研究生: |
李淑玲 shu-ling li |
---|---|
論文名稱: |
益氣聰明湯之CE與HPLC分析 The analysis of Yi-chi-tsung-ming-tang in CE and HPLC |
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: | 128 |
中文關鍵詞: | 益氣聰明湯 、CE 、HPLC |
英文關鍵詞: | Yi-chi-tsung-ming-tang, CE, HPLC |
論文種類: | 學術論文 |
相關次數: | 點閱:258 下載:79 |
分享至: |
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報 |
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益氣聰明湯的分析方法,並比較HPLC和CE兩分析方法的適用性。
益氣聰明湯是由白芍、黃耆、人參、葛根、蔓荊子、升麻、甘草、黃柏等八種藥材所組成,在HPLC分析中,沖提系統之流動相A為20mM KH2PO4 (以10%H3PO4調整pH至3.74),流動相B為CH3CN/H2O = 80/20(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80分鐘內分析益氣聰明湯的十七個指標成分(gallic acid、benzoic acid、oxypaeoniflorin、albiflorin、paeoniflorin、paeonol、formononetin、ferulic acid、caffeic acid、puerarin、daidzein、berberine、palmatine、jatrorrhizin、glycyrrhizin、18b-glycyrrhetinic acid及ginsenoside Rb1)。本文另以此分析法探討益氣聰明湯中各藥材與所成方劑之間各成分含量的消長關係,並藉此分析法評估益氣聰明湯濃縮製劑保存的安定性。
在CE分析中,根據益氣聰明湯中各指標成分之不同性質而分成兩部分進行,即MEKC及CZE。在MEKC部分,利用含陰離子界面活性劑(50 mM SC)和有機修飾劑(氰甲烷)之緩衝溶液,可在15分鐘內分析其中albiflorin、paeoniflorin兩種指標成分;在CZE部分,硼酸鹽(pH=9.76)之緩衝溶液,可在55分鐘內分析其中十三種指標成分。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)進行茵陳蒿的分析,並比較此方法和傳統CE分析方法的差異和優劣。
檢討益氣聰明湯之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勢,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 techniques in chemical analysis. Both have many separating modes and are used to assay Chinese herbal drugs. In the present study, two analyzing methods are established for assaying the constituents of the Chinese herbal preparation, Yi-chi-tsung-ming-tang. The suitability of the two methods was also evaluated and compared.
Yi-chi-tsung-ming-tang comprises Paeoniae Radix, Astragali Radix, Ginseng Radix, Puerariae Radix, Viticis Fructus, Cimicifugae Rhizoma, Glycyrrhizae Radix and Phellodendri Cortex. In HPLC analysis, eighteen constituents of Yi-chi-tsung-ming-tang, including gallic acid, benzoic acid, oxypaeoniflorin, albiflorin, paeoniflorin, paeonol, formononrtin, ferulic acid, caffeic acid, puerarin, daidzein, berberine, palmatine, jatrorrhizin, glycyrrhizin, 18b-glycyrrhetinic acid and ginsenoside Rb1, were separated in 80 minutes by a Cosmosil 5C18-MS column with a linear gradient elution system containing phosphates and acetonitrile which was detected at 203 nm, 210 nm, and 254 nm. Moreover, the constituents in each component herb and the home-made preparation were investigated herein. The preservation stability of the commercially available Yi-chi-tsung-ming-tang preparations was also evaluated by the developed analysis method.
In CE analysis, the analysis of Yi-chi-tsung-ming-tang was cut into two parts, MEKC and CZE, according to the properties of the constituents In MEKC, two constituents, albiflorin and paeoniflorin, could be separated in 15 minutes, using a buffer system containing anionic surfactants (40 mM SC) and acetonitrile as organic modifier. In CZE, thirteen constituents could be separated in 55 minutes, using a buffer system containing 20 mM borate(pH=9.76). The effects of the analysis parameters were also investigated herein. Moreover, one Chinese herb, Artemisiae Capillaris Herba, was analyzed by an on-line sample stacking CE method, sweeping. Also, the suitability and effects of the method were compared with those of the conventional CE methods.
