戴奧辛類化合物為環境中具有嚴重危害之化合物之一，傳統高解析氣相層析質譜儀（HRGC/HRMS）之分析費用昂貴耗時，本研究之主要目的為發展一快速便宜之生物檢測方法。本研究採用之戴奧辛類化合物生物檢測法（以下簡稱生物檢測法），為化學活性冷光酵素基因表現法(chemical activated luciferase gene expression)，簡稱為CALUX生物檢測法。
本研究之內容主要分為三部分：一、將現有文獻資料之生物檢測法，做有組織之整理比較，顯示國際上生物檢測法相關研究之優點及缺失，並提出未來可行之重要研究方向；二、建立牛奶中戴奧辛類化合物之生物檢測法及相關之品質保證及品質控制（QA/QC）；三、探討環境中常見之八種重金屬離子（Hg2+、Ag+、Cu2+、Fe2+、As5+、Cr6+、Cd2+和Pb2+）對生物檢測法之影響。
第一部分之研究結果顯示：主要之戴奧辛生物檢測法是以抗原與抗體免疫分析、生物體對戴奧辛新陳代謝反應(含基因重組細胞株)、結合電子儀器之生物感測器等種類。而其中部分免疫分析與新陳代謝反應均已被美國環保署公告為標準戴奧辛分析方法之一，足可見其具穩定性及準確性均已達到可應用於實際樣品之要求。生物感測器雖然尚未被列入美國環保署公告方法之中，但其可於現場即時監測之特性，亦為未來生物檢測法發展重點之一。
第二部分之研究結果顯示：可設定CALUX生物檢測法之品管標準為：(1)品質管制標準(QC standard)為算術平均值±2倍標準差(μ ± 2σ)之範圍；(2)回收率之標準偏差(relative standard deviation, RSD)於20.7%內。以28個市面上銷售之牛乳樣品進行量測，其結果顯示CALUX生物檢測法具有良好之穩定性，足以作為篩選大量牛乳樣品之可靠工具。
第三部分之研究結果顯示：特定的金屬為CALUX潛在之干擾因子，其細胞株(cell-line)對特定的金屬具有高度之敏感性。因子實驗結果顯示在常見之8種重金屬離子中(Hg2+、Ag+、Cu2+、Fe2+、As5+、Cr6+、Cd2+和Pb2+)，Hg2+、Cu2+與Cr6+對H1L6.1細胞株之冷光活性表現具有統計上之顯著差異。此外，冷光酵素活性與H1L6.1細胞之存活率並無明確之相關性存在，表示除了細胞受到重金屬毒害為可能影響因子外，尚有其他影響CYP1A活性表現之機制存在。
由本研究之實驗結果顯示，戴奧辛類化合物以生物檢測法分析，可達到快速及便宜之目的，唯環境中之干擾因子如重金屬之影響必須先行釐清，方能於前處理步驟中以相應之處理程序去除干擾物。以生物檢測法來量測戴奧辛類化合物為一甚為可行之量測方式。以量測戴奧辛類化合物代謝產物量之生物檢測方法，適合搭配藥物動力學等相關模式進行進階之風險評估；而生物檢測器配合可隨身攜帶之量測儀器，則可成為現場快速監測之良好工具。

The dioxin-like compounds are one of the most hazardous material groups in the world. Traditional chemical analysis methods (e.g., HRGC/HRMS and HRGC/LRMS) are time-consuming and costly for these compounds. Thus, the major aim of this study is to build a rapid and less-cost bioassay method. The Xenobiotic Detection Systems - chemical activated luciferase gene expression (CALUX) bioassay was thus employed in this study for detecting dioxin-like compunds.
The dissertation is mainly concerned with three issues: 1) a review of the bioassay literature; 2) building the bioassay method for detecting dioxin-like compounds in milk and relative quality assurance/ quality control (QA/QC); and 3) discussing the effects of eight metal ions (Hg2+, Ag+, Cu2+, Fe2+, As5+, Cr6+, Cd2+, and Pb2+) on the bioassay method.
The results of part one show that the main dioxin bioassay methods are immunoassay, the biological metabolic reaction of dioxins (including the DNA recombinant cell), and biosensors. The methods that based on the first and second principles have been accepted as standard analysis methods by U.S. EPA, and thus they are stable and accurate enough for use with real samples. Although biosensors are not a standard analysis method used by the U.S. EPA, however, they are important because they offer real-time on-site monitoring.
The results of part two show that the analyzed data suggest that the CALUX bioassay criteria have: (1) control chart for quality control (QC) standards within the μ ± 2σ range; and (2) a recovery efficiency (i.e., relative standard deviation (RSD) of 20.7%. This bioassay method was utilized on 28 commercially available pasteurized milk samples, and the results illustrate that CALUX is a powerful approach for screening a large number of such samples.
Finally, the results of part three show further study found that the bioassay cell-line was sensitive to particular metals, which might be a potential interfering factor. A two-level factorial-designed experiment was undertaken to identify the statistically significant factors from eight metal ions (Hg2+, Ag+, Cu2+, Fe2+, As5+, Cr6+, Cd2+, and Pb2+), and the results revealed that Hg2+, Cu2+, and Cr6+ had significant effects on the luciferase activity expression of the H1L6.1 cells. In addition, the correlation between luciferase activity and the relative survival ratio of H1L6.1 cells with metal concentrations was not comparable. This reveals the inhibiting effect of metals on luciferase activity may not only be caused by the reduction of H1L6.1 cells, but may also be caused by the other mechanisms, such as a reduction of luciferase enzyme production or the inhibiting effect of metal ions on CYP1A activation.
The study illustrates that the bioassay method it presents is a rapid and less-costly approach for detecting dioxin-like compounds. However, potential interference factors, (specifically heavy metal ions) for the bioassay should be eliminated in the pretreatment process to maintain the stability of analysis. The proposed bioassay method with metabolite assay could be emplyeed in risk assessment by using the physiologically based pharmacokinetic model; in addition, the biosensors could be used as an on-site monitoring tool when combined with portable instruments.

