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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李允中 教授 | |
| dc.contributor.author | Jong-Jyh chen | en |
| dc.contributor.author | 陳忠智 | zh_TW |
| dc.date.accessioned | 2021-06-13T04:11:46Z | - |
| dc.date.available | 2006-07-27 | |
| dc.date.copyright | 2006-07-27 | |
| dc.date.issued | 2006 | |
| dc.date.submitted | 2006-07-25 | |
| dc.identifier.citation | 參 考 文 獻
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32572 | - |
| dc.description.abstract | 本研究旨將一般生化分析轉用成具動態且可微量分析之毛細管電泳技術，並進行以下幾種管內反應之應用。首先，為將幾丁質類物質之親合特性作具體化的描述，採親合性電泳技術作幾丁質類物質之結合常數的驗證，本研究係藉由馬尿酸與幾丁寡醣(5〜9mer.)間具親合特性，故分析其親合物之結合常數約為0.56〜1.0174 M-1。由於該分析物在毛細管中的分離與結合係同時發生，所以對於分子間的動態交互反應提供相當多有用的資訊。
另外利用在毛細管管內進行呈色反應，可提高毛細管電泳圖譜分析的靈敏度，本研究以鄰苯二甲醛(O-phthalaldehyde；OPA)及硼酸液與葡萄糖胺可形成複合物，並對該兩種方法作一比較，結果顯示該OPA法較為靈敏(0.1~50mM, A340)且專一，且為克服OPA試液之衰減快速及再現性差的缺點，本研究之OPA法配合毛細管可線上反應、偵測之特點，成功地對於市售含葡萄糖胺之保健產品作有效的量測。 又毛細管電泳管內微萃取技術是另種對於樣品前處理的有效手段，在富含各種蛋白質的市售醬油中，毛細管電泳技術是無法有效地偵測出防腐劑成份的，本研究則將醬油先經酸化、再以乙酸乙酯萃取後之有機層液，微量注入具鹼性緩衝液之毛細管中進行電泳程序，則可簡化對醬油中防腐劑之分析，並達到定量的目的。 | zh_TW |
| dc.description.abstract | Dynamic micro-scaled biochemical processes were designed and conducted within the separation capillary of capillary electrophoresis (CE) system, and several possible applications were demonstrated. To characterize the bio-affinity properties of chitosan, ACE (affinity capillary electrophoresis) techniques were applied to evaluate the binding constant. From the shift in migration times of hippurate / chitooligosacharide complex, the binding constant of hippurate with chitooligosacharides (around 5〜9 mer.) was calculated to be in the range of 0.56〜1.0174 M-1. Since the binding process occurred simultaneously with the separation process, the method is rapid and provides valuable information about the dynamic molecular interactions.
Secondly, in-capillary chromogenic reaction was used to enhance the separation and sensitivity of CE system. Two labeling procedures for glucosamine using o-phthalaldehyde (OPA) and borate were developed and compared. The OPA method is more sensitive (0.1~50mM, A340) and specific, but the reaction is too quick to reproducibly operate in an off-line manner. OPA-labeling reaction was successfully performed on-line within the capillary, and the glucosamine contents in nutraceuticals were accurately measured. Thirdly, in-capillary micro-extraction process served as an efficient sample pretreatment procedure. Benzoate derivatives are common preservatives in soy sauce; however, the CEgrams of untreated samples were too complicated and can not be effectively resolved / analyzed. The complicated samples were acidified and then extracted with ethyl acetate. The organic extract was directly injected and analyzed in capillary zone electrophoresis mode. The organic sample plug (benzoates in ethyl acetate) was extracted and then separated in the alkaline running buffer, which simplified the electropherogram for quantification. