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Rapid sample preparation and bioanalytical techniques for efficient screening of organic pollutants in the environment
Umeå University, Faculty of Science and Technology, Chemistry.
2006 (English)Doctoral thesis, comprehensive summary (Other academic)
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.

Place, publisher, year, edition, pages
Umeå: Kemi , 2006. , 50 p.
Keyword [en]
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
National Category
Environmental Sciences
Identifiers
URN: urn:nbn:se:umu:diva-842ISBN: 91-7264-109-6 (print)OAI: oai:DiVA.org:umu-842DiVA: diva2:144717
Public defence
2006-09-22, KB3B1, KBC, Umeå universitet, Umeå, 10:00 (English)
Opponent
Supervisors
Available from: 2006-09-01 Created: 2006-09-01 Last updated: 2009-12-07Bibliographically approved
List of papers
1. Evaluation of the structure/cross-reactivity relationship of polycyclic aromatic compounds using an enzyme-linked immunosorbent assay kit
Open this publication in new window or tab >>Evaluation of the structure/cross-reactivity relationship of polycyclic aromatic compounds using an enzyme-linked immunosorbent assay kit
2003 (English)In: Analytica Chimica Acta, ISSN 0003-2670, E-ISSN 1873-4324, Vol. 487, no 1, 43-50 p.Article in journal (Refereed) Published
Abstract [en]

A commercially available enzyme-linked immunosorbent assay (ELISA) kit, the PAH soil test, was evaluated with regard to cross-reactivity. Phenanthrene in methanol was used as reference substance. Anthracene, naphthalene and fluorene were chosen as representatives of the 16 US-EPA priority-pollutant polycyclic aromatic hydrocarbons (PAHs). In addition, a number of polycyclic aromatic compounds (PACs), including methyl-, phenyl-, and carbonyl-PAHs, as well as NSO-heterocyclic PACs, found at former industrial sites, were chosen for elucidation of structure/cross-reactivity relationships. The study emphasizes the importance of a priori knowledge of sample composition for accurate interpretation of test results.

Place, publisher, year, edition, pages
Elsevier, 2003
Keyword
Cross-reactivity, PAH, Immunoassay, ELISA, SARs
National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-9232 (URN)10.1016/S0003-2670(03)00466-5 (DOI)
Note

Papers Resulting from the Fifth Workshop on Biosensors and Biological Techniques in Environmental Analysis

Available from: 2008-03-12 Created: 2008-03-12 Last updated: 2017-12-14Bibliographically approved
2. On the semi-quantification of polycyclic aromatic hydrocarbons in contaminated soil by an enzyme-linked immunosorbent assay kit
Open this publication in new window or tab >>On the semi-quantification of polycyclic aromatic hydrocarbons in contaminated soil by an enzyme-linked immunosorbent assay kit
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2006 (English)In: Analytica Chimica Acta, ISSN 0003-2670, E-ISSN 1873-4324, Vol. 555, no 1, 107-113 p.Article in journal (Refereed) Published
Abstract [en]

Polycyclic aromatic hydrocarbons (PAHs) are of environmental concern, for instance when found in contaminated soils at sites where industrial activities have occurred. For efficient screening of such soils, the commercially available enzyme-linked immunosorbent assay (ELISA) kit, the PAH RISc® soil test, can be used. However, the site-specific performance may vary due to differences in soil properties and contamination profiles. Hence, in this study we have examined various contributing factors to the total ELISA measurements uncertainties. These factors include contributions from co-extracted (non-target) compounds, the extraction efficiency and differences in cross-reactivity among the target analytes. Reference values were obtained through pressurized liquid extraction (PLE) and gas chromatography coupled to mass spectrometry (GC–MS) analysis. The results showed that the ELISA does not seem to respond to non-target compounds in the soil extracts to any large extent. Furthermore, high molecular weight PAHs were found to be more efficiently extracted with PLE than with methanol agitation, which is used for ELISA. If this, and the cross-reactivity of the individual PAHs, were taken into consideration, the ELISA and GC–MS results were in good agreement.

Place, publisher, year, edition, pages
Elsevier, 2006
Keyword
Immunoassay; ELISA; Polycyclic aromatic hydrocarbons; Soil; Environmental analysis; Cross-reactivity
National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-12508 (URN)10.1016/j.aca.2005.08.051 (DOI)
Available from: 2007-04-10 Created: 2007-04-10 Last updated: 2017-12-14Bibliographically approved
3. Monitoring dioxins in food and feedstuffs using accelerated solvent extraction with a novel integrated carbon fractionation cell in combination with a CAFLUX bioassay
Open this publication in new window or tab >>Monitoring dioxins in food and feedstuffs using accelerated solvent extraction with a novel integrated carbon fractionation cell in combination with a CAFLUX bioassay
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2005 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 381, no 7, 1472-1475 p.Article in journal (Refereed) Published
Abstract [en]

The concentrations of dioxins in fish oil and fish meal were determined with accelerated solvent extraction, using a novel integrated carbon fractionation extraction cell followed by a miniturized multilayer silica column and bioanalysis on a recently-developed chemically-activated fluorescent gene expression cell bioassay. The developed method allows for simultaneous gravimetric lipid weight determination, which was shown for both matrices under study (about 100% lipid recovery of each sample). Initial results practically meet the quality criteria on screening methods for control of dioxins in food and feedstuffs laid down in the EU Commission Directives 2002/69/EC (food) and 2002/70/EC (feed). This demonstrates that the developed method can be used as a screening tool for monitoring dioxins in food and feed after some additional improvements and testing on a greater number of matrices.

Keyword
Dioxin, Accelerated solvent extraction, ASE, Cell-based bioassay, CAFLUX
National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-13066 (URN)10.1007/s00216-005-3117-4 (DOI)15821857 (PubMedID)
Available from: 2007-05-02 Created: 2007-05-02 Last updated: 2017-12-14Bibliographically approved
4. Rapid screening of dioxin-contaminated soil by accelerated solvent extraction/purification followed by immunochemical detection
Open this publication in new window or tab >>Rapid screening of dioxin-contaminated soil by accelerated solvent extraction/purification followed by immunochemical detection
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2006 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 385, no 2, 357-366 p.Article in journal (Refereed) 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.

Place, publisher, year, edition, pages
Springer Science+Business Media B.V., 2006
Keyword
Immunoassay, ELISA, Soil, Dioxin, Accelerated solvent extraction
National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-11789 (URN)10.1007/s00216-006-0402-9 (DOI)16598456 (PubMedID)
Available from: 2007-08-17 Created: 2007-08-17 Last updated: 2017-12-14Bibliographically approved
5. Analysis of dioxins in contaminated soils using the CALUX and CAFLUX bioassays, an immunoassay and gas chromatography/high-resolution mass spectrometry
Open this publication in new window or tab >>Analysis of dioxins in contaminated soils using the CALUX and CAFLUX bioassays, an immunoassay and gas chromatography/high-resolution mass spectrometry
Show others...
Manuscript (Other academic)
Identifiers
urn:nbn:se:umu:diva-5261 (URN)
Available from: 2006-09-01 Created: 2006-09-01 Last updated: 2010-01-13Bibliographically approved

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