Alcock, R.E.; Sweetman, A.J.; Anderson, D.R.; Fisher, R.; Jennings, R.A.; Jones, K.C., (2002) Using PCDD/F congener patterns to determine the source of elevated TEQ concentrations in cows milk: a case study. Chemosphere, Vol.46, No.3, pp.383-391.
Asari, M.; Takatsuki, H.; Yamazaki, M.; Azuma, T.; Takigami, H.; Sakai, S.I., (2004) Waste wood recycling as animal bedding and development of bio-monitoring tool using the CALUX assay. Environment International, Vol.30, No.5, pp.639-649.
Ayotte, P.; Dewailly, E.; Lambert, G.H.; Perkins, S.L.; Poon, R.; Feeley, M.; Larochelle, C.; Pereg, D., (2005) Biomarker measurements in a coastal fish-eating population environmentally exposed to organochlorines. Environmental Health Perspectives, Vol.113, No.10, pp.1318-1324.
Behnisch, P.A.; Hosoe, K.; Sakai, S.-i., (2001a) Bioanalytical screening methods for dioxins and dioxin-like compounds -- a review of bioassay/biomarker technology. Environment International, Vol.27, No.5, pp.413-439.
Behnisch, P.A.; Hosoe, K.; Sakai, S., (2001b) Combinatorial bio/chemical analysis of dioxin and dioxin-like compounds in waste recycling, feed/food, humans/wildlife and the environment. Environment International, Vol.27, No.6, pp.495-519.
Behnisch, P.A.; Hosoe, K.; Brouwer, A.; Sakai, S., (2002a) Screening of dioxin-like toxicity equivalents for various matrices with wildtype and recombinant rat hepatoma H4IIE cells. Toxicological Sciences, Vol.69, No.1, pp.125-130.
Behnisch, P.A.; Hosoe, K.; Shiozaki, K.; Kiryu, T.; Komatsu, K.; Schramm, K.W.; Sakai, S., (2002b) Melting and incineration plants of municipal waste - Chemical and biochemical diagnosis of thermal processing samples (emission, residues). Environmental Science and Pollution Research, Vol.9, No.5, pp.337-344.
Behnisch, P.A.; Hosoe, K.; Shiozaki, K.; Ozaki, H.; Nakamura, K.; Sakai, S., (2002c) Low-temperature thermal decomposition of dioxin-like compounds in fly ash: Combination of chemical analysis with in vitro bioassays (EROD and DR-CALUX). Environmental Science & Technology, Vol.36, No.23, pp.5211-5217.
Besselink, H.; Jones, A.; Pijnappels, M.; Swinkels, A.; Fjellanger, K.; Brouwer, A., (2003) Comparison of DR-CALUX® and HRGCMS-derived TEQ's: introduction of conversion factors. Organohalogen Compounds, Vol.60, pp.203-206.
Besselink, H.T.; Schipper, C.; Klamer, H.; Leonards, P.; Verhaar, H.; Felzel, E.; Murk, A.J.; Thain, J.; Hosoe, K.; Schoeters, G.; Legler, J.; Brouwer, B., (2004) Intra- and interlaboratory calibration of the DR Calux (R) bioassay for the analysis of dioxins and dioxin-like chemicals in sediments. Environmental Toxicology and Chemistry, Vol.23, No.12, pp.2781-2789.
Bosveld, A.T.C.; Berg, M.V., (1994) Biomarkers and bioassays as alternative screening methods for the presence and effects of PCDD, PCDF and PCB. Fresenius' Journal of Analytical Chemistry, Vol.348, No.1, pp.106-110.
Bradford, M.M., (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, Vol.72, No.1-2, pp.248-254.
Brouwer, A.; Reijnders, P.J.H.; Koeman, J.H., (1989) Polychlorinated biphenyl (PCB)-contaminated fish induces vitamin A and thyroid hormone deficiency in the common seal (Phoca vitulina). Aquatic Toxicology, Vol.15, No.1, pp.99-105.
Brown, D.J.; Chu, M.; Overmeire, I.V.; Chu, A.; Clark, G.C., (2001a) Determination of REP values for the CALUX bioassay and comparison to the WHO TEF values. Organohalogen Compounds, Vol.53, pp.211-214.
Brown, D.J.; Chu, M.; Van Overmeire, I.; Chu, A.; Clark, G.C., (2001b) Determination of REP values for the CALUX® bioassay and comparison to the WHO TEF values. Organohalogen Compounds, Vol.53, pp.211-214.
Brown, D.J.; Goeyens, L.; Overmeire, I.V.; Chu, M.; Murata, H.; Clark, G.C., (2001c) Quality control criteria implemented for monitoring the use of the CALUX bioassay. Organohalogen Compounds, Vol.54, pp.32-35.
Brown, D.J.; Orelien, J.; Gordon, J.D.; Chu, A.C.; Chu, M.D.; Nakamura, M.; Handa, H.; Kayama, F.; Denison, M.S.; Clark, G.C., (2007) Mathematical model developed for environmental samples: Prediction of GC/MS dioxin TEQ from XDS-CALUX bioassay data. Environmental Science & Technology, Vol.41, No.12, pp.4354-4360.
Brown, L.E.; Trought, K.R.; Bailey, C.I.; Clemons, J.H., (2005) 2,3,7,8-TCDD equivalence and mutagenic activity associated with PM10 from three urban locations in New Zealand. Science of the Total Environment, Vol.349, No.1-3, pp.161-174.
Burke, M.; Prough, R.; Mayer, R., (1977) Characteristics of a microsomal cytochrome P-448-mediated reaction. Ethoxyresorufin O-de-ethylation. Drug Metab Dispos, Vol.5, No.1, pp.1-8.
Burke, M.D.; Mayer, R.T., (1974) Ethoxyresorufin: direct fluorimetric assay of a microsomal O-dealkylation which is preferentially inducible by 3-methylcholanthrene. Drug Metabolism and Disposition, Vol.2, No.6, pp.583-588.