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T04:11:46Z (GMT). No. of bitstreams: 1 ntu-95-D89631003-1.pdf: 893049 bytes, checksum: f6104bf1a4e2a2661f70c4c0845babd3 (MD5) Previous issue date: 2006 | en |
| dc.description.tableofcontents | 誌謝 i
中文摘要……………………………………………………………… ii 英文摘要……………………………………………………………… iv 目錄…………………………………………………………………… vi 圖目錄………………………………………………………………… x 表目錄………………………………………………………………… xii 符號說明……………………………………………………………… xiii 字詞縮寫……………………………………………………………… xvi 第一章 前言………………………………………………………… 1 第二章 文獻探討…………………………………………………… 3 2-1 毛細管電泳…………………………………………………… 5 2-1-1 毛細管之性質…………………………………………… 6 2-1-2 緩衝液之作用…………………………………………… 7 2-1-3 電滲泳在分析上之影響………………………………… 8 2-1-4 毛細管電泳檢測指標…………………………………… 9 2-2 改善毛細管電泳之方法……………………………………… 11 2-2-1 面積的正規化…………………………………………… 11 2-2-2 樣品注入的機制及其特性……………………………… 12 2-2-3 溫度對毛細管電泳之影響……………………………… 15 2-3 親合性毛細管電泳之定義及分類…………………………… 17 2-3-1 親合性電泳的特點……………………………………… 21 2-3-2 運用親合性電泳解結合常數之注意事項……………… 24 2-3-3 運用親合性電泳的時機………………………………… 26 2-3-4 運用親合性毛細管電泳分析醣類分子………………… 27 2-3-5 結合常數(KB)之運用…………………………………… 28 2-4 幾丁質類物質………………………………………………… 32 2-4-1 幾丁質類物質之去乙醯度……………………………… 34 2-4-2 幾丁質類物質之降解…………………………………… 35 2-4-3 葡萄糖胺之特性………………………………………… 37 2-4-4 葡萄糖胺之檢測………………………………………… 38 2-5 萃取之機制…………………………………………………… 40 2-5-1 萃取技術之應用………………………………………… 41 2-5-2 苯甲酸及其衍生物……………………………………… 42 第三章 試驗設備與方法…………………………………………… 44 3-1 藥劑…………………………………………………………… 44 3-1-1 幾丁質類物質之分析試驗……………………………… 44 3-1-2 馬尿酸鈉與幾丁寡醣之親合性電泳…………………… 45 3-1-3 防腐劑之分析試驗……………………………………… 45 3-1-4 酸水解程序……………………………………………… 46 3-1-5 鄰苯二甲酸(OPA)溶液之製備…………………………… 46 3-2 儀器設備與方法……………………………………………… 47 3-2-1 儀器……………………………………………………… 47 3-2-2 馬尿酸鈉與幾丁寡醣之親合性電泳…………………… 47 3-2-3 幾丁質類物質酸水解物試驗…………………………… 48 3-2-4 硼酸鹽與葡萄醣胺之試驗……………………………… 48 3-2-5 鄰苯二甲酸與葡萄醣胺之試驗………………………… 48 3-2-6 防腐劑之分析…………………………………………… 49 3-2-7 回收率試驗……………………………………………… 49 第四章 結果與討論………………………………………………… 50 4-1 幾丁寡醣之毛細管電泳……………………………………… 50 4-1-1 馬尿酸鈉與幾丁寡醣之親合性電泳…………………… 50 4-1-2 馬尿酸鈉與幾丁寡醣之結合常數……………………… 54 4-2 幾丁質類物質之酸水解……………………………………… 59 4-2-1 乙酸、葡萄糖胺、N-乙醯葡萄糖胺之分析……………… 59 4-2-2 去乙醯度之分析………………………………………… 61 4-3 葡萄糖胺(Glucosamine)之檢測……………………………… 68 4-3-1 硼酸鹽與葡萄醣胺之反應……………………………… 68 4-3-2 N-乙醯葡萄糖胺與葡萄糖胺之分析…………………… 70 4-3-3 硼酸鹽-葡萄醣胺衍生物檢量線之建立……………… 72 4-3-4 市售樣品測試及分析（硼酸鹽法）……………………… 74 4-3-5 OPA試劑與葡萄醣胺之反應……………………………… 76 4-3-6 硼酸鹽緩衝液濃度之影響……………………………… 78 4-3-7 工作電壓之影響………………………………………… 80 4-3-8 