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  • Theses

益氣聰明湯之CE與HPLC分析

The analysis of Yi-chi-tsung-ming-tang in CE and HPLC

Advisor : 許順吉教授
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Abstracts


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。

Keywords

益氣聰明湯 CE HPLC

Parallel abstracts


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.

Parallel keywords

Yi-chi-tsung-ming-tang CE HPLC

References


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