Carbonnelle, S.; Loco, J.V.; Overmeire, I.V.; Windal, I.; Wouwe, N.V.; Leeuwen, S.V.; Goeyens, L., (2004) Importance of REP values when comparing the CALUX bioassay results with chemoanalyses results: example with spiked vegetable oils. Talanta, Vol.63, No.5, pp.1255-1259.
Center, C.E.H.G.I., (2004) Food Storage: Refrigerator and Freezer. Food Safety, Nutrition, Diet & Health.
Chao, H.R.; Tsou, T.C.; Li, L.A.; Tsai, F.Y.; Wang, Y.F.; Tsai, C.H.; Chang, E.E.; Miao, Z.F.; Wu, C.H.; Lee, W.J., (2006) Arsenic inhibits induction of cytochrome P450 1A1 by 2,3,7,8-tetrachlorodibenzo-p-dioxin in human hepatoma cells. Journal of Hazardous Materials, Vol.137, No.2, pp.716-722.
Chao, H.R.; Wang, Y.F.; Chen, H.T.; Ko, Y.C.; Chang, E.E.; Huang, Y.J.; Tsai, F.Y.; Tsai, C.H.; Wu, C.H.; Tsou, T.C., (2007) Differential effect of arecoline on the endogenous dioxin-responsive cytochrome P450 1A1 and on a stably transfected dioxin-responsive element-driven reporter in human hepatoma cells. Journal of Hazardous Materials, Vol.149, pp.234-237.
Clemons, J.H.; Allan, L.M.; Marvin, C.H.; Wu, Z.; McCarry, B.E.; Bryant, D.W.; Zacharewski, T.R., (1998) Evidence of estrogen- and TCDD-like activities in crude and fractionated extracts of PM10 air particulate material using in vitro gene expression assays. Environmental Science & Technology, Vol.32, No.12, pp.1853-1860.
Dindal, A.; Thompson, E.; Aume, L.; Billets, S., (2007) Application of Site-Specific Calibration Data Using the CALUX by XDS Bioassay for Dioxin-like Chemicals in Soil and Sediment Samples. Environmental Science & Technology, Vol.41, No.24, pp.8376-8382.
Donato, M.T.; Gomezlechon, M.J.; Castell, J.V., (1993) A Microassay for Measuring Cytochrome P450IA1 and Cytochrome P450IIB1 Activities in Intact Human and Rat Hepatocytes Cultured on 96-Well Plates. Analytical Biochemistry, Vol.213, No.1, pp.29-33.
Engwall, M.; Hjelm, K., (2000) Uptake of dioxin-like compounds from sewage sludge into various plant species - assessment of levels using a sensitive bioassay. Chemosphere, Vol.40, No.9-11, pp.1189-1195.
European Commission, (2002) Commission Directive 2002/69/EC of 26 July 2002 laying down the sampling methods and the methods of analysis for the official control of dioxins and the determination of dioxin-like PCBs in foodstuffs Official journal of the European communities, Vol.L 209/5.
Gasiewicz, T.A.; Neal, R.A., (1982) The examination and quantitation of tissue cytosolic receptors for 2,3,7,8-tetrachlorodibenzo-p-dioxin using hydroxylapatite. Analytical Biochemistry, Vol.124, No.1, pp.1-11.
Goksøyr, A.; Husøy, A.-M., (1992) The cytochrome P450 1A1 response in fish: application of immunodetection in environmental monitoring and toxicological testing. Marine Environmental Research, Vol.34, No.1-4, pp.147-150.
Gonzalez-Doncel, M.; Fernandez-Torija, C.; Hinton, D.E.; Tarazona, J.V., (2005) Stage-specific toxicity of cypermethrin to medaka (Oryzias latipes) eggs and embryos using a refined methodology for an in vitro fertilization bioassay. Archives of Environmental Contamination and Toxicology, Vol.48, No.1, pp.87-98.
Gustavsson, L.; Hollert, H.; Jonsson, S.; van Bavel, B.; Engwall, M., (2007) Reed beds receiving industrial Sludge Containing Nitroaromatic compounds - Effects of outgoing water and bed material extracts in the umu-C genotoxicity assay, DR-CALUX assay and on early life stage development in Zebrafish (Danio rerio). Environmental Science and Pollution Research, Vol.14, No.3, pp.202-211.
Hahn, M.E., (2002) Biomarkers and bioassays for detecting dioxin-like compounds in the marine environment. The Science of The Total Environment, Vol.289, No.1-3, pp.49-69.
Hamers, T.; van Schaardenburg, M.D.; Felzel, E.C.; Murk, A.J.; Koeman, J.H., (2000) The application of reporter gene assays for the determination of the toxic potency of diffuse air pollution. Science of the Total Environment, Vol.262, No.1-2, pp.159-174.
Hankinson, O., (1995) The aryl hydrocarbon receptor complex. Annu. Rev. Pharmacol. Toxicol., Vol.35, pp.307-340.
Harper, P.A.; Golas, C.L.; Okey, A.B., (1988) Characterization of the Ah Receptor and Aryl Hydrocarbon Hydroxylase Induction by 2,3,7,8-Tetrachlorodibenzo-p-dioxin and Benz(a)anthracene in the Human A431 Squamous Cell Carcinoma Line. Cancer Res, Vol.48, No.9, pp.2388-2395.
Harrison, R.O.; Carlson, R.E., (1997) An immunoassay for TEQ screening of dioxin/furan samples: Current status of assay and applications development. Chemosphere, Vol.34, No.5-7, pp.915-928.
Hecker, M.; Murphy, M.B.; Giesy, J.P.; Hopkins, W.A., (2006) Induction of cytochrome P4501A in African brown house snake (Lamprophis fuliginosus) primary hepatocytes. Environmental Toxicology and Chemistry, Vol.25, No.2, pp.496-502.
