研究生: |
黃文盈 wen-ying Huang |
---|---|
論文名稱: |
中藥材隻組成成分分析與基原辨識 |
指導教授: |
許順吉
Xu, Shun-Ji |
學位類別: |
博士 Doctor |
系所名稱: |
化學系 Department of Chemistry |
論文出版年: | 2006 |
畢業學年度: | 94 |
語文別: | 中文 |
論文頁數: | 401 |
中文關鍵詞: | 連翹 、龍膽草 、分析方法開發 、基原鑑定 |
英文關鍵詞: | HPLC, LC-MS, ICP-MS, SPSS |
論文種類: | 學術論文 |
相關次數: | 點閱:404 下載:0 |
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化學分析是中藥科學化的重要內涵。本研究旨在開發連翹、龍膽草、川芎、當歸及防己藥材之HPLC分析方法,並藉由LC-MS方法確認或推測各成分的結構,然後將多批樣品的分析數據結果整理歸納,建立藥材基原與化學成分指紋圖譜間的關係。另外,不同植物含有不同種類與含量的微量元素,我們亦利用感應耦合電漿質譜儀(ICP-MS)分析連翹與龍膽草不同品種間的微量元素相對含量,並應用於藥材基原之化學辨識。
本研究分四個部分,第一部分為連翹成分之分析以及連翹藥材的化學基原鑑別。連翹具清熱、解毒、散結、消腫、排膿之功能,主要含rengyoside B (1), matairesinol (2), salidroside (3), quercitrin (4), suspensaside (5), rengyoside C (6), matairesinoside (7), forsythiaside (8), pinoresinol-β-D-glucoside (9), cornoside (10), acteoside (11), arctigenin (12), arctiin (13), phillyrin (14)及 pinoresinol (15)等成分。這些成分可用磷酸鹽、氰甲烷及甲醇水溶液為沖提液,254 nm為偵測波長的分離條件,在60分鐘內完成分析。該分析方法再現性為同一天內(n=6)面積比的相對標準偏差在0.23-1.5%之間,不同天面積比的相對標準偏差在0.41-2.40%之間,再現性良好。本研究進一步利用LC-MS以確認或推測連翹特殊波峰的化學結構,並以統計軟體搭配各種不同的數據處理方式,完成連翹藥材之數據化指紋圖譜分析。
第二部分為龍膽草成分之分析以及龍膽草藥材的化學基原鑑別。龍膽草具有除下焦溼熱、瀉肝膽實火之功能,主要含tetraacetylgentiopicroside (1), swertiamarin (2), 6β-hydroxyswertiajaposide (3), sweroside (4), swertiajaposide A (5), gentianine (6), gentiopicroside (7), gentiobiose (8), gentioside (9), gelidoside (10), olivieroside (11), loganic acid (12), gentianose (13), scabroside (14) 及amaroswerin (15)等成分。這些成分可用磷酸鹽、氰甲烷及甲醇水溶液為沖提液,254 nm為偵測波長的分離條件,在60分鐘內完成分析。該分析方法再現性為同一天內(n=6)面積比的相對標準偏差在0.33-1.65%之間,不同天面積比的相對標準偏差在0.68-2.67%之間,再現性良好。本研究進一步利用LC-MS以確認或推測龍膽草特殊波峰的化學結構,並以統計軟體搭配各種不同的數據處理方式,完成龍膽草藥材之數據化指紋圖譜分析。
第三部分為微量元素分析,分析連翹不同成熟度中青翹與老翹在微量元素組成上的差異,以及龍膽草藥材中龍膽草、條葉龍膽與堅龍膽的微量元素含量。實驗結果顯示,青翹與老翹的微量元素含量組成的差異,老翹中重要的微量元素要比青翹為高,而以Mg、Ca、Ru等元素作辨識指標,則可明確分辨樣品基原;取龍膽草作微量元素分析,發現多數元素含量皆以堅龍膽為最高,條葉龍膽為最低,而以Mg、K、Ge、Se、Rb、Nd等元素作辨識指標,則可明確分辨樣品基原。
第四部份為川芎、當歸與防己之成分分析。川芎與當歸均為常用中藥材,前者具有鎮靜、活血袪瘀、袪風止痛的功能,後者具鎮靜、活血、補血、調經等功能,都含有精油、胺基酸、脂肪酸、有機酸、糖類等成分;其中,有機酸以nicotinic acid (1)、phthalic acid (2)、protocatchuic acid (3)、folic acid (4)、p-hydroxybenzoic acid (5)、folinic acid (6)、vanillic acid (7)、caffeic acid (8)、ferulic acid (9)為主要組成。等成分。以HPLC進行分析,實驗結果顯示,利用磷酸鹽及氰甲烷水溶液為沖提液,以210nm為偵測波長,可在60分鐘內有效分析該9個有機酸成分,本研究並進一步利用LC-MS以確認此9個有機酸成分特殊波峰的化學結構。防己具有利水消腫、袪風止痛的功效,其內含acurumidine (1), magnoflorine (2), stepharine (3), sinomenine (4), acutumine (5), cyclanoline (6), fangchinoline (7), berbamine (8), tetrandrine (9), isotetrandrine (10), trilobine (11), aristolochic acid Π (12), aristolochic acid Ι (13), aristoloactam (14)及isotrilobine (15)等成分。以HPLC進行分析,實驗結果顯示,利用醋酸鹽、及氰甲烷水溶液為沖提液,以254nm為偵測波長,可在60分鐘內有效分析該15個生物鹼成分,本研究並進一步利用LC-MS以確認或推測其特殊波峰的化學結構。
Chemical analysis plays an important role for modernizing the Chinese herbal medicine. In this study, HPLC methods for analyzing Forsythiae Fructus, Gentianae scabrae Radix, Ligustuci Rhizoma, Angelicae Radix and Fangchi Radix were developed. LC-MS method for predicting or confirming the structures of each component were also developed. With analytical data from several groups of samples, the relationship between drug sources and fingerprints of chemical components were established. In addition, inductively coupled plasma mass spectrometer (ICP-MS) was used to analyze the amount of trace elements in different species of Forsythiae Fructus and Gentianae scabrae Radix. These results can be applied to identify the species of commercial samples.
