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研究生:林慧雅
論文名稱:利用微透析技術連結高效能液相層析儀檢測銀杏葉萃取液之銀杏內酯A、B和白果內酯
指導教授:邱鴻麟邱鴻麟引用關係
學位類別:碩士
校院名稱:國立高雄師範大學
系所名稱:化學系
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:90
中文關鍵詞:微透析技術銀杏內酯高效能液相層析
相關次數:
  • 被引用被引用:3
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本研究利用微透析樣品前處理裝置連結高效能液相層析儀來檢測銀杏葉萃取液中之白果內酯和銀杏內酯A、B,探討層析管柱及動相條件,找出最佳液相層析條件;探討透析流速和收集時間、灌注液之極性修飾劑濃度、鹽析效應、樣品溶液pH值影響,找出最佳之微透析條件。
最佳層析條件為使用逆相C-18層析管柱,以及甲醇/氰甲烷/1mM、pH=5.0磷酸緩衝溶液(30:5:65),流速1.0 ml/min之沖提液;最佳透析條件為使用100%甲醇為灌注液、流速為2.12μL/min,收集20分鐘。
最佳實驗條件下,分析物的透析回收率分別為:白果內酯104%,銀杏內酯A、B為103%、105%,白果內酯和銀杏內酯A、B之偵測極限分別為6ppm、9ppm、9ppm。
The purpose of the study was to analyze ginkgolide A、B and bilobalide in Ginkgo biloba L. extractions by micro-dialysis connected with high-performance liquid chromatographic (HPLC). The conditions of HPLC analysis included the stationary phase selection and the mobile phase condition were examined. Also the conditions of micro-dialysis which included flow rate, the modifier of perfusate, the concentration of salt adding of the sample as well as the pH were examined.
The optimal conditions of HPLC analysis were C-18 reversed-phase column with the flow rate of 1.0 mL/min of the mobile phase of methanol/acetonitrile/ pH=5、10-3 M phosphate buffer(30:5:65). And the optimal conditions of the micro-dialysis sampling were 100% methanol as perfusate with the flow rate of 2.12 μL/min and 20 min. collection time.
By the optimal conditions, the recoveries of micro-dialysis were 104%, 103%, 105% for bilobalide and ginkgolides A, B, respectively. The detection limites of 6ppm, 9ppm, 9ppm were found for bilobalide and ginkgolides A, B, respectively.
第一章 緒論...............................................................................................1
一、 前言.............................................................................................1
二、 銀杏概述.....................................................................................4
1. 銀杏...........................................................................................4
2. 銀杏葉的藥理性質和功用.......................................................8
3. 銀杏的主要組成和結構.........................................................13
4. 銀杏內酯和白果內酯的偵測方法.........................................19
三、 微透析分析方法概述...............................................................20
(一) 微透析理論之介紹...............................................................20
1. 立體障礙排除.................................................................22
2. 分配.................................................................................22
3. 擴散.................................................................................23
4. Donnan平衡理論...........................................................23
5. 靜電作用效應.................................................................24
(二)透稀薄膜的介紹....................................................................25
(三)微透析技術應用.....................................................................27
(四)微透析的相對回收.................................................................26
(五)微透析技術應用.....................................................................26
四、研究動機.....................................................................................30
第二章 研究方法....................................................................................31
一、 實驗藥品、器材、儀器設備....................................................31
1. 藥品.......................................................................................31
2. 器材.......................................................................................32
3. 儀器設備...............................................................................33
二、 藥品配製...................................................................................35
(一) 沖提液的配製.................................................................35
(二) 儲備標準溶液1000ppm的配製....................................36
(三) 標準溶液的配製.............................................................36
(四) 鹽析溶液的配製.............................................................36
1. 配製5M氯化鈉水溶液...............................................36
2. 混合標準品中添加不同濃度之氯化鈉水溶液.........37
(五) 銀杏葉萃取液的配製.....................................................37
1. 銀杏葉的處理與保存.................................................37
2. 銀杏葉萃取液.............................................................37
三、 實驗過程...................................................................................38
(一)實驗玻璃器皿矽烷化過程..............................................38
(二)透析薄膜的探討.............................................................39
1. 透析裝置的組裝及結構圖.........................................39
2. 透析薄膜使用前之準備及清洗與保存.....................40
3. 中空透析薄膜的使用壽命.........................................41
(三)液相層析方法的建立......................................................41
1. UV偵測波長的選擇...................................................41
2. 層析管柱的選擇.........................................................42
3. 動相的選擇.................................................................42
(四)微透析分析方法的建立...................................................43
1. 灌注液流速和收集時間的選擇...................................43
2. 極性修飾劑濃度的影響...............................................44
3. 鹽類效應的影響...........................................................45
4. 樣品溶液pH值的選擇.................................................45
四、 透析暨層析方法再現性測試...................................................46
五、 校正曲線的建立.......................................................................46
六、 偵測極限的測定.......................................................................47
七、 真實樣品...................................................................................47
第三章 結果與討論.................................................................................48
一、 液相層析方法的建立...............................................................48
1. UV偵測波長的選擇.............................................................48
2. 層析管柱的選擇...................................................................50
3. 動相的選擇...........................................................................50
二、 微透析分析方法的建立..........................................................53
1. 透析流速和收集時間的探討...............................................53
2. 灌注液中極性修飾劑濃度的影響.......................................55
3. 鹽析效應的影響...................................................................58
4. 樣品溶液pH值的影響.........................................................61
三、 透析暨層析方法再現性的測試. .............................................65
四、 校正曲線的建立及其偵測極限...............................................66
五、 透析回收率的探討...................................................................68
六、 真實樣品的分析.......................................................................68
第四章 結論.............................................................................................72
第五章 參考文獻.....................................................................................74
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