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研究生:蔡輝隆
研究生(外文):Huei-Long Tsai
論文名稱:以高效液相層析儀定量分析黃芩及含黃芩處方煎劑之黃芩?研究
論文名稱(外文):Quantitative Analysis of Baicalin in Scutellaria baicalin Georgi and Herbal Prescriptions Containing Scutellariae Radix by HPLC
指導教授:蔡東湖蔡東湖引用關係王聲遠王聲遠引用關係
指導教授(外文):Tung-Hu Tsai, Ph.D.Sheng-Yuan Wang M.D.
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:傳統醫藥學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:80
中文關鍵詞:高效液相層析儀黃芩黃芩?
外文關鍵詞:HPLCScutellaria baicalin GeorgiBaicalin
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中 文 摘 要
 
黃芩?是唇形科多年生草本植物黃芩的根所萃取的有效黃酮類,有抗生素、抗氧化、治療肝癌等多重生物效能。傳統中藥黃芩,幾千年已普遍用於治療。為了要建立一套高效液相層析法來偵測含黃芩?藥材,探討含黃芩複方煎劑之特性,本實驗高效液相層析系統,包含層析用幫浦,配備10 μl檢品注射器,紫外線偵測器。逆相層析碳18管柱,內徑150x1 mm,質粒大小5 μm,移動相是10 mM磷酸二氫鈉與乙氰胺混合液(31:69,v/v)用磷酸調至pH 3.0,流速0.05 ml/min,紫外線檢測波長設在最大吸光值的277 nm。為確認檢品波峰,進行五項實驗:與標準溶液比較;與不含黃芩之空白溶液比較;額外加入黃芩?標準溶液;紫外線檢測波長由277變成300 nm;移動相組成條件,加5 %THF的改變。同日及不同日準確性與精密度皆在可接受範圍內。偵測極限為50 ng/ml,黃芩?的檢量線在濃度0.05-50 μg/ml內呈線性關係。平均實驗流程回收率是96.23 ±0.46 %。實驗由簡而繁,由煮沸次數、煮沸時間、溶媒的決定起,試驗不同炮製方式,測試與其它藥材配伍相生相剋。進而擴大到十個複方。其中黃連解毒湯、小柴胡湯和龍膽瀉肝湯,黃芩與之一對一、一對二、一對三等各個之間測試交互作用。再擴展到臨床特殊療效分析,整個實驗結構力求完整。戲劇性的發現:黃芩單獨使用,黃芩?萃取量最高,水溶液是84.15 ±1.04 mg/g;50 %乙醇溶液是145.14 ±8.74 mg/g;黃芩與其它藥材一起使用,黃芩?萃取量有統計意義的減少。其機制尚未釐清,有待進一步探討。結果顯示:高效液相層析法偵測黃芩及含黃芩藥材,沒有干擾且足夠靈敏以偵測之。黃芩及含黃芩處方煎劑蘊涵藥藥之交互作用。

Abstract
Scutellaria baicalensis Georgi, a traditional Chinese medicine, has been used as a popular remedy for thousands of years. Baicalin, one of the bioactive flavonoids isolated from Scutellariae Radix, has been used as antibiotics, antioxidant, against hepatic fibrosis, carcinoma etc. In order to develop a HPLC method for the determination of baicalin from herb medicine, to characterize the baicalin content in the traditional prescriptions. This study designed the liquid chromatographic system consisted of a chromatographic pump, an injector equipped with a 10 μl sample loop and a Dynamax UV/VIS detector. Samples were separated on a reversed-phase C18 column (150 x 1 mm I.D.; particle size 5 μm), eluted with a mobile phase consisted of 10 mM monosodium phosphoric acid - acetonitrile (69:31, v/v, pH 3.0) with a flow rate 0.05 ml/min. The UV detector wavelength was set at 277 nm. Five quality experiments were assayed to identify the peak of sampling which include compared with standard baicalin, compared with blank excluding Scutellariae Radix, extra-addition of authentic standard baicalin, changing wavelength from 277 to 300 nm, changing the composition of mobile phase with 5 % THF. Then, intra-assay, inter-assay accuracy and precision of baicalin levels fell well within the predefined limits of acceptability in the range of 0.05 through 50 μg/ml. The limit of quantification was 50 ng/ml. The extraction processing recovery of baicalin was 96.2 ± 0.46 %. The samples were processed by the following steps:One time extraction of baicalin, two different solvents, five boiling intervals, five prepared processes, five compatibility applications, ten the most popular herbal prescriptions containing Scutellariae Radix in Taiwan, including mutual interaction in detail individually. Finally, we extended the quantitative analysis to the special clinical use of Scutellariae Radix. Dramatically, the highest baicalin extraction was 84.15 ± 1.04, 145.14 ± 8.74 mg/g in water and in 50 % ethanol respectively while Scutellariae Radix was used lonely. However, the baicalin extraction was significantly decreased while Scutellariae Radix was used combined with others. It is worthy to research more the unknown mechanism. The results indicated that no other ingredients would interfere significantly for the determination of the baicalin in this chromatographic analytic system. This HPLC method is sufficiently sensitive to allow measurement of baicalin in medicinal herbs and their pharmaceutical products. Ingredient-ingredient interactions were involved in traditional Chinese medicines and the related herbal prescriptions

