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研究生: 盛珮雯
Pei-Wen Sheng
論文名稱: 中藥製劑之毛細管電泳及高效液相層析研究
指導教授: 許順吉
Xu, Shun-Ji
學位類別: 碩士
Master
系所名稱: 化學系
Department of Chemistry
論文出版年: 2000
畢業學年度: 88
語文別: 中文
論文頁數: 143
中文關鍵詞: 毛細管電泳高效液相層析加味逍遙散掃集法中藥製劑
英文關鍵詞: CE, capillary electrophoresis, HPLC, high performance liquid chromatography, sweeping, Chinese herbal preparation, MEKC, rev. EOF CZE
論文種類: 學術論文
相關次數: 點閱:393下載:0
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  • 高效液相層析(HPLC)和毛細管電泳(CE)同屬液相分離技術,都有多種分離模式,為目前最常用來定量中藥材指標成分的分析方法。本研究以這兩種儀器開發加味逍遙散及其他中藥製劑的分析方法,並比較HPLC和CE兩分析方法的適用性。
    加味逍遙散是由當歸、白芍、白朮、茯苓、柴胡、牡丹皮、山梔子、炙甘草、生薑、薄荷等十種藥材所組成,在HPLC分析中,沖提系統之流動相(A)為30mM KH2PO4 (以10%H3PO4調整pH至2.90),流動相(B)為CH3CN/H2O = 85/15(V/V),利用Cosmosile 5C18-MS之分離管柱,以203、210及254 nm為偵測波長,利用梯度沖提系統可在70分鐘內分析加味逍遙散的二十個指標成分(trans-ferulic acid、cis-ferulic acid、nicotinic acid、phthalic acid、cinnamic acid、folinic acid.、benzoic acid、gallic acid、paeonol、paeoniflorin、albiflorin、oxypaeoniflorin、saikosaponin a、geniposide、geniposidic acid、shanzhiside methylester、glycyrrhizin、18b-Glycyrrhetinic acid、6-gingerol及menthol)。本文另以此分析法評估加味逍遙散科學製劑的安定性。
    在CE分析中,根據加味逍遙散中各指標成分之不同性質而分成兩部分進行,即MEKC及rev. EOF CZE。在MEKC部分,利用含陰離子界面活性劑(SC和SDS)和有機修飾劑之緩衝溶液,可在35分鐘內分析其中九種指標成分;在rev. EOF CZE部分,利用含陽離子界面活性劑(CTAB)和有機修飾劑之緩衝溶液,可在10分鐘內分析其中九種有機酸指標成分。該分析條件之選擇和評估亦一併探討。此外,以一種線上堆積法(Sweeping)進行柴胡及其相關製劑的分析,並比較此方法和傳統CE分析方法的差異和優劣。
    檢討加味逍遙散之HPLC和CE的分析結果,發現在系統適宜性(包含再現性、回收率和偵測極限)、分析時間、基線穩定性及各吸收峰之理論板數等方面各有其優劣點,兩者應可相輔相成、搭配使用。

    High-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) are two well-known technologies in the field of chemical analysis. Both have many separating modes and are used to assay the Chinese herbal drugs. In this study, two analytical methods basing on these technologies were established to assay the constituents of one Chinese herbal preparation, Chia-wei-hsiao-yao-san, and related Chinese herbal drugs. The suitability of the two methods was also evaluated.
    Chia-wei-hsiao-yao-san comprises Angelicae Radix, Paeoniae Radix, Atractylodis Rhizoma, Poria, Bupleuri Radix, Moutan Radicis Cortex, Gardeniae Fructus, Glycyrrhizae Radix, Zingiberis Rhizoma, and Menthae Herba. In HPLC method, twenty constituents including trans-ferulic acid, cis-ferulic acid, nicotinic acid, phthalic acid, cinnamic acid, folinic acid., benzoic acid, gallic acid, paeonol, paeoniflorin, albiflorin, oxypaeoniflorin, saikosaponin a, geniposide, geniposidic acid, shanzhiside methylester, glycyrrhizin, 18b-Glycyrrhetinic acid, 6-gingerol, and menthol were determined in 70 minutes. The analysis was achieved by using a Cosmosil 5C18-MS column with a linear gradient elution system containing phosphates and acetonitrile and detecting at 203 nm, 210 nm, and 254 nm. Additionally, the preservation stability of the commercial available Chia-wei-hsiao-yao-san was also evaluated by the developed method.
    In CE method, two modes, MEKC and rev. EOF CZE, were used according to the properties of the constituents. In MEKC, nine constituents could be separated in 35 minutes using a buffer system which contained anionic surfactants (SC and SDS) and a organic modifier (acetonitrile). In rev. EOF CZE, nine constituents belonging to organic acids could be separated in 10 minutes using a buffer system which contained a cationic surfactant (CTAB) and a organic modifier (acetonitrile). The effects of the analysis parameters were also investigated herein. Moreover, the Bupleuri Radix and relating preparations were analyzed by another technique, on-line sample stacking CE method (sweeping). The suitability and effects of sweeping method with those of the conventional CE methods were also compared.
    From the results, it was found that HPLC and CE possessed its own beneficial effects in terms of the suitability of the system, analyzing time, stability of the base line, and the number of theoretical plates of each compound, respectively. HPLC and CE can complement each other in the analysis of Chinese herbal preparations.

    圖目錄…………………………………………………………………….II 表目錄…………………………………………………………………….IV 中文摘要…………………………………………………………………VII 英文摘要……………………………………………………………….…IX 第一章 緒論 I - 1 前言…………………………………………………………..1 I - 2 高效液相層析法……………………………………………..3 I - 3 毛細管電泳分析法…………………………………………..5 I - 4 分析條件參數與適宜性之評估……………………………18 第二章 加味逍遙散及相關中藥製劑之高效液相層析 II - 1 前言…………………………………………………………23 II - 2 實驗部分……………………………………………………33 II - 3 結果與討論…………………………………………………39 II - 4 加味逍遙散濃縮製劑之安定性評估………………………56 第三章 加味逍遙散及相關中藥製劑之高效液相層析 III - 1 前言…………………………………………………….……65 III - 2 加味逍遙散之rev. EOF CZE分析…………………………65 III - 3 加味逍遙散之MEKC分析…………………………………90 III - 4 樣品堆積法Sweeping之分析試驗及討論………………..106 第四章 結論 IV - 1 HPLC與CE之比較……………………………………….122 IV - 2 結論………………………………………………………...128 參考文獻…………………………………………………………………131 附 錄 加味逍遙散指標成分之UV吸收光譜圖…………………….137

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