台灣蚤休中類固醇皂苷的分離與分析

Isolation and analysis of steroidal saponins in Taiwanese Rhizoma Paridis curtivar (Paris formosana HAYATA)

呂美鵬(Mei-Peng Lu)

指導教授 ： 林昭田

中山醫學大學/健康管理學院/應用化學系碩士班/碩士(2010年)

https://doi.org/10.6834/CSMU.2010.00062

摘要

蚤休為百合科（Liliaceae）重樓屬（Paris genus）多年生草本植物,其中含的固醇類皂苷(steroidal saponins)為其重要的生理活性化合物。由於台灣產蚤休植物中的皂苷組成與含量仍不清楚，有必要加以釐清。本實驗廣泛收集各地台灣蚤休（Paris formosana HAYATA），並由其分離純化類固醇皂苷，再進行結構鑑定，以確認台灣蚤休中的類固醇皂苷組成。並以高效能液相層析法（high performance liquid chromatography , HPLC）配合蒸發光散射檢測系統（evaporative light scattering detector, ELSD），建立一套檢測蚤休類固醇皂苷含量的分析方法，再分析台灣各產地蚤休中類固醇皂苷組成的差異，確立其中類固醇皂苷的含量。本實驗以甲醇進行蚤休皂苷粗萃,接著以正丁醇進行液-液萃取（liquid-liquid partition），最後以HPLC分離出9個類固醇皂苷，經液相層析質譜儀（LC-MS）和核磁共振儀（NMR）進行結構鑑定，發現其分別為Methyldichotomin、PolyPhyllin H、Methylprotogracillin、Paris-VII、Chonglouoside H、Formosanin C、Gracillin、Polyphyllin D和Paris V。所建立之分析方法是以C18為固定相（stationary phase）氰甲烷-水為移動相（mobile phase）（前30分鐘比例為: 25/75(v/v), 31~45分鐘為: 45/55(v/v),45~65分鐘為: 50/50(v/v),流速為1 ml/min），可同步分離這些類固醇皂苷。檢測各產地蚤休中類固醇皂苷的含量時，發現南投與台東地區的蚤休皂苷含量與種類較多。

關鍵字

台灣蚤休 ； 類固醇皂苷純化 ； 分析

並列摘要

Rhizoma Paridis is a perennial herbaceous plant of the Paris genus (belong to Liliaceae family). Steroidal saponins are the notable bioactive compounds of the herb. However, there is no thorough report concerning composition and content of steroidal saponins in Taiwanese Rhizoma Paridis plants. In the study, we collected Paris formosana HAYATA from various areas of Taiwan followed by isolation, purification and identification of steroidal saponins. A simple method using HPLC-ELSD (evaporative light scattering detection, ELSD) for simultaneous determination of these steroidal saponins was also developed. The method was applied to determine the composition and content of steroidal saponins in the herb samples.
　　Dried methanol extract of P. formosana HAYATA was suspended in water and then partitioned against by n-butanol to yield saponin extract. The extract could be isolated nine steroidal saponins by high preformance liquid chromatography (HPLC). Their structures were identified by LC-MS and NMR. They are three furostanol glycosides including Methyldichotomin, PolyPhyllin H and Methylprotogracillin, and two pennogenin glycosides (spirostanol glycosides) including Paris-VII and Chonglouoside H, and four diosgenin glycosides (spirostanol glycosides) including Formosanin C, Gracillin, Polyphyllin D and Paris V. The established analytical conditions used to determine these saponins were a C18 column (35°C) and a step gradient solvent system consisting of acetonitrile and deionized water （25/75(v/v) in the first 30 min, 45/55(v/v) from 31 to 45 min and 50/50 (v/v) from 45 to 65 min） at a flow rate of 1 ml/min. The P. formosana HAYATA gathered from Nantou and Taitung had higher levels of steroidal saponins.

並列關鍵字

Taiwanese Rhizoma Paridis curtivar (Paris formosana HAYATA) ； steroidal saponins

參考文獻

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