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年度	106	專案性質	實驗性質
專案類別	研究專案	研究主題	評估
申請機構	國立屏東科技大學	申請系所	環境工程與科學系
專案主持人	趙浩然	職等/職稱	教授
專案中文名稱	發展毒性監測及預報平台-使用高通量生物篩選工具預測土壤中環境荷爾蒙污染物
中文關鍵字	高通量,環境荷爾蒙,體外試驗(in-vitro),毒性監測
專案英文名稱	Developing toxicity monitor and prediction - use of high-throughput bio-screening assays to forecast environmental hormones in soils
英文關鍵字	High-throughput, Environmental hormone, in-vitro, toxic monitoring
執行金額	900,000元
執行期間	2017/1/9 至 2017/11/30
計畫中文摘要	美國環保署於2012年開始運作Endocrine Disruptor Screening Program (EDSP)，計劃篩選85,000種新化合物確認是否有內分泌干擾效應，達成毒性預報的目標。生物分析法為一種被廣泛應用於污染物之篩檢工具，其快速且操作容易，可分析複雜化學混合物中的總毒性效應。因此，本研究目的為建立高通量篩選工具(HTS assays)，包含致癌性篩選平台(p53 bioassay)、綜合毒性篩選平台(ATP biosassay)、芳香族化合物毒性篩選平台(Ad-DR bioassay)及雌激素毒性篩選平台(T47D-KBluc assay)，可分析土樣品中環境化學品之毒性，並進一步進行風險評估。本研究土壤樣品位於高雄市某受重油品污染之區域，已完成30件次化學法分析與140件次四種HTS分析測定。本研究結果顯示，污染程度較高之土壤樣本其有機相平均細胞存活率為83%，顯著低於其他三種土壤樣本(低污染、輕微污染與背景土樣)，且與BTEX化合物均呈現負相關，但未達顯著；無論在有機相或無機相受污染土壤樣本所誘發之DNA損傷顯著高於背景土樣(p=0.002)；戴奧辛與Non-Dioxin AhR濃度範圍分別為ND~1.50 pg BEQ/g d.w.與ND~29.7 pg BEQ/g d.w.，3種BTEX(苯、乙本與二甲苯)均與Non-Dioxin AhR數據呈現顯著正相關(p<0.05)，在TPH方面則有邊緣顯著之現象(p=0.071)；僅於有機相中檢測出微量之雌激素效應，部分樣本其濃度均低於本研究偵測極限。本研究已建立毒性預報系統流程圖，並遵循污染物指紋鑑定之概念，建立受油品污染土樣之生物指紋圖，後續應用本系統針對未知樣本測試時，可快速評估該地是否遭受油品污染之情形。
計畫英文摘要	The Environmental Protection Agency (EPA) began the implementation of Endocrine Disruptor Screening Program (EDSP) in 2012 which aims to screen 85,000 new compounds for potential endocrine disrupting effects and tox-cast targets. Bioassays are becoming increasingly attractive screening tools due to the relative speed and ease of analysis, and can provide an integrated measure of toxicity potency in complex chemical mixtures. Therefore, the aim of this study is to establish high-throughput screening assays (HTS assays) including the carcinogenicity pathway (p53 bioassay), cytotoxicity (ATP biosassay), aryl hydrocarbon receptor pathway (Ad-DR bioassay) and the estrogen receptor pathway (T47D-kbLuc assay) to screen for toxicity of environmental chemicals and to do further risk assessment in soil samples. Thirty (30) and One hundred forty (140) samples were analyzed on GC/FID and four HTS assays, respectively which were located in a heavy oil pollutant area in Kaohsiung city. Compared to other samples in our study, our results showed low cell survival rate (83%) in samples heavily polluted with oil. In addition, Negative correlations of cell survival rate and BTEX pollutants were observed in whole samples. Significant increase in DNA induced damage were incurred in pollution soils either in the organic or inorganic extraction fractions. The range of dioxin andNon-Dioxin AhR luciferase responses are ND~1.50 pg BEQ/g d.w and ND~29.7 pg BEQ/g d.w. which was observed with positive correlation and marginal positive correlation with three BTEX compounds namely: benzene, ethyl benzene and dimethylbenzene and TPH. Notably, very low estrogenic activity was observed in organic fraction and most sample concentrations were lower than limit of detection. In conclusion, the toxicity report system based on four HTS assays have been established and to be able to make a bioassay fingerprinting of soil heavily contaminated with oil to further assess and evaluate on unknown soil samples.