硼酸緩衝液pH之影響…………………………………… 82 4-3-9 OPA與葡萄醣胺衍生物之最適混合時間分析………… 84 4-3-10 OPA與葡萄醣胺衍生物檢量線之建立………………… 88 4-3-11 市售樣品測試及分析（OPA法） ………………………… 91 4-3-12 硼酸鹽法與OPA法對葡萄醣胺產品之比較…………… 94 4-4 苯甲酸及其衍生物之分析…………………………………… 96 4-4-1 硼酸緩衝液之電壓-電流(V-I)對應曲線……………… 96 4-4-2 防腐劑之毛細管電泳分離條件………………………… 102 4-4-3 各防腐劑之標準迴歸曲線……………………………… 107 4-4-4 市售含防腐劑之醬油分離……………………………… 107 4-4-5 防腐劑之回收率………………………………………… 111 第五章 結論………………………………………………………… 113 參考文獻……………………………………………………………… 115 附錄 計算結合常數( )之Scatchard Plot模式推導…………… 129 圖目錄 圖2-1 幾丁質類物質之結構式…………………………………… 32 圖4-1 添加幾丁寡醣對馬尿酸鈉之親合性反應………………… 52 圖4-2 幾丁寡醣/馬尿酸鈉之散佈圖(Scatchart plot)…………… 58 圖4-3 施加輔助壓對乙酸之電泳影響…………………………… 60 圖4-4 幾丁質(Chitin)之酸水解物電泳分析…………………… 62 圖4-5 乙酸/葡萄糖胺之分子量比標準曲線…………………… 65 圖4-6 硼酸鹽與葡萄糖胺之結合反應變化情形………………… 69 圖4-7 葡萄糖胺、乙醯葡萄糖胺與硼酸鹽反應後之電泳分析… 69 圖4-8 硼酸鹽/葡萄糖胺濃度比檢量線（馬尿酸為內標準）…… 74 圖4-9 市售含葡萄糖胺錠劑之電泳分析 (硼酸鹽法) …………… 75 圖4-10 鄰苯二甲酸/葡萄糖胺（OPA/GluN）衍生化之吸光反應… 77 圖4-11 硼酸緩衝液濃度對OPA/葡萄糖胺之電泳影響…………… 79 圖4-12 工作電壓對OPA/葡萄糖胺之電泳影響…………………… 81 圖4-13 硼酸緩衝液pH對OPA/葡萄糖胺之電泳影響……………… 83 圖4-14 OPA/葡萄糖胺/OPA夾注模式之電泳圖…………………… 85 圖4-15 OPA/葡萄糖胺/OPA之理論板數及解析度圖……………… 86 圖4-16 OPA/葡萄糖胺衍生物濃度檢量線…………………………… 89 圖4-17 OPA/葡萄糖胺衍生物濃度檢量線（Gycine為內標準）……… 90 圖4-18 市售含葡萄糖胺錠劑之電泳分析………………………… 92 圖4-19 市售含葡萄糖胺錠劑與標準品之頻譜比較………………… 93 圖4-20 毛細管電泳之電壓-電流-電阻之變化………………… 98 圖4-21 數值分析毛細管電泳之適切工作電壓………………… 99 圖4-22 工作電壓對防腐劑分離效果之影響…………………… 103 圖4-23 硼酸緩衝液之pH 值對電泳圖之影響………………… 106 圖4-24 市售醬油直接注入毛細管進行電泳之頻譜圖………… 109 圖4-25 醬油經毛細管電泳線上萃取法所得到之頻譜圖……… 110 表目錄 表4-1 幾丁寡醣添加量對於泳動時間之影響……………………… 53 表4-2 泳動時間之正規化處理……………………………………… 56 表4-3 幾丁質類物質去乙醯度之分析……………………………… 67 表4-4 市售含葡萄糖胺產品之CE比較（OPA法及硼酸鹽法）…… 95 表4-5 工作電壓變異容許程度之評估……………………………… 100 表4-6 各防腐劑相對於馬尿酸鈉之含量迴歸……………………… 108 表4-7 添加於醬油中之防腐劑回收率……………………………… 112 | |
| dc.language.iso | zh-TW | |
| dc.subject | 毛細管電泳 | zh_TW |
| dc.subject | 微萃取 | zh_TW |
| dc.subject | 保健食品 | zh_TW |
| dc.subject | 葡萄糖胺 | zh_TW |
| dc.subject | Capillary electrophoresis | en |
| dc.subject | Glucosamine | en |
| dc.subject | Micro-extraction | en |
| dc.subject | Nutraceutical | en |
| dc.title | 毛細管電泳管內反應之探討和應用 | zh_TW |
| dc.title | On In-capillary Processes of Capillary Electrophoresis and the Applications | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 94-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 陳榮輝 教授,盧福明 教授,楊景雍 助理教授,陳力騏 副教授 | |
| dc.subject.keyword | 毛細管電泳,葡萄糖胺,保健食品,微萃取, | zh_TW |
| dc.subject.keyword | Capillary electrophoresis,Glucosamine,Nutraceutical,Micro-extraction, | en |
| dc.relation.page | 134 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2006-07-26 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物產業機電工程學研究所 | zh_TW |
| 顯示於系所單位： | 生物機電工程學系 | |
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