There are four parts in this study. The first part is the component analysis and origin identification of Forsythiae Fructus. Forsythiae Fructus is usually applied to patients with dispels heat、remove toxin、disperses accumulations and swelling、drains pus and contains constituents rengyoside B (1), matairesinol (2), salidroside (3), quercitrin (4), suspensaside (5), rengyoside C (6), matairesinoside (7), forsythiaside (8), pinoresinol-β-D-glucoside (9), cornoside (10), acteoside (11), arctigenin (12), arctiin (13), phillyrin (14) and pinoresinol (15). Results showed that 15 components can be determined by HPLC in 60 minutes with a mobile phase containing phosphate, acetonitrile, and methanol at a detecting wavelength of 254 nm. The peak-area ratio reproducibility (RSD) of the proposed method was 0.23~1.50 % (intraday) and 0.41~2.40 % (interday). This study further confirmed or predicted the chemical structure of specific peaks obtained from Forsythiae Fructus by LC-MS and fingerprint analysis of Forsythiae Fructus was accomplished using the statistical software with some data processing methods.
The second part is the component analysis and origin identification of Gentianae scabrae Radix. Gentianae scabrae Radix is usually applied to patients with expels inflammation、dispels heat、purges excess fire in the liver and gallbladder、expels dampness-heat in the lower warmer and contains constituents tetraacetylgentiopicroside (1), swertiamarin (2), 6β-hydroxyswertiajaposide (3), sweroside (4), swertiajaposide A (5), gentianine (6), gentiopicroside (7), gentiobiose (8), gentioside (9), gelidoside (10), olivieroside (11), loganic acid (12), gentianose (13), scabroside (14) and amaroswerin (15). Results showed that 15 components can be determined by HPLC in 60 minutes with a mobile phase containing phosphate, acetonitrile, and methanol at a detecting wavelength of 254 nm. The peak-area ratio reproducibility (RSD) of the proposed method was 0.33~1.65% (intraday) and 0.68~2.67% (interday).This study further confirmed or predicted the chemical structure of specific peaks obtained from Gentianae scabrae Radix by LC-MS and fingerprint analysis of Gentianae scabrae Radix was accomplished using the statistical software with the data processing methods.
The third part is the analysis of trace elements. The content of trace elements in different Forsythiae Fructus and Gentianae scabrae Radix samples were analyzed. Results showed that the elemental profile of young and old Forsythiae Fructus are somewhat different. The old Forsythiae Fructus contains higher amount of important elements than young Forsythiae Fructus. And we could identify the Forsythiae Fructus samples by the recognized targets, Mg、Ca、Ru. For analyzing the trace elements in Gentianae scabrae Radix, the amount of most elements contained in G. regescens Franch is highest and in G. manshurica Kitag. is lowest. And we could identify the Gentianae scabrae Radix samples by the recognized targets, Mg、K、Ge、Se、Rb、Nd.
The fourth part is the components analysis and origin identification of Ligustuci Rhizoma、Angelicae Radix and Fangchi Radix. Ligustuci Rhizoma and Angelicae Radix are commonly used Chinese herb drugs. The former possesses tranquilizing, blood enlivening, hemostasis-resolving, anemofugic and analgesic effects; and the latter possesses tranquilizing, blood enlivening, hemopoietic and menstrual regulating effects. Both drugs contain essential oils, amino acids, fatty acids, organic acids and sugars. The organic acids compose chiefly of nicotinic acid (1)、phthalic acid (2)、protocatchuic acid (3)、folic acid (4)、p-hydroxybenzoic acid (5)、folinic acid (6)、vanillic acid (7)、caffeic acid (8) and ferulic acid (9). Results showed that 9 components can be determined by HPLC in 60 minutes with a mobile phase containing phosphate and acetonitrile at a detecting wavelength of 210 nm. This study further confirmed or predicted the chemical structure of specific peaks obtained from organic acids by LC-MS. Fangchi Radix item possess diuretic、detumescent、anemofugic and analgesic effects and contains constituents acurumidine (1), magnoflorine (2), stepharine (3), sinomenine (4), acutumine (5), cyclanoline (6), fangchinoline (7), berbamine (8), tetrandrine (9), isotetrandrine (10), trilobine (11), aristolochic acid Π (12), aristolochic acid Ι (13), aristoloactam (14) and isotrilobine (15). Results showed that 15 alkaloid components can be determined by HPLC in 60 minutes with a mobile phase containing phosphate and acetonitrile at a detecting wavelength of 254 nm. This study further confirmed or predicted the chemical structure of specific peaks obtained from Fangchi Radix by LC-MS.
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