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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40133完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李水盛,郭錦樺 | |
| dc.contributor.author | Chia-Wen Lee | en |
| dc.contributor.author | 李嘉雯 | zh_TW |
| dc.date.accessioned | 2021-06-14T16:41:38Z | - |
| dc.date.available | 2009-09-11 | |
| dc.date.copyright | 2008-09-11 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-31 | |
| dc.identifier.citation | 第一部分:
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40133 | - |
| dc.description.abstract | 中文摘要
第一部分：以毛細管電泳分析赤楊屬植物及生物樣品中之oregonin成分 Oregonin為一種廣泛分布於赤屬植物中的diarylheptanoid glycoside，許多研究顯示其具有顯著的抗發炎、抗氧化、抗癌等活性。本研究首次使用毛細管電泳方法之毛細區帶電泳模式（capillary zone electrophoresis），分析生物樣品以及多種赤楊屬植物萃出物中的oregonin成分，分析方法開發過程中分別針對緩衝溶液濃度、緩衝溶液酸鹼值以及施加電壓等參數，進行分析條件的最適化，最後選取pH 8.0，濃度為30 mM之四硼酸鈉溶液作為電泳液，施加電壓與偵測波長分別為30 kV與220 nm。於最適化分析條件下，oregonin能在6分鐘之內完成分析，本分析方法遷移時間之次與次間重複性與中間精密度之RSD小於1.36 %，oregonin與內標準物峰面積比值之次與次間重複性與中間精密度RSD小於1.55 %。本研究並將此條件應用於分析台灣赤楊葉部萃出物、多種赤楊屬植物種子萃出物以及生物樣品中之oregonin，關於oregonin 的安定性亦在研究中加以探討，上述結果證明所開發的分析條件可應用於天然物的研究工作。 第二部分：以UPLC-MS/MS定量在馬兜鈴及生物樣品中之aristolochic acid I及aristolochic acid II 馬兜鈴酸主要存在於馬兜鈴科植物中，其具有強腎毒性及致腫瘤毒性。馬兜鈴酸由一群結構類似的化合物組成，其中又以aristolochic acid I （AAI）以及aristolochic acid II (AAII)被指出與腎毒性具有最大的關聯。本研究首次採用UPLC-MS/MS建立AAI及AAII的分析方法，並用以定量馬兜鈴中及生物樣品中的AAI與AAII含量。最適化分析條件使用pH 3.0, 10 mM甲酸銨緩衝溶液及乙腈作為液相層析的移動相，AA I與AA II可於10分鐘內達基線分離。研究中並針對質譜儀參數作最適化的調整，其中包括：樣品錐電壓、毛細管高電壓、撞擊能量、離子源溫度、去溶劑氣體溫度、去溶劑氣體流速及離子源與樣品錐間距等，定量方法使用選擇反應監測模式（Selected reaction monitoring，SRM）進行AAI及AAII的分析。本分析方法遷移時間之次與次間重複性與中間精密度之RSD小於1.25 %，AAI及AAII峰面積之次與次間重複性與中間精密度RSD小於5.74 %，準確度介於114.51至98.11 %之間，並對於AA I及AA II分別提供了0.14 ng /ml及0.26 ng /ml的偵測極限。本研究使用UPLC-MS/MS建立一個快速而靈敏的AA I與AA II分析方法。 | zh_TW |
| dc.description.abstract | Abstract
Part I: Determination of oregonin in Alnus plants and biological samples by capillary electrophoresis Oregonin, existing primarily in the Alnus plants, displayed anti-inflammatory and antioxidative activities. The capillary zone electrophoresis (CZE) method was developed in this study to quantitatively determine oregonin content in the Alnus plants and biological fluid for the first time. Various parameters, including buffer concentration, pH and applied voltage, were evaluated for their optimum analytical conditions. The optimized buffer was composed of 30 mM sodium tetraborate at pH 8.0. The separation voltage was set at 30 kV and the UV detection wavelength was set at 220 nm. Oregonin could be determined within 6 minutes under such optimized conditions. Relative standard deviation (RSD) of the run-to-run repeatability and intermediate precision of the retention time of oregonin was within 1.36 %. Run-to-run repeatability and intermediate precision of the peak area ratios of oregonin to internal standard, theophylline, were both within 1.55 % RSD. The presented method was applied to analyze oregonin in leaves of of Alnus formosana, seeds of various Alnus plants as well as biological samples. The stability of oregonin in biological system was indicated in this study. It demonstrates the potential of this developed method in natural product research. Part II: Determination of aristolochic acid I and aristolochic acid II in Aristolochia debilis and biological samples by UPLC-MS/MS Aristolochic acids (AAs), existing primarily in the Aristolochia plants, shown potent nephrotoxicity and carcinogenicity. AAs are a mixture of structure related compounds, where aristolochic acid I (AA I), and aristolochic acid II (AA II) are reported to be correlated with Chinese herbs nephropathy (CHN). An UPLC-MS/MS method was developed in this study to quantitatively determination of aristolochic acid I, and