Hertog, M.G.L.; Hollman, P.C.H.; Katan, M.B., (1992) Content of potentially anticarcinogenic flavonoids of 28 vegetables and 9 fruits commonly consumed in the Netherlands. Journal of Agricultural and Food Chemistry, Vol.40, No.12, pp.2379-2383.
Hertog, M.G.L.; Hollman, P.C.H.; van de Putte, B., (1993) Content of potentially anticarcinogenic flavonoids of tea infusions, wines, and fruit juices. Journal of Agricultural and Food Chemistry, Vol.41, No.8, pp.1242-1246.
Hilscherova, K.; Kannan, K.; Kang, Y.S.; Holoubek, I.; Machala, M.; Masunaga, S.; Nakanishi, J.; Giesy, J.P., (2001) Characterization of dioxin-like activity of sediments from a Czech river basin. Environmental Toxicology and Chemistry, Vol.20, No.12, pp.2768-2777.
Hilscherova, K.; Kannan, K.; Nakata, H.; Hanari, N.; Yamashita, N.; Bradley, P.W.; McCabe, J.M.; Taylor, A.B.; Giesy, J.P., (2003) Polychlorinated dibenzo-p-dioxin and dibenzofuran concentration profiles in sediments and flood-plain soils of the Tittabawassee River, Michigan. Environmental Science & Technology, Vol.37, No.3, pp.468-474.
Hinck, J.E.; Schmitt, C.J.; Blazer, V.S.; Denslow, N.D.; Bartish, T.M.; Anderson, P.J.; Coyle, J.J.; Dethloff, G.M.; Tillitt, D.E., (2006) Environmental contaminants and biomarker responses in fish from the Columbia River and its tributaries: Spatial and temporal trends. Science of the Total Environment, Vol.366, No.2-3, pp.549-578.
Hirobe, M.; Goda, Y.; Okayasu, Y.; Tomita, J.; Takigami, H.; Ike, M.; Tanaka, H., (2006) The use of enzyme-linked immunosorbent assays (ELISA) for the determination of pollutants in environmental and industrial wastes. Water Science and Technology, Vol.54, No.11-12, pp.1-9.
Hiyoshi Corporation (2006) Mechanism of CALUX cell.
Hollert, H.; Durr, M.; Olsman, H.; Halldin, K.; Van Bavel, B.; Brack, W.; Tysklind, M.; Engwall, M.; Braunbeck, T., (2002) Biological and chemical determination of dioxin-like compounds in sediments by means of a sediment triad approach in the catchment area of the River Neckar. Ecotoxicology, Vol.11, No.5, pp.323-336.
Hoogenboom, L.; Goeyens, L.; Carbonnelle, S.; van Loco, J.; Beernaert, H.; Baeyens, W.; Traag, W.; Bovee, T.; Jacobs, G.; Schoeters, G., (2006a) The CALUX bioassay: Current status of its application to screening food and feed. Trac-Trends in Analytical Chemistry, Vol.25, No.4, pp.410-420.
Hoogenboom, R., (2002) The combined use of the CALUX bioassay and the HRGC/HRMS method for the detection of novel dioxin sources and new dioxin-like compounds. Environmental Science and Pollution Research, Vol.9, No.5, pp.304-306.
Hoogenboom, R.; Bovee, T.; Traag, W.; Hoogerbrugge, R.; Baumann, B.; Portier, L.; van de Weg, G.; de Vries, J., (2006b) The use of the DR CALUX (R) bioassay and indicator polychlorinated biphenyls for screening of elevated levels of dioxins and dioxin-like polychlorinated biphenyls in eel. Molecular Nutrition & Food Research, Vol.50, No.10, pp.945-957.
Houtman, C.J.; Cenijn, P.H.; Hamers, T.; Lamoree, M.H.; Legler, J.; Murk, A.J.; Brouwer, A., (2004) Toxicological profiling of sediments using in vitro bioassays, with emphasis on endocrine disruption. Environmental Toxicology and Chemistry, Vol.23, No.1, pp.32-40.
Houtman, C.J.; Booij, P.; Jover, E.; del Rio, D.P.; Swart, K.; van Velzen, M.; Vreuls, R.; Legler, J.; Brouwer, A.; Lamoree, M.H., (2006) Estrogenic and dioxin-like compounds in sediment from Zierikzee harbour identified with CALUX assay-directed fractionation combined with one and two dimensional gas chromatography analyses. Chemosphere, Vol.65, No.11, pp.2244-2252.
Hurst, M.R.; Balaam, J.; Chan-Man, Y.L.; Thain, J.E.; Thomas, K.V., (2004) Determination of dioxin and dioxin-like compounds in sediments from UK estuaries using a bio-analytical approach: chemical-activated luciferase expression (CALUX) assay. Marine Pollution Bulletin, Vol.49, No.7-8, pp.648-658.
Hurst, M.R.; Chan-Man, Y.L.; Balaam, J.; Thain, J.E.; Thomas, K.V., (2005) The stable aryl hydrocarbon receptor agonist potency of United Kingdom Continental Shelf (UKCS) offshore produced water effluents. Marine Pollution Bulletin, Vol.50, No.12, pp.1694-1698.
Inouye, K.; Shinkyo, R.; Takita, T.; Ohta, M.; Sakaki, T., (2002) Metabolism of Polychlorinated Dibenzo-p-dioxins (PCDDs) by Human Cytochrome P450-Dependent Monooxygenase Systems. Journal of Agricultural and Food Chemistry, Vol.50, No.19, pp.5496-5502.
Iwamoto, K.; Kato, I.; Yu, X.M.; Nagasawa, S.; Kodaira, T.; Oishi, E.; Kitamura, K.; Usuki, Y.; Ohno, Y.; Yanaihara, N., (2004) Application of immunoassay to microquantitation of environmental endocrine disruptors - dioxins-specific immunoassay. Biomedical Research-Tokyo, Vol.25, No.1, pp.9-15.
JAPAN Ministry of the Environment Government, (2006) Bioassay Manual for Dioxin-like Compounds (Japanese). JAPAN Ministry of the Environment Government.