From the results of the analysis, it is concluded that HPLC and CE possesses their 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.
[1] 何志鋒,《中國醫學綱要》,第3頁,國立中國醫藥研究所,台北,1978。
[2] 許鴻源,《中國醫學概念》,第1-19頁,行政院衛生署中醫藥委員會,台北,1973。
[3] M. Tweet, Proc. Warsaw Soc. Nat. Sci. Biol., 1903, 14, 6.
[4] A. J. P. Martin and R. L. M Synge, Biochem. J., 1941, 35, 1358.
[5] A. T. James and A. J. P. Martin, Analyst, 1952, 77, 915.
[6] E. Stahl, Chemiker-Ztg., 1958, 82, 323.
[7] J. F. K. Huber and J. A. R. J. Hulsman, Anal. Chem. Acta, 1967, 38, 305.
[8] A. Tiselius, Tran. Faraday Soc., 1937, 33, 524.
[9] S. Hjertén, Chromatogr. Rev., 1967, 9, 122.
[10] R. Virtanen, Acta Polytechnica Scand., 1974, 123, 1.
[11] F. E. P. Mikkers, F. M. Everaerts, and T. P. E. M. Verheggen, J. Chromatogr., 1979, 169, 11.
[12] J. W. Jorgenson and K. D. Lukacs, Anal. Chem., 1981, 53, 1298.
[13] S. Hjertén, J. Chromatogr., 1985, 347, 171.
[14] J. R. Mazzeo and I. S. Krell, BioTechniques, 1991, 10, 638.
[15] T. Wehrpursue, LC-GC Mag., 1993, 11, 14.
[16] A. S. Cohen and B. L. Karger, J. Chromatogr., 1987, 397, 409.
[17] A. S. Cohen, A. Paulus, and B. L. Karger, Chromatographia, 1987, 24, 15.
[18] A. S. Cohen, D. R. Najarian, A. Paulus, A. Guttman, J. A. Smith, and B. L. Karger, Proc. Natl. Acad. Sci. U.S.A., 1988, 85, 9660.
[19] H. Drossman, J. A. Luckey, A. Kostichka, J. D’Cunha, and L. M. Smith, Anal. Chem., 1990, 62, 900.
[20] P. Bocek and A. Chrambach, Electrophoresis, 1991, 12, 1059.
[21] S. Terabe, K. Otsuka, K. Ichikawa, A. Tsuchiya, and T. Ando, Anal. Chem., 1984, 56, 111.
[22] S. Terabe, K. Otsuka, and T. Ando, Anal. Chem., 1985, 57, 834.
[23] S. Hjertén, J. L. Liao, and K. Yao, J. Chromatogr., 1987, 387, 127.
[24] D. J. Rose and J. W. Jorgenson, Anal. Chem., 1988, 60, 642.
[25] T. Tsuda, Handbook of Capillary Electrophoresis; Landers, J. P., Ed.; CRC Press 1997; Chapter 1.
[26] H. Z. Helmholtz, Annal. Phys. Chem., 1879, 7, 337.
