高效液相層析（HPLC）及毛細管電泳（CE）是目前最常用來測定中藥成分含量的分析方法。綜合兩者的優點，合併運用，可以拓展中藥化學評價的範疇。
本研究分三個部分，第一部分為生體介質中黃連成分之測量，我們採用分析黃連的CE方法，在22分鐘內完成三黃瀉心湯中的epiberberine、columbamine、berberastine、jatrorrhizine、berberine、palmatine、coptisine和magnoflorine等8個黃連生物鹼含量的測定，以探討三黃瀉心湯製劑在人工胃液、腸液中各成分含量的變化。結果發現黃連成分統計趨勢圖在人工胃液中幾不變動，但人工腸液的水層萃出物則和三黃瀉心湯組成藥材配伍之改變有關，當配伍中的大黃/黃芩比例增加時，黃連成分含量有下降趨勢，該現象可能來自溶解度及錯合沈澱物生成。另外，檢測餵食不同萃取方式的三黃浸膏之老鼠血清及尿液，發現在餵食24小時後的尿液才能用CE偵測得到，血清則偵測不到。
第二部分為開發HPLC分析方法，以偵測小白鼠尿液、血清中之稀薄黃連成分。本研究採梯度沖提的HPLC系統，流動相（A）為10mM KH2PO4 酸性水溶液（以10％H3PO4調整pH至2.63），（B）為CH3CN/H2O = 70/30（V/V）；小白鼠尿液、血清用氰甲烷萃取、濃縮，於80分鐘內可完成epiberberine、coptisine、berberine和palmatine等四個黃連生物鹼成分的分析。至於columbamine、jatrorrhizine 、berberastine和magnoflorine之吸收峰則與黃芩成分重疊，無法定量。本文比較三黃藥材個別煎煮、50％乙醇萃取和三黃藥材混和煎煮之三種浸膏樣品在動物體內的吸收代謝差異，結果發現不論餵食那一種浸膏，其尿液皆在12小時後，才呈現明顯黃連成分吸收峰；其中餵食混煎三黃浸膏之老鼠的尿中濃度含量最高。另外從各成份偵測值趨勢變化圖可發現餵食不同三黃浸膏之老鼠，各成份在血清中存留時間都很長。
第三部分為開發薑黃藥材之HPLC分析方法，薑黃為薑科植物薑黃 Curcuma longa Linn.的乾燥根莖，具有抗炎、抗菌、抗氧化、抗腫瘤、降血脂等作用。本研究採梯度沖提系統，流動相（A）10mM KH2PO4 酸性水溶液（以1％H3PO4調整pH至3.30），（B）CH3CN/H2O = 80/20（V/V），在分離管柱Cosmosile 5C18-MS、偵測波長254及280 nm的條件下，於40分鐘內完成分析薑黃藥材的α-turmerone、β-turmerone、ar-turmerone、 curcumin、demethoxycurcumin、bisdemethoxycurcumin、curcumenol和germacrone-13-al等八個指標成分。實驗結果顯示，本LC方法再現性良好，同一天與不同天滯留時間相對標準偏差分別在0.53-1.12%與0.02-0.69%之間，各成分回收率在96.8-104.2%之間。本研究另開發LC-MS分析方法，使用流動相(A)0.025% trifluoroacetic acid (TFA)，(B)MeOH/CH3CN/0.025%TFA = 50/30/20（V/V），可以分離α-turmerone與β-turmerone，用LC-MS的ESI+離子的模式可以確認各波峰成分。

Abstract
High-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) are currently the most commonly used analysis methods for determining the contents of constituents in Chinese herb drugs. By combining the superiorities of both methods, we can expand the scope of chemical appraisal of Chinese herb drugs.
This study is divided into three parts. The first part deals with the determination of Coptidis Rhizoma’s constituents in a living body. We adopted the CE method that has been used for the analysis of Coptidis Rhizoma. Within 22 minutes, we finished the determination of the eight alkaloids of Coptidis Rhizoma, namely epiberberine, columbamine, berberastine, jatrorrhizine, berberine, palmatine, coptisine and magnoflorine. Thereby we investigate the variation with the various constituents of the herb formula Coptis and Rhubarb Combination in artificial gastric fluid and artificial intestinal fluid. As a result, we found the contents of constituents did not change in artificial gastric fluid, but in the aqueous layer extracted from the artificial intestinal fluid, they changed someway related to the change in the composition of the formula. For example, as the rhubarb/scute ratio of the formula increases, the contents of the constituents of Coptidis Rhizoma displayed a decline tendency. Such a phenomenon could be attributable to solubility and formation of complexed precipitates . Besides, as mice were fed with the extracts of the formula extracted by different methods and the mice’s urine and serum were detected, it was found that only 24 hours after feeding can alkaloid constituents in the urine samples be detectable by CE, while serum samples were undetectable.
The second part is about the development of HPLC methods for detecting the trace amount contents of Coptidis Rhizoma constituents in the mouse’s urine and serum. The study applied the gradient elution system of HPLC with the mobile phases (A) composed of 10 mM KH2PO4 in an acid solution (adjusted to pH 2.63 with 10% H3PO4) and (B) composed of CH3CN/H2O = 70/30 (v/v). The mouse’s urine and serum was extracted with acetonitrile, concentrated and analyzed within 80 min for the four alkaloids of Coptidis Rhizoma: epiberberine, coptisine, berberine and palmatine. As for the constituents columbamine, jatrorrhizine, berberastine and magnoflorine, their absorption peaks overlapped with those of the constituents of Scutellariae Radix, and were undetectable. This study made a comparison of different extracts of the formula extracted with the ingredients of the formula decocted individually, with 50% ethanol and with the ingredients decocted together, which were fed to mice and their absorption and metabolism within the mice were compared. As a result, it was found that regardless of whichever extract, the peaks of the Coptidis Rhizoma constituents in the urine appeared conspicuously 12 hours later. Among the three extract, that extracted with the ingredients decocted together had the highest contents found in the urine. Also from the chart that showed the tendency of change of the various constituents detected, we found that the constituents of the three extracts all lingered in the serum for a very long time.
The third part of study covers HPLC analyses for Curcuma Tuber. Curcuma Tuber is the dried tuber of the Zingiberaceous plant Curcuma longa Linn, which possesses antiphlogistic, antimicrobial, antioxidation, anticancer and hypoglycemic effects. This study proposed a gradient elution system, with mobile phase (A) comprising 10 mM KH2PO4 in an acid solution (adjusted to pH 3.30 with 1% H3PO4) and (B) CH3CN/H2O = 80/20 (v/v). Using a separating column of Cosmosile 5C18-MS, under detecting wavelengths 254 and 280 nm, and within 40 min, we accomplished analysis of the eight marker substances for Curcuma Tuber, which were α-turmerone, β-turmerone, γ-turmerone, curcumin, demethoxycurcumin, bisdemethoxycurcumin, curcumenol and germacrone-13-al. The experimental results show that LC has very good reproducibility. Retentions times on the intraday day and on interday days differ by only 0.53 –1.12% and 0.02 – 0.69% respectively. The recovery rates of the various constituents are within 96.8 – 104.2%. This study also has developed an LC-MS analysis method using the mobile phase (A) 0.025% trifluoroacetic acid (TFA) and (B) MeOH/CH3CN/0.025%TFA = 50/30/20 (v/v), which can separate α-turmerone and β-turmerone. The peaks have been identified with the ESI+ ionization mode of LC-MS.

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