臺灣博碩士論文加值系統

在本研究中，我們開發了三種液相層析串聯質譜（Liquid Chromatography/ Mass Spectrometry，LC/MS）方法來辨認藥材中的化學指標成分，分別針對生曬參、紅參及西洋參中的六個人參皂苷（Ginsenosides Rb1、Ginsenosides Rg1、Ginsenosides Rc、Ginsenosides Rg3、Ginsenosides Re和Ginsenosides Rh2），枳殼中的七個黃酮類化合物（Synephrine、Narirutin、Naringin、Hesperidin、Quercertin、Naringenin 和Imperatorin）與黃耆及紅耆中的七個黃酮類化合物（Calycosin-7-O-β-D-glucoside、Calycosin-7-O-β-D-glucoside -6-O-malonate、Ononin、Calycosin、Formononetin-7 -O-β-D -glucoside-6-O-malonate、Astragaloside IV和 Formononetin）進行分析鑑定。
此外，我們還利用LC/MS/MS對黃耆中的二個黃酮類化合物（Astragaloside IV和Formononetin）進行二次質譜的鑑定。
液相層析部分，靜相是使用Cosmosil ARII C18 5μm 2.1×150mm的管柱，移動相是使用醋酸緩衝液及Acetonitrile進行梯度沖提，而偵測波長分別為210nm、254nm和280nm。
串聯質譜部分，離子化方式是以電噴灑法（Electrospray Ionization，ESI），質量分析儀採用四極柱（Quadrupole）和飛行時間（Time of Flight）組合，帶動氣體（Carrier gas）使用氮氣，而碰撞氣體（Collision gas）使用氬氣。
並使用對照成份以進行辨認生曬參、紅參、西洋參、枳殼、黄耆及紅耆，於紫外光光譜與質譜中的化學成分，並製作藥材的化學指紋圖譜，利用這些化學指紋圖譜，可區別這些藥材的基原品種。

Liquid chromatography/mass spectrometry ( LC/MS ) methods were developed to identify six ginsenosides ( Ginsenosides Rb1, Ginsenosides Rg1, Ginsenosides Rc, Ginsenosides Rg3, Ginsenosides Re and Ginsenosides Rh2 ) in Panax ginseng and Panax quinquefolinm , seven flavonoids ( Synephrine, Narirutin, Naringin, Hesperidin, Quercertin, Naringenin and Imperatorin ) in Citrus aurantium and seven flavonoids ( Calycosin-7-O-β-D-glucoside, Calycosin-7-O-β-D-glucoside -6-O-malonate, Ononin, Calycosin, Formononetin-7-O-β-D-glucoside-6-O-malonate, Astragaloside IV and Formononetin ) in Astragalus membranaceus and Hedysarum polybotrys.
LC was carried out using a C18 column with linear gradient elution of acetate buffer- acetonitrile and detected at the ultraviolet (UV) wavelength of 210nm, 254nm and 280nm, separately.
The tandem MS was used with positive-ion electrospray ionization, Quadrupole and Time of Flight (TOF) analyzers, and argon as a collision gas.
Ginsenosides and flavonoids mentioned above were identified on both LC/UV and LC/MS spectra of these studied herbs compared with standards.
These marker components in Panax ginseng, Panax quinquefolinm, Citrus aurantium, Astragalus membranaceus and Hedysarum polybotrys have been assembled as fingerprints in LC/UV or LC/MS spectra of identifying Chinese herbs.

目　錄
中文摘要 i
英文摘要 iii
誌謝 v
目錄 vi
表目錄 ix
圖目錄 x
第一章 緒論 1
1.1 前言 1
1.2 研究背景 2
1.3 研究目的 3
第二章 文獻回顧 4
2.1 前言 4
2.2 分析方法回顧 5
2.3 藥材介紹 7
2.2.1 人參 7
2.3.2 西洋參 10
2.3.3 枳殼 13
2.3.4 蒙古黃耆 15
2.3.5 紅耆 18
2.4 藥材指標成分介紹 21
2.4.1 生曬參、紅參及西洋參 21
2.4.2 枳殼 22
2.4.3 蒙古黃耆、紅耆及市售黃耆 24
2.5 儀器原理介紹 25
2.5.1 超音波溶劑萃取法 25
2.5.2 高效液相層析法 27
2.5.3 液相層析串聯質譜儀 29
第三章 實驗材料與方法 34
3.1 實驗藥品 34
3.2 實驗器材 35
3.3 實驗大綱 37
3.4 實驗設備 38
3.4.1 萃取設備 38
3.4.2 分析設備 38
3.5 實驗步驟 40
3.5.1 藥材前處理 40
3.5.2 樣品萃取 40
3.5.3 測定方法 40
3.6 液相層析儀分析條件開發 42
3.7 質譜儀分析條件開發 43
3.8 檢量線配製 44
3.9蒙古黃耆、紅耆及市售黃耆藥材指紋雷達圖建立 45
第四章 結果與討論 46
4.1 生曬參、紅參及西洋參分析 47
4.1.1 生曬參、紅參及西洋參液相層析儀分析條件 47
4.1.2 生曬參、紅參及西洋參質譜儀分析條件 48
4.1.3 生曬參、紅參及西洋參指紋圖譜 49
4.2 枳殼分析 51
4.2.1 枳殼液相層析儀分析條件 52
4.2.2 枳殼質譜儀分析條件 53
4.2.3 枳殼指紋圖譜 54
4.2.4 枳殼指標成分圖譜 57
4.3 蒙古黃耆、紅耆及市售黃耆分析 60
4.3.1 蒙古黃耆、紅耆及市售黃耆液相層析儀分析條件60
4.3.2 蒙古黃耆、紅耆及市售黃耆質譜儀分析條件 61
4.3.3 蒙古黃耆緩衝溶液方法比較 62
4.3.4 蒙古黃耆、紅耆及市售黃耆指紋圖譜 65
4.3.5 蒙古黃耆、紅耆及市售黃耆指標成分圖譜 68
4.3.6 蒙古黃耆、紅耆及市售黃耆指標成分MSMS圖譜73
4.3.7 蒙古黃耆、紅耆及市售黃耆指標成分含量測定76
4.3.8 蒙古黃耆、紅耆及市售黃耆藥材指紋雷達圖建立78
第五章 結論 80
參考文獻 81
附錄
A 藥材指標成分圖譜 87
B 六批不同來源蒙古黃耆、紅耆及市售黃耆藥材分析光譜 101
C 六批不同來源蒙古黃耆、紅耆及市售黃耆藥材分析數據 108
D 指標成分結構 118

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