aristolochic acid II in Aristolochia debilis and biological fluid for the first time. By using pH3.0, 10mM ammonium formate buffer and acetonitrile as mobile phase, Aristolochic acids could be determined within 10 minutes. The mass parameters including cone voltage, capillary voltage, collision energy, ion source temperature, desolvation temperature, desolvation gas flow and the probe distance were subsequently optimized. Selected reaction monitoring (SRM) method was used for quantitative analysis. Relative standard deviation (R.S.D.) of the run-to-run repeatability and intermediate precision of the retention time of AAI and AAII was within 1.25%. Run-to-run repeatability and intermediate precision of the peak area of AAI and AAII were both within 5.74% R.S.D. Accuracy of the method was between 98.11% and 114.51%. Limit of detections was 0.14 ng/ml for AAI and 0.26 ng/ml for AAII. The result shows that the present UPLC-MS/MS method is efficient, accurate and sensitive for the determination of AAI and AAII. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-14T16:41:38Z (GMT). No. of bitstreams: 1 ntu-97-R95423007-1.pdf: 1704987 bytes, checksum: 6ea20cd8a3c39b199b1d99489ab81559 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 口試委員審定書…………………………………………………………………………i
誌謝……………………………………………………………………………………ii 中文摘要………………………………………………………………………………iii 英文摘要…………………………………………………………………………………v 目錄……………………………………………………………………………………viii 圖目錄…………………………………………………………………………………xiii 表目錄…………………………………………………………………………………xvii 論文研究主旨…………………………………………………………………………1 第一部分 以毛細管電泳分析赤楊屬植物及生物樣品中之oregonin成分 壹、序論………………………………………………………………………………2 11 毛細管電泳原理及其發展概要………………………………………………2 111電泳發展概要…………………………………………………………2 112毛細管電泳原理概述2 12 本論文所使用之毛細管電泳方法……………………………………………5 貳、研究動機…………………………………………………………………………5 21 oregonin活性相關研究……………………………………………………6 22 應用毛細管電泳於oregonin之分析方法開發…… ………………………7 參、實驗部分…………………………………………………………………………8 31 儀器…………………………………………………………………………8 32 藥品與試劑…………………………………………………………………8 33 標準品貯液之製備…………………………………………………………8 34 植物萃取……………………………………………………………………9 35 動物試驗與樣本收集…………………………………………………………9 36 檢品溶液之製備……………………………………………………………9 361植物萃出物之檢品溶液……………………………………………9 362血清樣品前處理………………………………………………………10 363 動物試驗樣品檢品溶液……………………………………………10 37毛細管電泳系統……………………………………………………………10 38毛細管之處理…………………………………………………………………11 39分析方法之確效…………………………………………………………12 391 精確度 (precision)…………………………………………………12 392 線性 (linearity)……………………………………………………12 393 靈敏度 (sensitivity)………………………………………………13 394 準確度 (accuracy)…………………………………………………13 3941 植物萃取物………………………………………………13 3942 生物樣品…………………………………………………13 肆、結果與討論………………………………………………………………………14 41分析方法之建立………………………………………………………………14 411緩衝溶液酸鹼值………………………………………………………14 412緩衝溶液之濃度………………………………………………………15 42最佳分析條件…………………………………………………………………15 43分析方法之確效………………………………………………………………15 431精確度 (precision)…………………………………………………15 4311一日之間重複性 (repeatability, intra-day precision)………15 4312中間精密度（Intermediate precision, inter-day precision）……15 432線性 (linearity) ………………… …………………………………16 433靈敏度 (sensitivity)……………………………………………………16 434 準確度 (accuracy)…………………………………………………16 44分析方法之應用……………………………………………………………17 441 赤楊屬植物oregonin成分之分析與定量……………………………17 442 生物性樣品之分析…………………………………………………17 伍、結論……………………………………………………………………………19 陸、參考文獻…………………………………………………………………………20 第二部分：以UPLC-MS/MS定量在馬兜鈴及生物樣品中之Aristolochic acid 與Aristolochic acid II 壹、 序論………………………………………………………………………………45 11 LC-MS簡介…………………………………………………………………45 111 電灑游離法（Electrospray ionization，ESI）………………………46 112 大氣壓化學游離法（atmospheric pressure chemical ionization，APCI） ……………………………………………………………………47 12 超效液相層析（Ultra performance liquid chromatography，UPLC） ………………………………………………………………………………47 121 液相層析法之分離效率……………………………………………47 122超效液相層析（Ultra performance liquid chromatography，UPLC） ………………………………………………………………………47 貳、 