Joung, K.E.; Chung, Y.H.; Sheen, Y.Y., (2007) DRE-CALUX bioassay in comparison with HRGC/MS for measurement of toxic equivalence in environmental samples. Science of the Total Environment, Vol.372, No.2-3, pp.657-667.
Kammann, U.; Lang, T.; Vobach, M.; Wosniok, W., (2005) Ethoxyresorufin-O-deethylase (EROD) Activity in Dab (Limanda limanda) as Biomarker for Marine Monitoring. Environmental Science and Pollution Research, Vol.12, No.3, pp.140-145.
Kanematsu, M.; Shimizu, Y.; Sato, K.; Kim, S.; Suzuki, T.; Park, B.; Hattori, K.; Nakamura, M.; Yabushita, H.; Yokota, K., (2006) Distribution of dioxins in surface soils and river-mouth sediments and their relevance to watershed properties. Water Science and Technology, Vol.53, No.2, pp.11-21.
Kannan, K.; Villeneuve, D.L.; Yamashita, N.; Imagawa, T.; Hashimoto, S.; Miyazaki, A.; Giesy, J.P., (2000) Vertical profiles of dioxin-like and estrogenic activities associated with a sediment core from Tokyo Bay, Japan. Environmental Science & Technology, Vol.34, No.17, pp.3568-3573.
Katsamba, P.S.; Park, S.; Laird-Offringa, I.A., (2002) Kinetic studies of RNA-protein interactions using surface plasmon resonance. Methods, Vol.26, No.2, pp.95-104.
Klein, G.P.; Hodge, E.M.; Diamond, M.L.; Yip, A.; Dann, T.; Stem, G.; Denison, M.S.; Harper, P.A., (2006) Gas-phase ambient air contaminants exhibit significant dioxin-like and estrogen-like activity in vitro. Environmental Health Perspectives, Vol.114, No.5, pp.697-703.
Klotz, A.V.; Stegeman, J.J.; Walsh, C., (1984) An alternative 7-ethoxyresorufin o-deethylase activity assay: A continuous visible spectrophotometric method for measurement of cytochrome P-450 monooxygenase activity. Analytical Biochemistry, Vol.140, No.1, pp.138-145.
Koh, C.H.; Khim, J.S.; Villeneuve, D.L.; Kannan, K.; Giesy, J.P., (2006) Characterization of trace organic contaminants in marine sediment from Yeongil Bay, Korea: 2. Dioxin-like and estrogenic activities. Environmental Pollution, Vol.142, No.1, pp.48-57.
Koppen, G.; Covaci, A.; Van Cleuvenbergen, R.; Schepens, P.; Winneke, G.; Nelen, V.; Schoeters, G., (2001) Comparison of CALUX-TEQ values with PCB and PCDD/F measurements in human serum of the Flanders Environmental and Health Study (FLEHS). Toxicology Letters, Vol.123, No.1, pp.59-67.
Kopponen, P.; Törrönen, R.; Tarhanen, J.; Ruuskanen, J.; Kärenlampii, S., (1991) Cytochrome P450IA1 induction in mouse hepatoma cell culture as an indicator of polycyclic organic compounds in fly ash. Chemosphere, Vol.22, No.9-10, pp.895-904.
Kurosawa, S.; Aizawa, H.; Park, J.W., (2005) Quartz crystal microbalance immunosensor for highly sensitive 2,3,7,8-tetrachlorodibenzo-p-dioxin detection in fly ash from municipal solid waste incinerators. Analyst, Vol.130, No.11, pp.1495-1501.
Kurosawa, S.; Park, J.-W.; Aizawa, H.; Wakida, S.-I.; Tao, H.; Ishihara, K., (2006) Quartz crystal microbalance immunosensors for environmental monitoring. Biosensors and Bioelectronics, Vol.22, No.4, pp.473-481.
Laier, P.; Cederberg, T.; Larsen, J.C.; Vinggaard, A.M., (2003) Applicability of the CALUX bioassay for screening of dioxin levels in human milk samples. Food Additives and Contaminants, Vol.20, No.6, pp.583-595.
Leskinen, P.; Hilscherova, K.; Sidlova, T.; Kiviranta, H.; Pessala, P.; Salo, S.; Verta, M.; Virta, M., (2008) Detecting AhR ligands in sediments using bioluminescent reporter yeast. Biosensors and Bioelectronics, Vol.23, No.12, pp.1850-1855.
Li, W.; Wu, W.Z.; Barbara, R.B.; Schramm, K.W.; Kettrup, A., (1999) A new enzyme immunoassay for PCDD/F TEQ screening in environmental samples: comparison to micro-EROD assay and to chemical analysis. Chemosphere, Vol.38, No.14, pp.3313-3318.
Li, W.; Wu, W.Z.; Xu, Y.; Li, L.; Schramm, K.W.; Kettrup, A., (2002) Measuring TCDD Equivalents in Environmental Samples with the Micro-EROD Assay: Comparison with HRGC/HRMS Data. Bulletin of Environmental Contamination and Toxicology, Vol.68, No.1, pp.111-117.
Liu, X.; Song, D.; Zhang, Q.; Tian, Y.; Zhang, H., (2004) An optical surface plasmon resonance biosensor for determination of tetanus toxin. Talanta, Vol.62, No.4, pp.773-779.
Ma, M.; Li, J.; Wang, Z.J., (2005) Assessing the detoxication efficiencies of wastewater treatment processes using a battery of bioassays/biomarkers. Archives of Environmental Contamination and Toxicology, Vol.49, No.4, pp.480-487.
Machala, M.; Vondracek, J.; Blaha, L.; Ciganek, M.; Neca, J., (2001) Aryl hydrocarbon receptor-mediated activity of mutagenic polycyclic aromatic hydrocarbons determined using in vitro reporter gene assay. Mutation Research-Genetic Toxicology and Environmental Mutagenesis, Vol.497, No.1-2, pp.49-62.
