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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 何國榮(Guor-Rong Her) | |
| dc.contributor.author | Chia-Ching Ding | en |
| dc.contributor.author | 丁佳慶 | zh_TW |
| dc.date.accessioned | 2021-06-15T04:49:00Z | - |
| dc.date.available | 2010-08-04 | |
| dc.date.copyright | 2010-08-04 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-03 | |
| dc.identifier.citation | 參考文獻
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45925 | - |
| dc.description.abstract | 核糖核酸酶B醣鏈結構含有M5G2、M6G2、M7G2、M8G2、M9G2(M,Man; G,GlcNAc)五種，其中M7G2具有三種結構異構物。進行醣鏈與醣鏈異構物之結構分析時，需要先利用層析方法將其分離。實驗中，分別以對胺基苯甲酸乙酯(ABEE)和對胺基苯甲酸(ABA)標記高甘露醣鏈，嘗試利用毛細管區帶電泳系統於非揮發性硼酸鹽緩衝溶液分離不同大小的醣鏈與醣鏈異構物。於硼酸鹽緩衝溶液中，對胺基苯甲酸(ABA)標記高甘露醣鏈具有較佳分離效果，但仍無法達到高甘露醣鏈達基線分離與M7G2三種同分異構物之分離。利用添加負電荷環糊精於硼酸緩衝溶液系統，雖然M7G2三種同分異構物仍未達到分離，不同大小的高甘露醣鏈可達基線分離。環糊精和硼酸鹽緩衝溶液對於負離子電噴灑會有嚴重訊號抑制以及電噴灑時會因結晶而造成噴頭阻塞與噴灑不穩定。因此本研究使用實驗室所開發之低鞘流-液體接合式介面以克服上述問題。此介面包含三大部分：液體接合槽、銜接管及低鞘流介面。實驗中將銜接管以塗佈的方式降低電滲流，使得硼酸根離子和環糊精會停留於液體接合槽中，以避免硼酸離子與環糊精進入質譜造成分析物訊號抑制，但仍保有分離效果。 | zh_TW |
| dc.description.abstract | Ribonulease B contains five high-mannose oligosaccharide which are M5G2, M6G2, M7G2, M8G2 and M9G2 (M, Man; G, GlcNAc). Among these glycan, the M7G2 consists of three structural isomers. The separation of these glycan isomers is necessary to characterize its structure. CE was chosen as separation platform because of its high separation efficiency. In our experiment, high-mannose glycans were labeled with p-aminobenzoic acid ethyl ester (ABEE) and aminobenzoic acid (ABA) resepectively, then these derivatives were analyzed by CE under nonvolatile borate buffer system. Our results indicated that separation efficiency was better for ABA-labeled glycans. The high-mannose glycans were partially separated, whereas three isomers of M7G2 coeluted under this condition. To achieve better separation efficiency, sulfated-β-cyclodextrin was added into the borate buffer system. The results show that the high-mannose glycans were baseline separated. However, these nonvolatile species can suppress the analyte signals and crystallize to block the sprayer to terminate ESI process. A Liquid-Junction/Low-Flow interface was utilized to overcome the above problems. The interface consists of a liquid junction reversoir, a coated connecting column and a low flow interface. A coated connecting column was employed to reduce EOF. The coating solutions contained 10% polybrene solution can reduce the EOF (18%) in comparison with non-modified capillary. Under such condition, the borate and cyclodextrin can retain in the liquid-junction revervoir, whereas the analyte can migrate toward the ESI source due to EOF. With this approach, the signals of analyte were not suppressed and the separation efficiency was maintained. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T04:49:00Z (GMT). No. of bitstreams: 1 ntu-99-R97223181-1.pdf: 4004244 bytes, checksum: 14eacd5cbafb3f87aa3a37c1ab1c9d40 (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 第一章 序論
1-1 前言.....................01 1-2 毛細電泳分析核醣核酸酶B中高甘露醣鏈 .........02 1-3 衍生化.....................03 1-4 毛細管電泳...................04 1-5 電噴灑游離法(electrospray ionization ESI) .........11 1-5-1 電灑法原理.................11 1-5-2 負離子電灑法.................12 1-6 非揮發性緩衝溶液.................14 1-6-1 硼酸(borate).................14 1-6-2 環糊精(cyclodextrin) ...............16 1-7 毛細管電泳/電灑法質譜儀介面..............19 1-8 二維離子阱質譜儀.................23 1-9 研究動機...................25 第二章 實驗部份 2-1 藥品、材料....................35 2-2 實驗裝置...................37 2-2-1 毛細管電泳裝置.................37 2-2-2 紫外光偵測器.................37 2-2-3 質譜儀...................37 2-2-4 毛細管塗佈製作流程...............38 2-2-5 毛細電泳質譜介面製作...............39 2-2-5-1 低流速鞘流電灑基座製作...........39 2-2-5-2低流速鞘流-液體結合式介面製作.........40 2-2-6高速過濾篩離法 (Ultrafiltration)............ 41 2-2-7 石墨固相萃取管柱...............41 2-2-8 聚合物固相萃取管柱...............42 2-2-9 吹氣濃縮儀.................42 2-2-10 加熱恆溫反應器................42 2-3 實驗方法...................43 2-3-1 前處理...................43 2-3-1-1醣蛋白醣鏈之切醣及純化........... 43 2-3-1-2對胺基苯甲酸（ABA）開環標記及衍生化醣鏈純化... 44 2-3-1-3 毛細電泳預平衡.............45 2-3-2 毛細電泳紫外光檢測...............45 2-3-3 質譜條件設定.................46 第三章 結果與討論 3-1 對胺基苯甲酸乙酯標記高甘露醣鏈.............54 3-1-1 對胺基苯甲酸乙酯閉環標記高甘露醣鏈.........55 3-1-2 對胺基苯甲酸乙酯開環標記高甘露醣鏈.........57 3-2 對胺基苯甲酸開環標記高甘露醣鏈.............59 3-2-1 對胺基苯甲酸開環標記高甘露醣鏈樣品製備與結構判定.....59 3-2-2 對胺基苯甲酸開環標記高甘露醣鏈之毛細電泳分離條件探討...61 3-2-3 硼酸鹽與S-β-環糊精於對胺基苯甲酸開環標記高甘露醣鏈訊號制..................63 3-2-4 以速鞘流-液體接合式介面減少硼酸鹽與S-β-環糊精之影響.......64 3-2-4-1 銜接管條件探討.............64 3-3-4-2以低流速鞘流-液體接合式介面銜接毛細管分析高甘露醣.......67 3-3 結論.....................69 | |
| dc.language.iso | zh-TW | |
| dc.title | 高甘露醣鏈於非揮發性環糊精和硼酸緩衝溶液之毛細電泳質譜研究 | zh_TW |
| dc.title | CE-MS Analysis of High-Mannose Glycans by Using Nonvolatile Sulfated β-Cyclodextrin and Borate Buffer | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 傅明仁(Ming-Ren Fuh),張耀仁(Yan-Zin Chang) | |
| dc.subject.keyword | 核醣核酸酶,B,高甘露醣鏈,硼酸,環糊精,毛細管塗佈,低鞘流-液體接合式介面, | zh_TW |
| dc.subject.keyword | ribonuclease B,high-mannose glycan,borate,cyclodextrin,Capillary coating,low flow-liquid junction interface, | en |
| dc.relation.page | 100 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-08-03 | |
| dc.contributor.author-college | 理學院 | zh_TW |
| dc.contributor.author-dept | 化學研究所 | zh_TW |
| 顯示於系所單位： | 化學系 | |
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