[27] 許鴻源、許照信,《常用漢方方劑圖解》,第199-200頁,新醫藥出版社,台北,1985。
[28] 許鴻源、陳玉盤、許順吉、許照信、陳建志、張憲昌,《簡明藥材學》,第402-404頁,新醫藥出版社,台北,1985。
[29] 許鴻源,《中藥材之研究》,第121-123頁,新醫藥出版社,台北,1980。
[30] 同[28],第407-408頁。
[31] 同[29],第71-75頁。
[32] 同[28],第52-54頁。
[33] 同[29],第124-126, 567, 725, 732-733頁。
[34] 同[28],第55-56頁。
[35] 同[29],第87-88頁。
[36] 同[28],第421頁。
[37] 同[29],第7-10頁。
[38] 同[28],第117-118頁。
[39] 同[29],第88-93頁。
[40] 同[28],第47-48頁。
[41] 同[29],第161-163頁。
[42] 同[28],第410-411頁。
[43] 同[29],第126-130, 506-507, 530-532頁。
[44] 林文中,國立台灣師範大學化學研究所碩士論文,台北,1995。
[45] 吳心楷,國立台灣師範大學化學研究所碩士論文,台北,1997。
[46] D. N Heiger, “High Performance Capillary Electrophoresis – An Introduction”,p. 62, Hewleet-Packard GmbH, Waldbronn, 1992.
[47] D. S. Craig, J. C. Y. Wong, N. J. Dovichi,Anal. Chem., 1996, 68, 697.
[48] F. E. P. Mikkers, F. M. Everaerts, and T. P. E. M. Vertheggen, J. Chromatogr., 1979, 169, 11.
[49] S. E. Moring, J. C. Colburn, P. D. Grossman, H. H. Lauer, LC-GC Intl., 1990, 3, 46.
[50] R. L. Chien and D. S. Burgi, Anal. Chem., 1992, 64, 489A.
[51] D. S. Burgi and R. L. Chien, Anal. Chem., 1992, 63, 2042A.
[52] D. S. Burgi and R. L. Chien, J. Microcol. Sep., 1991, 3, 199.
[53] D. S. Burgi and R. L. Chien, Anal. Biochem., 1992, 202, 306.
[54] R. L. Chien and D. S. Burgi, J. Chromatogr., 1991, 559, 141.
[55] R. L. Chien and D. S. Burgi, J. Chromatogr., 1991, 559, 143.
[56] Z. Liu, P. Sam, S. R. Sirimanne, P. C. McClure, J. Chromatogr. A, 1994, 673, 125.
[57] J. P. Quirino and S. Terabe, J. Chromatogr. A, 1997, 781, 119.
[58] J. P. Quirino and S. Terabe, J. Chromatogr. A, 1997, 791, 225.
[59] J. P. Quirino and S. Terabe, Anal Chem., 1998, 70, 149.
[60] K. Otsuka and S. Terabe, J. Microcol. Sep., 1998, 1, 150.
[61] J. P. Quirino and S. Terabe, J. Chromatogr. A, 1998, 798, 251.
[62] J. P. Quirino and S. Terabe, Anal Chem., 1998, 70, 1893.
[63] J. P. Quirino and S. Terabe, J. Chromatogr. B, 1998, 714, 29.
[64] J. P. Quirino and S. Terabe, Science, 1998, 282, 465.
[65] 簡君玲,國立台灣師範大學化學研究所碩士論文,台北,1998。
[66] 同[28],第209-210頁。
[67] 同[29],第75-78, 538頁。
[68] 江蘇新醫學院編,《中藥大辭典》下冊,第1588頁,上海科學技術出版社,上海1978。
[69] 宋 唐慎微,《重修改正和經史証類備用本草》,第七卷,第188頁,人民衛生出版社,北京1978。
[70] 李桂蘭,國立台灣師範大學化學研究所碩士論文,台北,1995。
[71] N. Asakawa, T. Hattori, M. Ueyama, A. Shinoda and Y. Miyake, Yakuguku Zasshi, 1979, 99, 598.
[72] Y. Katoh and T. Hayashi, Hokkaidorritsu Eisei Kenkyushoho., 1985, 35, 95; CA., 1985, 104, 182648p.
[73] 莊武璋,國立台灣師範大學化學研究所博士論文,台北,1996。
[74] 詹逸洲,國立台灣師範大學化學研究所碩士論文,台北,1993。
[75] 陳鴻仁,國立台灣師範大學化學研究所碩士論文,台北,1996。