行政院環境保護署 環保專案成果報告查詢系統

EPA-98-1604-02-01

098

1,470 千元

2009/05/26

2010/05/25

鄒粹軍副研究員

環檢所

陳元武研究員

財團法人國家衛生研究院

環境監測及檢測

國立屏東科技大學 環境工程與科學系 趙浩然副教授;中原大學 生物環境工程學系 王雅玢副教授

鄒粹軍副研究員

037-246166#36511

tctsou@nhri.org.tw

國內焚化爐所產生之戴奧辛污染，是國人極為關切之環境議題，因此，工廠本身每年需定期進行焚化爐之煙道氣戴奧辛監測；而環保單位則需進行稽查工作，故其分析所耗的時間及經濟成本相當龐大。戴奧辛對於自然環境及人類健康之毒害日益受到重視，戴奧辛造成的毒性反應範圍相當廣泛，大致上可分為皮膚毒性、肝臟毒性、免疫毒性、生殖毒性與致癌性等，當戴奧辛透過食物鏈進入人體中，會產生類似荷爾蒙的作用而形成「假性荷爾蒙」效應，由此干擾內分泌系統的調控；一般而言，檢測戴奧辛的方法以高解析度氣相層析與高解析度質譜儀（HRGC/HRMS）為主，由於此項化學分析技術所需之成本甚高且耗時過長，在分析樣品日益增加的現代，建立快速、有效且低成本的檢測方法 (如生物檢測) 有其迫切性。目前，本實驗室已成功開發出戴奧辛/類戴奧辛生物檢測法(chemical-activated luciferase expression bioassay, 簡稱CALUX)之細胞株，此生物檢測方法的原理，是利用戴奧辛/類戴奧物質能夠活化細胞內芳香族羥基碳氫化合物接受器 (Aryl hydrocarbon receptor，簡稱AhR) 之特性，量測經由AhR所誘發之luciferase酵素活性，由此可推估出檢測樣品中戴奧辛/類戴奧物質之總含量，由於此特性，DRE-luciferase bioassay之檢測範圍，並不局限於某些特定或單一的戴奧辛/類戴奧物質。本計畫擬以多方合作的方式，以本實驗室開發之DRE-luciferase bioassay為基礎，由屏東科技大學新興污染物中心（趙浩然博士）及中原大學生物環境工程系（王雅玢博士）負責研發快速有效之前處理方法，對焚化爐所產生之煙道氣樣品，進行戴奧辛/類戴奧物質總量之檢測，所得生物檢測數據將與HRGC/HRMS所測得之數據進行比對，藉以探討此生物檢測法對煙道氣樣品之適用性，以期開發快速、有效而低成本的本土戴奧辛生物篩檢技術，並推廣至環境檢測機構及學術單位，此研究結果將有效降低執行戴奧辛監測之時間及經濟成本。

環檢所煙道氣生物偵測計畫期末報告.pdf

24MB

環檢所煙道氣生物偵測計畫期末報告

2010/7/1 上午 12:00:00

Recent techniques for detection and quantification of dioxins and dioxin-like compounds involve mainly the costly and time-consuming high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS). To ensure environmental safety, it is extremely crucial to develop high-throughput assays for screening potential contamination of dioxin-like chemicals in various field samples from environments and foods. The Chemical Activated LUciferase eXpression (CALUX) bioassay is an in vitro luciferase-reporter-gene assay for detecting the Toxic Equivalents (TEQ) levels of dioxins and dioxin-like compounds based on their high affinity with aryl hydrocarbon receptor (AhR). As compared with HRGC/HRMS, CALUX bioassay is a fast and low-cost method for high-throughput screening of those highly dioxin-contaminated samples from environment and foods. The technique relies on selection of stable dioxin-responsive cell lines that can maintain their responsiveness to dioxins and dioxin-like compounds. We have established a stable dioxin-responsive cell line, Huh7-DRE-Luc. In the present study, we have determined the capability of this cell line (DRE-luciferase bioassay) in detection of dioxins and dioxin-like compounds by testing with 2,3,7,8-TCDD, CRM powder, EDF-5416, and samples from various matrices (i.e., soil, sediment, and flue gas). Our results showed that the ratio of DRE-Luciferase reading (BIO) to HRGC/HRMS reading (CHE) in testing with 2,3,7,8-TCDD was around 1:1, the ratio of BIO to CHE in testing with CRM powder was around 5:1, and the ratio of BIO to CHE in testing with EDF-5416 was around 3.9:1. Meanwhile, the BIO/CHE ratios in testing with samples from soil, sediment, and flue gas are 31.2, 21.7 and 26.7, respectively. Regarding testing with pure 2,3,7,8-TCDD (with BIO/CHE ratios around 1 and TCDD detection limit of 35.4 pM), Huh7-DRE-Luc is suggested to be a qualified system for detection of dioxins and dioxin-like compounds. However when pre-treatment steps are included in the sample preparation, the BIO/CHE ratios raise markedly. This data suggests that our pre-treatment procedure may interfere with the BIO/CHE ratios. On the basis of these results, the objectives of our future studies are to improve the TCDD detection sensitivity of our DRE-luciferase bioassay system and to find out the factor(s) that may affect the BIO/CHE ratios during the pre-treatment of samples.