Mandal, P.K., (2005) Dioxin: a review of its environmental effects and its aryl hydrocarbon receptor biology. Journal of Comparative Physiology B: Biochemical, Systemic, and Environmental Physiology, Vol.175, No.4, pp.221-230.
McAlister, D.R.; Fern, M.J.; Allen, R.L., (2008) Rapid aryl hydrocarbon receptor based polymerase chain reaction screening assay for polychlorinated dibenzo-p-dioxins and furans in soil and sediment. Talanta, Vol.74, No.4, pp.992-997.
Mohanty, S.P.; Kougianos, E., (2006) Biosensors: a tutorial review. IEEE POTENTIALS, Vol.25, pp.35-40
Montgomery, D.C., (2001). Statistical basis of the control chart. Introduction to statistical quality control John Wiley, New York.
Murk, A.J.; Legler, J.; Denison, M.S.; Giesy, J.P.; vandeGuchte, C.; Brouwer, A., (1996) Chemical-activated luciferase gene expression (CALUX): A novel in vitro bioassay for Ah receptor active compounds in sediments and pore water. Fundamental and Applied Toxicology, Vol.33, No.1, pp.149-160.
Murk, A.J.; Leonards, P.E.G.; Bulder, A.S.; Jonas, A.S.; Rozemeijer, M.J.C.; Denison, M.S.; Koeman, J.H.; Brouwer, A., (1997) The CALUX (chemical-activated luciferase expression) assay adapted and validated for measuring TCDD equivalents in blood plasma. Environmental Toxicology and Chemistry, Vol.16, No.8, pp.1583-1589.
Murphy, M.B.; Hecker, M.; Coady, K.K.; Tompsett, A.R.; Jones, P.D.; Newsted, J.L.; Wong, H.L.; du Preez, L.H.; Solomon, K.R.; Carr, J.A.; Smith, E.E.; Kendall, R.J.; Van der Kraak, G.; Giesy, J.P., (2006) Sediment TCDD-EQs and EROD and MROD activities in ranid frogs from agricultural and nonagricultural sites in Michigan (USA). Archives of Environmental Contamination and Toxicology, Vol.51, No.3, pp.467-477.
Nagy, S.R.; Liu, G.; Lam, K.S.; Denison, M.S., (2002a) Identification of novel Ah receptor agonists using a high-throughput green fluorescent protein-based recombinant cell bioassay. Biochemistry, Vol.41, No.3, pp.861-868.
Nagy, S.R.; Sanborn, J.R.; Hammock, B.D.; Denison, M.S., (2002b) Development of a green fluorescent protein-based cell Bioassay for the rapid and inexpensive detection and characterization of Ah receptor agonists. Toxicological Sciences, Vol.65, No.2, pp.200-210.
Nebert, D.W.; Gelboin, H.V., (1968) Substrate-inducible microsomal aryl hydroxylase in mammalian cell culture. I. assay and properties of induced enzyme. J. Biol. Chem., Vol.243, No.23, pp.6242-6249.
Nichkova, M.; Park, E.K.; Koivunen, M.E.; Kamita, S.G.; Gee, S.J.; Chuang, J.; Van Emon, J.M.; Hammock, B.D., (2004) Immunochemical determination of dioxins in sediment and serum samples. Talanta, Vol.63, No.5, pp.1213-1223.
Nording, M.; Denison, M.S.; Baston, D.; Persson, Y.; Spinnel, E.; Haglund, P., (2007) 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, Vol.26, No.6, pp.1122-1129.
Nunes, G.S.; Barcel, D.; Grabaric, B.S.; Díaz-Cruz, J.M.; Ribeiro, M.L., (1999) Evaluation of a highly sensitive amperometric biosensor with low cholinesterase charge immobilized on a chemically modified carbon paste electrode for trace determination of carbamates in fruit, vegetable and water samples. Analytica Chimica Acta, Vol.399, No.1-2, pp.37-49.
Orrego, R.; Jiménez, B.; Bordajandi, L.R.; Gavilán, J.F.; Inzunza, B.; Abad, E.; González, M.J.; Rivera, J.; Barra, R., (2005) EROD induction and PCDD/F levels in fish liver from the Biobio River in Chile. Chemosphere, Vol.60, No.7, pp.829-835.
Overmeire, I.V.; Loco, J.V.; Roos, P.; Carbonnelle, S.; Goeyens, L., (2004) Interpretation of CALUX results in view of the EU maximal TEQ level in milk. Talanta, Vol.63, pp.1241-1247.
Pohl, R.J.; Fouts, J.R., (1980) A rapid method for assaying the metabolism of 7-ethoxyresorufin by microsomal subcellular fractions. Analytical Biochemistry, Vol.107, No.1, pp.150-155.
Poland, A.; Glover, E., (1975) Genetic expression of aryl hydrocarbon hydroxylase by 2,3,7,8-tetrachlorodibenzo-p-dioxin: evidence for a receptor mutation in genetically non-responsive mice. Molecular Pharmacology, Vol.11, pp.389-398.
Rogers, K.R., (2006) Recent advances in biosensor techniques for environmental monitoring. Analytica Chimica Acta, Vol.568, No.1-2, pp.222-231.
Roy, S.; Mysior, P.; Brzezinski, R., (2002) Comparison of dioxin and furan TEQ determination in contaminated soil using chemical, micro-EROD, and immunoassay analysis. Chemosphere, Vol.48, No.8, pp.833-842.
Safe, S.; Bandiera, S.; Sawyer, T.; Zmudzka, B.; Mason, G.; Romkes, M.; Denomme, M.A.; Sparling, J.; Okey, A.B.; Fujita, T., (1985) Effects of structure on binding to the 2,3,7,8-TCDD receptor protein and AHH induction--halogenated biphenyls. Environ Health Perspect, Vol.61, pp.21-33.
Safe, S.H., (1995) Modulation of gene expression and endocrine response pathways by 2,3,7,8-tetrachlorodibenzo-p-dioxin and related compounds. Pharmacology & Therapeutics, Vol.67, No.2, pp.247-281.
