method. In addition, it was shown that it allowed one-step extraction and clean-up of a salmon sample. Such single-step procedures are the ultimate goals for any extraction technique, and it would be highly desirable to develop one-step methods that could be extended to other types of samples. For the rest of the matrices tested (soil, sediment, mussel and crab tissue and flue gas) the method was successful, however a final polishing step is currently required, involving either dilution or clean-up using miniaturized multilayer silica columns, to obtain extracts that are pure enough for GC-HRMS analysis.

Using the developed modular-PLE system substantial costs could be saved. It was estimated that the method could reduce the cost of preparing samples by up to 90%, which would greatly facilitate large-scale inventories.

sted, utgiver, år, opplag, sider
Umeå: Kemi, 2008. s. 37
Emneord
Pressurized liquid extraction (PLE), PLE-C, modular PLE, ELISA, dioxin, dioxin-like compounds, GCxGC, analytical methods
HSV kategori
Identifikatorer
urn:nbn:se:umu:diva-1665 (URN)978-91-7264-584-4 (ISBN)
Disputas
2008-06-13, N360, Naturvetarhuset, Umeå, 13:00 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2008-05-19 Laget: 2008-05-19 Sist oppdatert: 2009-07-02bibliografisk kontrollert
2. Rapid sample preparation and bioanalytical techniques for efficient screening of organic pollutants in the environment
Åpne denne publikasjonen i ny fane eller vindu >>Rapid sample preparation and bioanalytical techniques for efficient screening of organic pollutants in the environment
2006 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Large numbers of samples often need to be prepared and analysed in surveys of organic pollutants in the environment, but while the methods commonly used in such surveys can provide abundant detail they are generally costly, time-consuming and require large amounts of resources, so there is a need for simpler techniques. The work underlying this thesis assessed the potential utility of more convenient sample preparation and bioanalytical techniques for rapidly screening various environmental matrices that could be useful complements to higher resolution methods.

Initially, the utility of a simplified extraction technique followed by an enzyme-linked immunosorbent assay (ELISA) for detecting polycyclic aromatic hydrocarbons (PAHs) in authentic (i.e. unspiked) contaminated soils was explored. The results showed that there are relationships between the structure and cross-reactivity among compounds that often co-occur with target PAHs. However, their potential contribution to deviations between estimates of total PAH contents of soils obtained using ELISA and gas chromatography-mass spectrometry (GC-MS) based reference methods were limited. Instead, the cross-reactivity of target PAHs and the failure to extract all of the PAHs prior to the ELISA determinations were the main reasons for these deviations.

Polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were detected in food and feed matrices, as well as in authentic contaminated soils using different bioanalytical techniques – ELISA and two cell-based bioassays: CAFLUX and CALUX (chemically activated fluorescent/luciferase gene expression) assays. In addition, enhanced sample preparation techniques based on accelerated solvent extraction (ASE) were developed. ASE with integrated carbon fractionation (ASE-C) in combination with CAFLUX produced estimates of PCDD and PCDF contents in fish oil and fish meal that agreed well with results obtained using reference methods. Furthermore, results from ELISA and GC-high resolution MS analyses of extracts of PCDD- and PCDF-contaminated soil samples obtained using an adjusted ASE-C technique were strongly correlated.

Finally, the thesis reports the first experiments in which the results of CAFLUX, CALUX, and ELISA determinations of PCDDs and PCDFs in extracts of authentic contaminated soil samples were evaluated and compared to those obtained using a reference method. All of the bioanalytical techniques were found to be sufficiently sensitive, selective, and accurate for use in screening in compliance with soil quality assessment criteria. Overall, the improved sample preparation and bioanalytical techniques examined proved to be useful potential complements to conventional methods, enhancing the analytical framework for PAHs, PCDDs, and PCDFs. However, further validation has to be undertaken before they are applied on a large-scale.

sted, utgiver, år, opplag, sider
Umeå: Kemi, 2006. s. 50
Emneord
accelerated solvent extraction, CAFLUX, food, immunoassay, PAH, PCDD, PCDF, pressurized liquid extraction, soil, CALUX, cell-based bioassay, chemically activated fluorescent gene expression, chemically activated luciferase gene expression, dioxin, ELISA, enzyme-linked immunosorbent assay, feed
HSV kategori
Identifikatorer
urn:nbn:se:umu:diva-842 (URN)91-7264-109-6 (ISBN)
Disputas
2006-09-22, KB3B1, KBC, Umeå universitet, Umeå, 10:00 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2006-09-01 Laget: 2006-09-01 Sist oppdatert: 2009-12-07bibliografisk kontrollert

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