目 錄
目錄……………………………………………………………………… 1
表目錄…………………………………………………………………… 3
圖目錄…………………………………………………………………… 4
中文摘要………………………………………………………………… 5
英文摘要………………………………………………………………… 6
第一章 緒論…………………………………………………………… 8
壹、研究動機………………………………………………… 10
貳、簡介黃芩及黃芩?……………………………………… 12
參、黃芩及黃芩?之藥理作用…………………………… 14
第二章 研究目的……………………………………………………… 18
第三章 材料與方法…………………………………………………… 20
壹、儀器設備………………………………………………… 22
貳、試藥來源與配備………………………………………… 23
參、實驗設計………………………………………………… 24
肆、統計……………………………………………………… 29
第四章 結果…………………………………………………………… 30
壹、準確性與精密度………………………………………… 32
貳、實驗流程回收率………………………………………… 32
參、實驗檢品層析圖波峰之確認…………………………… 32
肆、不同溶媒對黃芩?萃取量之影響……………………… 33
伍、煮沸時間對黃芩?萃取量之影響……………………… 34
陸、第一次抽取與第二次抽取對黃芩?
萃取量影響之比較……………………………………… 34
柒、炮製方法對黃芩?萃取量之影響……………………… 34
捌、配伍國老對黃芩?萃取量之影響……………………… 35
玖、配伍相使對黃芩?萃取量之影響……………………… 35
拾、配伍相畏對黃芩?萃取量之影響……………………… 36
拾壹、黃芩?在含有黃芩處方中之萃取量………………… 36
拾貳、黃連解毒湯各成份藥材對黃芩?萃取量之影響… 36
拾參、小柴胡湯各成份藥材對黃芩?萃取量之影響……… 37
拾肆、龍膽瀉肝湯各成份藥材對黃芩?萃取量之影響…… 37
拾伍、配伍柴胡對黃芩?萃取量之影響…………………… 37
拾陸、臨床用藥對黃芩?萃取量之影響…………………… 38
第五章 討論…………………………………………………………… 39
第六章 結論…………………………………………………………… 50
第七章 參考文獻……………………………………………………… 52
附錄表………………………………………………………………… 57
附錄圖………………………………………………………………… 64
表 目 錄
表1、Assay validation of baicalin…………………………………… 58
表2、Recovery of baicalin extraction processing…………………… 58
表3、Effect of solvent to the baicalin extraction……………………… 59
表4、Comparison of twice extracting baicalin……………………… 59
表5、Effect of prepared processing to the baicalin extraction……… 60
表6、Effect of Glycyrrhizae Radix to the baicalin extraction……… 60
表7、Effect of compatibility applications to the baicalin extraction…… 61
表8、 Baicalin extraction of the prescriptions contained Sutellariae
Radix………………………………………………………… 61
表9、Interaction of Huang-Lian-Jiee-Dwu-Tong contents to baicalin extraction 62
表10、Interaction of Long-Dan-Xie-Gan-Tong contents to baicalin extraction 63
表11、Interaction of clinical use drug to the baicalin extraction………… 63
圖 目 錄
圖1、Chemical structure of baicalin………………………………… 65
圖2、UV Spectrum of baicalin……………………………………… 66
圖3、Chromatogram of changing UV wavelength…………………… 67
圖4、Chromatogram of changing mobile phase composition……… 68
圖5、Chromatogram of excluding Scutellariae Radix as a blank…… 69
圖6、Chromatogram of adding standard biacalin…………………… 70
圖7、Effect of boiling interval to the baicalin extraction……………… 71
圖8、Chromatogram of prepared processes…………………………… 72
圖9、Effect of compatibility applications incompatible herb
to baicalin extraction…………………………………………… 73
圖10、Efect of Xiao-Chai-Hu-Tong contents to baicalin extraction… 74
圖11、Chromatogram of herbal prescriptions containing
Scutellariae Radix……………………………………………… 75

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