Saito, K.; Ishizuka, M.; Hosono, S.; Sugawara, Y.; Iwasaki, Y.; Ito, R.; Nakazawa, H., (2006) Development of screening method of dioxins in environmental samples by enzyme-linked immunosorbent assay. Bunseki Kagaku, Vol.55, No.12, pp.993-997.
Sanborn, J.R.; Gee, S.J.; Gilman, S.D.; Sugawara, Y.; Jones, A.D.; Rogers, J.; Szurdoki, F.; Stanker, L.H.; Stoutamire, D.W.; Hammock, B.D., (1998) Hapten synthesis and antibody development for polychlorinated dibenzo-p-dioxin immunoassays. Journal of Agricultural and Food Chemistry, Vol.46, No.6, pp.2407-2416.
Sanctorum, H.; Windal, I.; Hanot, V.; Goeyens, L.; Baeyens, W., (2007) Dioxin and dioxin-like activity in sediments of the belgian coastal area (Southern north sea). Archives of Environmental Contamination and Toxicology, Vol.52, No.3, pp.317-325.
Schneider, U.A.; Brown, M.M.; Logan, R.A.; Millar, L.C.; Bunce, N.J., (1995) Screening assay for dioxin-like compounds based on competitive binding to the murine hepatic Ah receptor. 1. Assay development. Environmental Science & Technology, Vol.29, No.10, pp.2595-2602.
Schroijen, C.; Windal, I.; Goeyens, L.; Baeyens, W., (2004) Study of the interference problems of dioxin-like chemicals with the bio-analytical method CALUX. Talanta, Vol.63, No.5, pp.1261-1268.
Schwirzer, S.M.G.; Hofmaier, A.M.; Kettrup, A.; Nerdinger, P.E.; Schramm, K.W.; Thoma, H.; Wegenke, M.; Wiebel, F.J., (1998) Establishment of a simple cleanup procedure and bioassay for determining 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicity equivalents of environmental samples. Ecotoxicology and Environmental Safety, Vol.41, No.1, pp.77-82.
Scippo, M.L.; Eppe, G.; De Pauw, E.; Maghuin-Rogister, G., (2004) DR-CALUX((R)) screening of food samples: evaluation of the quantitative approach to measure dioxin, furans and dioxin-like PCBs. Talanta, Vol.63, No.5, pp.1193-1202.
Shan, G.; Leeman, W.R.; Gee, S.J.; Sanborn, J.R.; Jones, A.D.; Chang, D.P.Y.; Hammock, B.D., (2001) Highly sensitive dioxin immunoassay and its application to soil and biota samples. Analytica Chimica Acta, Vol.444, No.1, pp.169-178.
Shelver, W.L.; Shan, G.; Gee, S.J.; Stanker, L.H.; Hammock, B.D., (2002) Comparison of immunoaffinity column recovery patterns of polychlorinated dibenzo-p-dioxins/polychlorinated dibenzofurans on columns generated with different monoclonal antibody clones and polyclonal antibodies. Analytica Chimica Acta, Vol.457, No.2, pp.199-209.
Shimomura, M.; Nomura, Y.; Zhang, W.; Sakino, M.; Lee, K.-H.; Ikebukuro, K.; Karube, I., (2001) Simple and rapid detection method using surface plasmon resonance for dioxins, polychlorinated biphenylx and atrazine. Analytica Chimica Acta, Vol.434, No.2, pp.223-230.
Singer, R.; Lansky, D.M.; Hauck, W.W., (2006) Bioassay glossary. Pharmacopeial Forum, Vol.32, No.4, pp.1359-1365.
Skarek, M.; Janosek, J.; Cupr, P.; Kohoutek, J.; Novotna-Rychetska, A.; Holoubek, I., (2007) Evaluation of genotoxic and non-genotoxic effects of organic air pollution using in vitro bioassays. Environment International, Vol.33, pp.859-866.
Song, C.L.; Huang, R.J.; Wang, Y.Q.; Liu, K.M.; Dong, S.R.; Dai, S.G., (2006a) Determination of in vitro biotoxicity in exhaust particulate matter from heavy-duty diesel engine. Bulletin of Environmental Contamination and Toxicology, Vol.76, No.1, pp.24-32.
Song, M.Y.; Jiang, Q.T.; Xu, Y.; Liu, H.X.; Lam, P.K.S.; O'Toole, D.K.; Zhang, Q.H.; Giesy, J.P.; Jiang, G.B., (2006b) AhR-active compounds in sediments of the Haihe and Dagu Rivers, China. Chemosphere, Vol.63, No.7, pp.1222-1230.
Sugawara, Y.; Gee, S.J.; Sanborn, J.R.; Gilman, S.D.; Hammock, B.D., (1998) Development of a highly sensitive enzyme-linked immunosorbent assay based on polyclonal antibodies for the detection of polychlorinated dibenzo-p-dioxins. Analytical Chemistry, Vol.70, No.6, pp.1092-1099.
Suzuki, G.; Takigami, H.; Kushi, Y.; Sakai, S.-i., (2004a) Evaluation of mixture effects in a crude extract of compost using the CALUX bioassay and HPLC fractionation. Environment International, Vol.30, No.8, pp.1055-1066.
Suzuki, G.; Takigami, H.; Kushi, Y.; Sakai, S., (2004b) Evaluation of mixture effects in a crude extract of compost using the CALUX bioassay and HPLC fractionation. Environment International, Vol.30, No.8, pp.1055-1066.
Taiwan Environmental Protection Administration, (2005) Analysis method of dioxins and furans: isotope dilution high resolution gas chromatography-mass spectrometry (Chinese), NIEA M801.11B.
Tsutsumi, T.; Amakura, Y.; Nakamura, M.; Brown, D.J.; Clark, G.C.; Sasaki, K.; Toyoda, M.; Maitani, T., (2003) Validation of the CALUX bioassay for the screening of PCDD/Fs and dioxin-like PCBs in retail fish. Analyst, Vol.128, No.5, pp.486-492.
