利用液相層析串聯高解析質譜法建立人參皂苷及皂苷元分析平台

Abstract

人參皂苷 (ginsenosides) 為人參 (ginseng) 中主要生理活性成分。為了因應人參原料及其萃取物品質管制之需求，本研究使用液相層析串聯高解析度軌道阱質譜儀 (ultra-high performance liquid chromatography coupled to high resolution (Orbitrap) mass spectrometry, LC-HRMS/MS) 建立人參皂苷及其相關化合物化學指紋圖 (chemical fingerprint profile) 之分析平台。此分析平台可辨識出62種人參皂苷及4種皂苷元 (sapogenins) ，其中包含17種原人參三醇 (protopanaxatriol, PPT)、34種原人參二醇 (protopanaxadiol, PPD) 、4種奧克梯隆型 (ocotillol type, OT)、4種齊墩果烷酸型 (oleanolic acid type, OA) 及7種C-17分支多變型 (C17 side-chain varied type) 人參皂苷。分析平台亦可辨識6種乙醯化 (acetyl-) 、8種丙二醯化 (malonyl-)、1種丁醯化 (butenoyl-) 及1種辛醯化 (octenoyl-) 人參皂苷。研究結果顯示亞洲參根、西洋參根及三七根皆未含有皂苷元(苷元)；各品種人參之人參皂苷以PPT型及PPD型人參皂苷為大宗，而OT型、OA型人參皂苷和C-17分支多變型人參皂苷則是次要成分。針對可取得的樣品發現西洋參之次要人參皂苷成分相較於亞洲參和三七來得豐富。本研究亦發現紅參中低極性人參皂苷含量落在0.0875 至0.2708 mg/g範圍顯著高於新鮮人參，顯示出加工製備所產之紅參在熱處理下的變化。根據人參皂苷之組成，藉由主成分分析 (principal components analysis) 區分亞洲參、西洋參及三七。本研究利用此分析平台可有效辨識人參皂苷及皂苷元指紋圖譜及應用於人參原料和相關產品進行品質管制及加工監控指標之用途。

Ginsenosides are the major bioactive ingredients in ginseng. By responding to the demanding for quality control of the raw materials and extracts of ginseng, this study established a platform for chemical fingerprint profile of ginsenosides and their related compounds on an ultra-high performance liquid chromatography coupled to high resolution (Orbitrap) mass spectrometry (LC-HRMS/MS). The platform can detect 62 ginsenosides and 4 sapogenins, including 17 protopanaxatriol-type (PPT), 34 protopanaxadiol-type (PPD), 4 ocotillol-type (OT), 4 oleanolic acid-type (OA) and 7 C17 side chain varied-type ginsenosides. There were also 6 acetyl-ginsenosides, 8 malonyl-ginsenosides, 1 butenoyl- ginsenosides, 1 octenoyl-ginsenosides that can be identified. We did not find any sapogenin, aglycone, in selected raw materials, including radix of Panax ginseng, Panax quinquefolius, and Panax notoginseng. The PPT- and PPD-type components were major ginsenosides in Panax species with less content of OT-type, OA-type, and C17 side chain varied-type ginsenosides. For comparison of those minor constituents in the selected samples, P. quinquefolius were higher than those in P. ginseng and P. notoginseng. The content of low polarity ginsenosides in red ginsengs was in the range of 0.0875 to 0.2708 mg/g, which were significantly higher than fresh ginsengs. It could be the result of the thermal treatment of the red ginseng preparing process. According to their ginsenoside composition, the statistical principal component analysis can differentiate P. quinquefolius, P. ginseng, and P. notoginseng. The analytical platform established can be an effective tool to obtain the fingerprint of ginsenosides and sapogenins for quality control of ginseng raw material and products and indices of process monitoring.