Application of Bioassay on Enviromental Samples

• Main Authors: Yen-Sheng You, 游彥勝
• Other Authors: Ya-Fen Wang
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/87371192740411296025

碩士 === 中原大學 === 土木工程研究所 === 98 === Abstract Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are polychlorinated aromatic compounds that are accumulated mainly in fatty tissues due to their high lipophilicity, long biological half-lives, and high resistance to biodegradation. 2,3,7,8 - TCDD, the most toxic PCDD/Fs, was classified as carcinogenic to humans (Group 1). General population exposure to PCDD/DFs can induce several adverse health effects including reproductive, hormonal, immunological, and carcinogenic effects . High-resolution gas chromatography / high-resolution mass spectrometry (HRGC/HRMS) is a certified method to determine dioxin concentrations in Taiwan. However, it costs a lot and takes a lot of time. The chemical activated luciferase expression bioassay has been recently established and showed potential to analyze PCDD/Fs. The bioassay is an in vitro luciferase-reporter gene assay for detecting the TEQ levels of dioxins and dioxin-like compounds based on their high affinity with aryl hydrocarbon receptor (AhR). Compared with HRGC/HRMS, the bioassay is a rapid and low-cost screening method particularly for fast-screen identification of highly dioxin-contaminated samples from environment and food. However, the commercial cell-line is too expensive and hard to be popularized to industry. In this study, a stably recombinant cell-line, Huh7-DRE-Luc cell, was established. The capability of Huh7-DRE-Luc cell (DRE-Luciferase assay) was tested by 2,3,7,8-TCDD standard, CRM powder, sediments, soils and flue gas samples. The results indicated that the ratios of DRE-Luciferase assay (BIO) to HRGC/HRMS (CHE) for solid and liquid standards were 5.48 and 3.98, respectively. When 2,3,7,8-TCDD concentration was greater than 78.7 pM, the reliability of dose-response curve increased. For soil and sediment samples, the ratio of BIO/CHE ranged from 2.0 to 7.5 among high concentration samples. The ratio of BIO/CHE ranged from 37.3 to 42.3 among low concentration samples. For flue gas samples, when the relative luciferase activity (RLU) was greater than 20000, the result of DRE-Luciferase assay was very close to that of HRGC/HRMS. Based on the preliminary results, the Huh7-DRE-Luc cell is suggested to be qualified for analysis of high concentration samples.