- 學位論文
戴奧辛冷光酵素基因表現法應用於飛灰樣品之研究
Application of DRE-Luciferase bioassay on ash samples
摘要
戴奧辛和呋喃，持久性有機污染物高安定性與低分解性，具有生物累積和放大作用，在環境中容易累積於食物鏈中最後進入人體，因其高親脂性而積存於脂肪中，對人類健康會造成深遠的威脅。 目前分析戴奧辛類化合物主要是藉由HRGC-HRMS化學方法進行，然而戴奧辛污染事件通常為緊急突發事件，需要在短時間內進行大量樣品篩檢，化學分析雖然具備精準之分析結果，但檢測過程耗時費事、成本昂貴，無法符合緊急應變事件之快速要求，因此國際上已發展戴奧辛快速生物篩檢技術，降低分析成本並且能於短時間內篩檢大量樣品。 本研究利用三種萃取溶劑 (正己烷/二氯甲烷、正己烷、甲苯) 輔以兩種萃取方法 (超音波萃取及索氏萃取) 及兩種淨化程序 (直接濃縮及管柱淨化) 來處理飛灰，並以戴奧辛冷光酵素基因表現法來分析樣品間戴奧辛含量之差異，結果顯示，墊基於大鼠肝腫瘤細胞株 (Ad-DRE-Luc/H4IIE)所發展出之戴奧辛冷光酵素基因表現法對於樣品經由超音波萃取之管柱淨化程序結果具有良好的相關性，在生物分析與化學分析結果之三種溶劑共同比較下其相關係數為0.873，其中以甲苯為萃取溶劑相關性最好 (R2=0.984) ，其次是正己烷/二氯甲烷 (R2=0.929)，最後是正己烷 (R2=0.883) ；在索氏萃取管柱淨化方面，正己烷/二氯甲烷與甲苯溶劑萃取也具有不錯之相關性，分別為R2=0.898及R2=0.887；兩種萃取方法在樣品直接濃縮上，生物分析與化學分析之相關性結果遠低於管柱淨化，超音波萃取直接濃縮與索氏萃取直接濃縮之相關係數分別為0.010、-0.026， 研究初步結果顯示，戴奧辛冷光酵素基因表現法對於飛灰管柱淨化後之樣品具有較良好的檢測效果。
並列摘要
Dioxin and Furan (simply as Dioxins) are persistent organic pollutants which are highly stable and less decomposed. Dioxins are bioaccumulated and biomagnified. They are easily accumulated in the food chain and finally enter the human bodies to result in health impact. Therefore, fast screen these dioxins-polluted foods is a hot issue. HRGC-HRMS is the main chemical analysis method for dioxins. The dioxin pollution events are always breaking news and require instant response to provide valuable information and data within a short time. Chemical analysis method is though accurate, but time-consuming and expensive, not suitable for largescale dioxin surveillance. Therefore, Chemically Activated LUciferase eXpression bioassay was progressively developed to apply to the fast prescreening. In this study, three extracted solvent: toluene, n-hexane/methylene chloride and n-hexane; two extracted method: soxhlet extraction and ultrasonic extraction and two clean up procedure: direct concentration and column clean up were applied to pretreat ash samples. Ad-DRE-Luc/H4IIE cells based DRE-Luciferase bioassay was used for analysis. The results showed that there was good correlation between DRE-Luciferase bioassay and chemical analysis for ultrasonic extracted samples following column clean up procedure. The correlation coefficient is 0.873 for all three solvent samples. Toluene (R2=0.984) as the extraction solvent had the best outcome, followed by n-hexane/Methylene chloride (R2=0.929) and n-hexane (R2=0.883). For soxhlet extraction column clean up process, n-hexane/Methylene chloride and the toluene solvent extracts also had good correlation, R2=0. 898 and R2=0.887, respectively. The low correlation coefficient of DRE-Luciferase bioassay and chemical analysis method for direct concentration samples between ultrasonic extraction and soxhlet extraction was only 0.010 and -0.026. The preliminary result shows that DRE-Luciferase bioassay following the column clean up pretreatment provided a better detection results for ash samples.