臺灣博碩士論文加值系統

類黃酮及其配醣體具有多項之生理活性，普遍存在於植物中。目前雖然已有許多關於類黃酮配醣體結構、活性之相關文獻，其中亦有許多分析鑑定之方法。但是結合高效鑑定及活性篩選裝置用以直接分析類黃酮配醣體之系統，仍尚有許多問題待解決。例如，其中以同步檢測抗氧化能力之原理進行之方法而言，一般為利用抗氧化劑其具有清除自由基 [(DPPH, 2,2`-diphenyl-1-picrylhydrazyl) 或 (ABTS, (2,2`-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)] 等之能力。但含有配醣體結構之物質，與 DPPH 或 ABTS 反應效果並不好，所以實驗結果會失真。對此最近有研究團隊提出以電化學分析法取代自由基清除法，偵測含有配醣體之類黃酮，但可惜的是其施加之電壓須大於 1.2V，才能測得類黃酮配醣體。而由於高電壓會使許多原來不屬於抗氧化劑之物質氧化造成嚴重之基質干擾，進而影響到偵測結果。故本研究擬發展一套低電壓之電化學偵測法，結合液相層析及並聯電噴灑質譜儀進行複雜成分之同步抗氧化活性測定與分析物之結構鑑定。本次試驗以五種類黃酮配醣體做為模式化合物，測試其在本電化學偵測上之可行性。而在實驗結果中顯示，本研究系統可以在不同 pH 值之環境下，僅以施加 0.3V 之電壓便能有效偵測到分析物。最後，針對 quercetin-3,4`-di-O-glucopyranoside 與 DPPH 或 ABTS 反應訊號相比較，在電化學分析系統下其偵測靈敏度比 ABTS 分析系統強 2 倍以上，而在 DPPH 之分析系統下則是無法偵測到quercetin-3,4`-di-O-glucopyranoside。

Flavonoid glycosides of plants have multiple physiological activities. A lot of researchers have studied antioxidant activities and structures of flavonoids. Meanwhile, varieties of analytical methods were developed to meet the need of the activity and structure determination. Among which hyphenated devices for simultaneous antioxidant assay and structure identification are currently most promising for high throughput analyses. However, the application of the hyphenated devices in activity assay of flavonoid glycosides is still hindered by many problems. One of the disadvantages is the free radicals [DPPH, 2,2`-diphenyl-1-picrylhydrazyl or ABTS, 2,2`-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)] reacting slowly toward the glycosides. Although there is an alternative electrochemical detecting method attempted to substitute the free radical scavenging methods for flavonoid glycosides detection, the applied voltage must greater than 1.2V, which in fact will cause analytes other than flavonoid glycosides to oxidize. In the present study, we developed a low-voltage driven electrochemical detection technique coupled with HPLC and ESI-MS to inquired antioxidant capacities and structure identification of flavonoid glycosides simultaneously in complicated samples. The device was proved feasible by five model flavonoid glycosides. For effectively detecting the analytes, the applied voltage can downward even to 0.3V and performed consistently under mobile phases of different pH values. Finally, the relative sensitivity of the electrochemical detection to ABTS assay system for quercetin-3,4`-di-O-glucopyranoside is two times, while there was no response in DPPH assay system.

目錄﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍Ⅰ
圖次﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍Ⅱ
表次﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍Ⅵ
中文摘要﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍Ⅶ
英文摘要﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍Ⅸ
壹、前言﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍1
貳、文獻整理﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍4
一、類黃酮及其配醣體之簡介﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍4
二、抗氧化劑作用原理﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍6
三、類黃酮及其配醣體檢測方法﹍﹍﹍﹍﹍﹍﹍﹍﹍8
叁、材料與方法﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 29
一、藥品與材料﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 29
二、儀器設備﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 32
三、溶液配製﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 33
肆、實驗方法﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 35
一、系統之建立﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 35
二、HPLC 分析條件﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 35
三、靜相與流動相之電化學偵測﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 38
四、質譜分析條件﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 45
伍、結果與討論﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 46
一、Model Compounds 之選擇﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 46
二、靜相電化學偵側﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 46
三、流動相電化學偵測之最佳條件探討﹍﹍﹍﹍﹍﹍﹍﹍﹍ 50
四、定性分析﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 66
陸、結論﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 80
柒、參考文獻﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 81

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