研究動機…………………………………………………………………………49 21植物簡介……………………………………………………………………51 22 馬兜鈴酸 ……………………………………………………………………52 221馬兜鈴酸及其衍生物 ………………………………………………52 222馬兜鈴酸引起之中藥腎病變 ………………………………………52 23 補肺阿膠湯 …………………………………………………………………53 24 文獻使用之分析方法………………………………………………………54 25 以UPLC-MS/MS進行馬兜鈴及生物樣品中之Aristolochic acid I與Aristolochic acid II定量分析 ………………………………………………56 參、實驗部分…………………………………………………………………………58 31 儀器…………………………………………………………………………58 32 藥品與試劑…………………………………………………………………58 33 標準品貯液之製備…………………………………………………………59 34藥材萃取……………………………………………………………………59 35 動物試驗與樣本收集………………………………………………………59 36 血清樣品前處理……………………………………………………………59 37 檢品溶液之製備……………………………………………………………60 371 標準品溶液………………………………………………………… 60 372 植物萃出物之檢品溶液……………………………………………60 38 UPLC-MS與UPLC-UV條件………………………………………………61 39 半製備型HPLC 條件………………………………………………………62 310 分析方法之確效…………………………………………………………62 3101 精確度 （precision）………………………………………………62 3102 線性 (linearity)……………………………………………………62 3103 靈敏度 (sensitivity)…………………………………………………62 3104 準確度 (accuracy)…………………………………………………62 肆、結果與討論………………………………………………………………………64 41 分析方法之建立……………………………………………………………64 42 層析條件之選取……………………………………………………………64 421緩衝溶液之選取………………………………………………………65 422 有機相溶劑之選取…………………………………………………65 423 緩衝水溶液之酸鹼值………………………………………………65 424 層析條件使用之流速………………………………………………66 43質譜儀參數之最適 …………………………………………………………66 431 離子源之選取……………………………………………………66 432 樣品錐電壓…………………………………………………………67 433 毛細管高電壓………………………………………………………67 434 撞擊能量……………………………………………………………67 435 去溶劑氣體溫度……………………………………………………67 436 離子源溫度…………………………………………………………68 437 去溶劑氣體流速……………………………………………………68 438 離子源至樣品錐之間距離…………………………………………68 44最佳分析條件………………………………………………………………68 45分析方法之確效……………………………………………………………69 451精確度…………………………………………………………………69 4511一日內重複性………………………………………………………69 4512中間精密度…………………………………………………………70 452線性……………………………………………………………………70 453靈敏度…………………………………………………………………70 454 準確度…………………………………………………………………70 46分析方法之應用…………………………………………………………71 伍、結論………………………………………………………………………………72 陸、參考文獻…………………………………………………………………………73 | |
| dc.language.iso | zh-TW | |
| dc.subject | 選擇反應監測模式 | zh_TW |
| dc.subject | 電噴霧游離 | zh_TW |
| dc.subject | 超效液相層析-三段式四極棒串聯式質譜儀 | zh_TW |
| dc.subject | 中草藥腎病變 | zh_TW |
| dc.subject | 馬兜鈴 | zh_TW |
| dc.subject | 馬兜鈴酸 | zh_TW |
| dc.subject | 生物樣品 | zh_TW |
| dc.subject | 毛細區帶電泳 | zh_TW |
| dc.subject | Oregonin | zh_TW |
| dc.subject | 赤楊屬植物 | zh_TW |
| dc.subject | Alnus plants | en |
| dc.subject | Oregonin | en |
| dc.subject | Capillary zone electrophoresis | en |
| dc.subject | Biological samples | en |
| dc.subject | Aristolochic acid | en |
| dc.subject | Aristolochia debilis | en |
| dc.subject | Chinese herbs nephropathy (CHN) | en |
| dc.subject | UPLC-MS/MS | en |
| dc.subject | ESI | en |
| dc.subject | Selected reaction monitoring (SRM) | en |
| dc.title | 第一部分：以毛細管電泳分析赤楊屬植物及生物樣品中之oregonin成分
第二部分：以UPLC-MS/MS定量在馬兜鈴及生物樣品中之 aristolochic acid I及aristolochic acid II | zh_TW |
| dc.title | Part I: Determination of oregonin in Alnus plants and biological samples
by capillary electrophoresis Part II: Determination of aristolochic acid I and aristolochic acid II in Aristolochia debilis and biological samples by UPLC-MS/MS | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳繼明,林哲輝 | |
| dc.subject.keyword | Oregonin,赤楊屬植物,毛細區帶電泳,生物樣品,馬兜鈴酸,馬兜鈴,中草藥腎病變,超效液相層析-三段式四極棒串聯式質譜儀,電噴霧游離,選擇反應監測模式, | zh_TW |
| dc.subject.keyword | Oregonin,Alnus plants,Capillary zone electrophoresis,Biological samples,Aristolochic acid,Aristolochia debilis,Chinese herbs nephropathy (CHN),UPLC-MS/MS,ESI,Selected reaction monitoring (SRM), | en |
| dc.relation.page | 96 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2008-08-01 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 藥學研究所 | zh_TW |
| 顯示於系所單位： | 藥學系 | |
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