人参中皂苷类化合物的HPLC-ESI MS研究

利用高效液相色谱与电喷雾质谱联用技术分析鉴定了人参粗提物中人参皂苷类化合物。实验采用反相C18色谱柱，二元线性梯度洗脱（0．2％的醋酸水溶液和乙腈），分离并检测了人参中的皂苷类化合物：通过与电喷雾质谱的联用和质谱的源内CID技术获得了其中主要色谱峰相对应化合物的相对分子质量和结构信息。     

源内裂解液相色谱-质谱鉴别人参皂甙砒和拟人参皂甙F11

研究了利用源内碰撞诱导解离（in-source collision—induced dissociation）的高效液相色谱-大气压化学电离质谱（HPLC—APCI／MS）获取人参和西洋参的化学标志物——人参皂甙Rf和拟人参皂甙F11的特征结构信息及鉴别人参和西洋参的方法。在乙腈-水梯度洗脱反相液相色谱及源内碰撞诱导解离条件下，能获得人参皂甙Rf和拟人参皂甙F11的母核离子及去糖基离子的源内碰撞诱导解离谱，从其差别能清楚区分这对同分异构体。本方法对人参皂甙Rf和拟人参皂甙F11的检出限能达到10^-7g柱上样量，简单、快速，单次质谱实验就能鉴别人参和西洋参。     

七叶一枝花中薯蓣皂苷的分离及结构鉴定研究

本文利用高效液相色谱-串联质谱联用方法研究了七叶一枝花中的薯蓣皂苷。实验采用高效液相色谱分离了七叶一枝花中的3种薯蓣皂苷;通过与电喷雾质谱联用获得了这几种化合物的分子量信息;再用MS/MS获得了这几种化合物进一步的结构信息。采用此方法可快速分析鉴定从七叶一枝花中分离得到的薯蓣皂苷。     

应用高效液相色谱和电喷雾质谱联用技术分析鉴定车前草中的苯乙醇苷化合物

本文采用高效液相色谱与电喷雾质谱联用技术在线分析鉴定了车前草提取物中的三种苯乙醇苷化合物。实验采用反相C18色谱柱,0.2%的醋酸水溶液和乙腈梯度洗脱,车前草中的苯乙醇苷化合物得到很好的分离。在电喷雾质谱负离子条件下,获得了三种苯乙醇苷化合物的分子离子峰,分子量信息,进一步通过质谱的源内CID技术得到相应化合物的结构信息。通过得到的这些信息与文献中的已知化合物或标准品对照从而推断出化合物的结构。     

超高效液相色谱-四极杆静电场轨道阱高分辨质谱联用法分析不同西洋参加工品的皂苷类成分

采用超高效液相色谱-四极杆静电场轨道阱高分辨质谱联用法(UPLC Orbitrap HRMS)对西洋参不同加工品中的皂苷类成分差异进行分析。高分辨质谱条件为电喷雾电离源,轨道阱质量分析器,负离子模式采集。结果显示,根据一级质谱精确质荷比信息,获得化合物的精确分子量;根据二级串联质谱信息,获得化合物的碎片信息。总结了不同类型皂苷的特征质谱碎裂规律,并对碎裂规律进行比对,初步归属和鉴定西洋参中55种皂苷类成分,其中原人参二醇型皂苷成分28种、原人参三醇型皂苷成分17种、奥克梯隆型皂苷成分5种和齐墩果烷型皂苷成分5种。通过比较不同西洋参加工品中皂苷类组成的差异,为揭示不同加工方法对西洋参皂苷成分的影响规律提供了依据,并为西洋参的开发利用以及阐明药效物质基础提供了参考。     

高效液相色谱-电喷雾飞行时间质谱分析娑罗子中皂苷类成分

采用高效液相色谱/电喷雾飞行时间质谱联用技术(HPLC/ESI-TOF/MS),研究4种七叶皂苷的分子结构与裂解规律间的关系,并对娑罗子中的七叶皂苷类化合物进行鉴定。实验采用反相C18色谱柱,以乙腈-0.2%乙酸溶液为流动相,二元线性梯度洗脱,通过与电喷雾飞行时间质谱联用获得娑罗子中各皂苷成分的精确分子量和分子式;采用质谱碰撞诱导解离技术获得各化合物碎片裂解信息,结合文献对娑罗子中的14种皂苷类化合物进行了初步鉴定。研究表明,高效液相色谱-电喷雾飞行时间质谱联用技术是娑罗子中皂苷类化合物鉴别的有效工具。     