Tsutsumi, T.; Miyoshi, N.; Sasaki, K.; Maitani, T., (2008) Biosensor immunoassay for the screening of dioxin-like polychlorinated biphenyls in retail fish. Analytica Chimica Acta, Vol.617, No.1-2, pp.177-183.
U.S. EPA, (1994) Tetra- through Octa Chlorinated Dioxins and Furans by Isotope Dilution HRGC/HRMS. In: U.S. Environmental Protection Agency (Ed.).
U.S. EPA, (2005) Technologies for monitoring and measurement of dioxin and dioxin-like compounds in soil and sediment. In: U.S. EPA office and research and development (Ed.). Innovative Technology Verification Report. USEPA office and research and development.
U.S. EPA, (2008) Method for toxic equivalents (TEQs) determinations for dioxin-like chemical activity with the CALUX® bioassay. U.S. EPA.
U.S.EPA, (2007a) Method 8280B polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) by high resolution gas chromatography/low resolution mass spectrometry (HRGC/LRMS). U.S. Environmental Protection Agency.
U.S.EPA, (2007b) Method 8290A polychlorinated dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) by high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS). U.S. Environmental Protection Agency.
van den Heuvel, M.R.; Munkittrick, K.R.; Stegeman, J.J.; Dixon, D.G., (1995) Second-round interlaboratory comparison of hepatic ethoxyresorufin-o-deethylase activity in white sucker (catostomus commersoni) exposed to bleached-kraft pulp mill effluent. Environmental Toxicology and Chemistry, Vol.14, No.9, pp.1513-1520.
van der Oost, R.; Beyer, J.; Vermeulen, N.P.E., (2003) Fish bioaccumulation and biomarkers in environmental risk assessment: a review. Environmental Toxicology and Pharmacology, Vol.13, No.2, pp.57-149.
Van Wouwe, N.; Windal, I.; Vanderperren, H.; Eppe, G.; Xhrouet, C.; Massart, A.C.; Debacker, N.; Sasse, A.; Baeyens, W.; De Pauw, E.; Sartor, F.; Van Oyen, H.; Goeyens, L., (2004) Validation of the CALUX bioassay for PCDD/F analyses in human blood plasma and comparison with GC-HRMS. Talanta, Vol.63, No.5, pp.1157-1167.
Vanderlaan, M.; Stanker, L.H.; Watkins, B.E.; Petrovic, P.; Gorbach, S., (1988a) Improvement and application of an immunoassay for screening environmental samples for dioxin contamination. Environmental Toxicology and Chemistry, Vol.7, No.11, pp.859-870.
Vanderlaan, M.; Watkins, B.E.; Stanker, L., (1988b) ES&T critical review: environmental monitoring by immunoassay. Environmental Science & Technology, Vol.22, No.3, pp.247-254.
Vanderperren, H.; Van Wouwe, N.; Behets, S.; Windal, I.; Van Overmeire, I.; Fontaine, A., (2004) TEQ-value determinations of animal feed; emphasis on the CALUX bioassay validation. Talanta, Vol.63, No.5, pp.1277-1280.
Wang, W.; Gao, X.B.; Zheng, L.; Lan, Y.X., (2006) Reductive dechlorination of polychlorinated dibenzo-p-dioxins and dibenzofurans in MSWI fly ash by sodium hypophosphite. Separation and Purification Technology, Vol.52, No.1, pp.186-190.
Warner, M.; Eskenazi, B.; Patterson, D.G.; Clark, G.; Turner, W.E.; Bonsignore, L.; Mocarelli, P.; Gerthoux, P.M., (2005) Dioxin-like TEQ of women from the Seveso, Italy area by ID-HRGC/HRMS and CALUX. Journal of Exposure Analysis and Environmental Epidemiology, Vol.15, No.4, pp.310-318.
Watkins, B.E.; Stanker, L.H.; Vanderlaan, M., (1989) An immunoassay for chlorinated dioxins in soils. Chemosphere, Vol.19, No.1-6, pp.267-270.
Weiden, M.E.J.v.d.; Celander, M.; Seinen, W.; Berg, M.v.d.; Goksøyr, A.; Förlin, L., (1993) Induction of cytochrome P450 1A in fish treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin or chemically contaminated sediment. Environmental Toxicology and Chemistry, Vol.12, pp.989-999.
Windal, I.; Van Wouwe, N.; Eppe, G.; Xhrouet, C.; Debacker, V.; Baeyens, W.; De Pauw, E.; Goeyens, L., (2005) Validation and interpretation of CALUX as a tool for the estimation of dioxin-like activity in marine biological matrixes. Environmental Science & Technology, Vol.39, No.6, pp.1741-1748.
Wouwe, N.V.; Windal, I.; Vanderperren, H.; Eppe, G.; Xhrouet, C.; Massart, A.-C.; Debacker, N.; Sasse, A.; Baeyens, W.; Pauw, E.D.; Sartor, F.; Oyen, H.V.; Goeyens, L., (2004) Validation of the CALUX bioassay for PCDD/F analyses in human blood plasma and comparison with GC-HRMS. Talanta, Vol.63, pp.1157-1167.
Xenobiotic Detection Systems, (2005a) sample preparation training manual. Xenobiotic Detection Systems, Inc. .
Xenobiotic Detection Systems, (2005b) cell culture training manual. Xenobiotic Detection Systems, Inc.
Yan, J.H.; Chen, T.; Li, X.D.; Zhang, J.; Lu, S.Y.; Ni, M.J.; Cen, K.F., (2006) Evaluation of PCDD/Fs emission from fluidized bed incinerators co-firing MSW with coal in China. Journal of Hazardous Materials, Vol.135, No.1-3, pp.47-51.
Zhou, X.C.; Cao, L., (2001) High sensitivity microgravimetric biosensor far qualitative and quantitative diagnostic detection of polychlorinated dibenzo-p-dioxins. Analyst, Vol.126, No.1, pp.71-78.