第一部份：以微乳液電動層析法分析大黃中之蒽醌類 (anthraquinones)及雙蒽酮類 (bianthrones) 之研究 第二部分：以毛細管區帶電泳法進行番瀉製劑中主要成分番瀉苷A (sennoside A) 與番瀉苷B (sennoside B) 之定性與定量分析之研究

Abstract

第一部份：以微乳液電動層析法分析大黃中之蒽醌類 (anthraquinones) 及雙蒽酮類 (bianthrones) 之研究 微乳液電動層析法與膠束電動層析法的分離原理相似，但是與膠束電動層析法相比較，它有較多的參數可調整用來改善分離結果。本研究是利用微乳液電動層析法對於九種蒽醌類及雙蒽酮類作分析方法開發的探討，藉由討論界面活性劑的比例、油相的種類及有機溶媒的添加，找出以0.5% (w/w) 的di-n-butyl-L-tartrate，1.2% (w/w) 的1-butanol, 0.6% (w/w) 的SDS，97.7% (w/w) 10 mM之硼酸鹽 (pH值9.2) 組成之微乳液，此外另添加30% (v/v) ACN，此條件能夠在20分鐘內將九種蒽醌類及雙蒽酮類完全分離。 因為考慮大黃中的成分複雜，所以我們採用固相萃取法當作樣品的前處理步驟，透過以弱酸、弱鹼及適當比例的有機溶媒對Oasis HLB管柱做樣品的充填與管柱的沖洗及沖提，將大黃粗萃取物做初步的分離，再使蒽醌類及雙蒽酮類以最適合之濃度注入毛細管電泳儀中進行定量。 第二部分：以毛細管區帶電泳法進行番瀉製劑中主要成分番瀉苷A (sennoside A) 與番瀉苷B (sennoside B) 之定性與定量分析之研究 本研究是利用毛細管區帶電泳對於市售的番瀉錠劑中具有瀉下作用，且含量最多的sennoside A及sennoside B做定性與定量的分析。 在分析方法開發的過程中，對於可能影響分離結果的參數做了一連串的探討，包括緩衝溶液的種類、濃度及pH值，分離時的溫及所施加的電壓與有機溶媒的添加等。經過多次嘗試，最後找出以100 mM 且pH值為10.4的CAPS緩衝溶液，溫度為30ºC，施加電壓為25 kV為最適化的分離條件，但是為了避免此次實驗所選定的內標準物與其他微量成分互相干擾，將此條件調整成以120 mM 且pH值為10.25的CAPS緩衝液，溫度為25ºC，所施加電壓為25 kV來進行市售番瀉錠劑定性與定量的工作。相較於一開始所使用的35 mM且pH值為9.0的硼酸鹽緩衝溶液，添加15% (v/v) ACN，溫度為30ºC，施加電壓為20 kV分離條件，此分析方法具備分析時間短，基線平穩，分離時所產生的電流小，且不需添加有機溶媒的優點，為市售的各種番瀉製劑提供了一快速、穩固的分析方法。

Part I: Analysis of anthraquinones and bianthrones in rhubarb by microemulsion electrokinetic chromatography Microemulsion eletrokinetic chromatograpy (MEEKC) is a separation technique which is similar in principle to micellar electrokinetic chromatography (MEKC), but with more parameters to be adjusted so as to obtain better separation results. The separation of nine anthraquinones and bianthrones in the present study was investigated for the key-operating variables such as the concentration and the type of surfactant, the type of oil phase, and the addition of organic solvent. The optimal condition was found to be 0.5% (w/w) di-n-butyl-L-tartrate, 1.2% (w/w) 1-butanol, 0.6% (w/w) SDS, 97.7% (w/w) 10 mM Na2B4O7 (pH 9.2) and ACN 30% (v/v), under which these nine compounds could be baseline separated within 20 minuntes. Owing to the complexity of rhubarb constituents, we used solid- phase extraction (SPE) for sample pretreatment. By utilizing suitable proportion of weak acid, weak base, and organic solvent for sample loading, washing, and eluting steps on an Oasis HLB sorbet, most of interference in the crude extracts of rhubarb could be cleaned off. The partially purified extracts with finely tuned concentrations could be injected into capillary electrophoresis. Thus, seven analytes were accurately quantitated. Part II: Analysis of sennoside A and sennoside B in commercial senna tablets by capillary zone electrophoresis Qualitative and quantitative analyses of sennoside A and sennoside B, the major cathartic constituents, in commercial senna tablets was performed in the present study by capillary zone electrophoresis. During the process of method development, we investigated the influence of possible parameters on resolution, including the type, concentration, and pH value of the buffer solution, the temperature, the applied voltage and the addition of organic solvent. The optimal condition was found to be 100 mM CAPS (pH 10.4) at 25 kV and 30