中山醫學大學機構典藏 CSMUIR:Item 310902500/2710
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    題名: 食品中赭麴毒素之檢測分析及其致毒機制之探討
    Analysis of Ochratoxins in Food and Studies on Its Toxicological Mechanism
    作者: 余豐益
    Yu, Feng-Yih
    貢獻者: 中山醫學院生命科學系
    關鍵詞: 赭麴毒素;酵素免疫分析法;高效液相層析法
    日期: 2005
    上傳時間: 2010-11-05T10:48:57Z (UTC)
    摘要: 赭麴毒素 A (Ochratoxin A, OTA)的多株抗體由赭麴毒素 A 與載體蛋白質接合物 (OTA- γ globulin),免疫到兔子體內，接著收集純化免疫血清來得到辨識赭麴毒素的抗體，直接與非直接競爭型酵素免疫分析法 (Competitive direct and indirect enzyme-linked imunosorbent assay)被用來測定抗體的特性和分析赭麴毒素在遭受污染食品中的含量。此一直接競爭型酵素免疫分析法對赭麴毒素 A 與其類似物赭麴毒素 B 和 C 有弱交叉反應 ,50%抑制濃度分別為 0.9, 110 和 0.54ng/mL。將黃豆樣品加入 10 to 250 ng/g 不同濃度之素以 50%甲醇水溶液萃取後以直接競爭型酵素免疫分析法分析回收率可達 85.9%。目前利用此一酵素免疫分析法分析 20 種不同的農產品中，其中有 12 個樣品遭受到 16 到 160 ng/mL 之污染。樣品中的赭麴毒素亦經由高效液相層析法 (High performance liquid chromatography)來加以分析確認。
    Polyclonal antibodies for ochratoxin A (OTA)were generated from rabbits after the animals had been immunized with either OTA-.gamma.-globulin or OTA-keyhole limpet hemocyanin (KLH). A competitive direct enzyme-linked immunosorbent assay (cdELISA) and a competitive indirect ELISA (ciELISA) were used for the characterization of the antibodies and for analysis of OTA in various agricultural commodities. The antibody titers in the serum of rabbits immunized with OTA-.gamma.-globulin were considerably higher than those in rabbits immunized with OTA-KLH. The antibodies from the rabbits immunized with OTA-.gamma.-globulin were further characterized. In the cdELISA, the concentrations causing 50% inhibition (IC50) of binding of OTA-horseradish peroxidase to the antibodies by OTA, ochratoxin B (OTB), and ochratoxin C (OTC) were found to be 0.90, 110 and 0.54 ng/mL, respectively. When 10 to 250 ng/g of standard OTA was spiked to soybean samples and then extracted with 50% aqueous methanol, the recovery rate of OTA was found to be 85.9% in the cdELISA . Analysis of OTA in various agricultural commodities showed that 12 of the 20 examined samples were contaminated with OTA at levels from 16 to 160 ng/g. The efficacy of cdELISA was also confirmed by the high-performance liquid.
    URI: https://ir.csmu.edu.tw:8080/handle/310902500/2710
    顯示於類別:[生物醫學科學學系暨碩士班] 研究計劃

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