源内裂解液相色谱-质谱鉴别人参皂甙Rf和拟人参皂甙F11

研究了利用源内碰撞诱导解离(in-source collision-induced d issoc iation)的高效液相色谱-大气压化学电离质谱(HPLC-APC I/MS)获取人参和西洋参的化学标志物———人参皂甙R f和拟人参皂甙F11的特征结构信息及鉴别人参和西洋参的方法。在乙腈-水梯度洗脱反相液相色谱及源内碰撞诱导解离条件下,能获得人参皂甙R f和拟人参皂甙F11的母核离子及去糖基离子的源内碰撞诱导解离谱,从其差别能清楚区分这对同分异构体。本方法对人参皂甙R f和拟人参皂甙F11的检出限能达到10-7g柱上样量,简单、快速,单次质谱实验就能鉴别人参和西洋参。     

朝鲜淫羊藿中黄酮类化合物的高效液相色谱与电喷雾质谱联用研究

利用高效液相色谱与电喷雾质谱联用技术研究了朝鲜淫羊藿中的黄酮类化合物。实验采用反相C18色谱柱，二元线性梯度洗脱，分离并检测了朝鲜淫羊藿中的6种黄酮类化合物；通过与电喷雾质谱联用获得了这几种化合物的分子量信息，利用质谱的源内碰撞诱导解离技术对这几种化合物进行了结构鉴定。     

聚类分析法用于西洋参样品分类研究

建立西洋参药材分类方法；分别采用电感耦合等离子体质谱（ICP—MS）法、高效液相色谱（HPLC）法以及蒽酮-硫酸分光光度法，对12个西洋参样品中的15种无机元素、7种人参皂苷和多糖进行测定，用SPSS聚类分析法对12个西洋参样品进行聚类分析；结果表明：以4种不同的聚类变量进行聚类分析，其聚类分组结果并不一致，将几种有效成分综合为聚类变量，对西洋参进行分类更为科学合理；聚类分析法是西洋参分类分析的有效方法，对西洋参品种与质量的鉴定有一定的理论意义和实用价值。     

纳升电喷雾萃取电离质谱快速测定人参皂苷

以自行研制的纳升电喷雾萃取电离源（NanoEESI）为基础,建立了在无需色谱分离条件下快速测定人参皂苷的质谱分析方法,用NanoEESI串联质谱分析鉴定了人参中的多种人参皂苷,并将该方法用于人参、桔梗和商陆的快速鉴定。实验表明,人参中的皂苷易于在电离过程中结合钠离子,并形成[M+Na]+正离子从而被检测分析,而桔梗和商陆等样品中不含人参皂苷成份,在串联质谱中也没有相应的特征碎片离子,从而达到鉴别中药材真伪的目的。本方法简便、快速、灵敏、特征性强、重现性好,对道地中药材快速鉴定提供了思路,具有很好的实用价值。     

Determination of ginsenosides (ginseng saponins) in dry root powder from Panax ginseng, Panax quinquefolius, and selected commercial products by liquid chromatography: interlaboratory study

Twelve collaborating laboratories assayed 4 products, namely, Panax ginseng, Panax quinquefolius, and 2 ginseng products, for 6 ginsenosides: Rb1, Rb2, Rc, Rd, Re, and Rg1. Collaborators also received a negative control for the recovery study. Pure ginsenosides were provided as reference standards for the liquid chromatography (LC) analysis and the system suitability tests. The LC analyses were performed on the methanol extract using UV detection at 203 nm. For P. ginseng, individual ginsenosides were consistent in their means; repeatability standard deviations (RSDr) ranged from 4.17 to 5.09% and reproducibility standard deviations (RSDR) ranged from 7.27 to 11.3%. For P. quinquefolius, the Rb1 and Rb2 ginsenosides were higher and lower in concentration than P. ginseng, with RSDr values of 3.44 and 6.60% and RSDR values of 5.91 and 12.6% respectively, and other analytes at intermediate precisions. For ginseng commercial products, RSDr values ranged from 3.39 to 8.12%, and RSDR values ranged from 7.65 to 16.5%. A recovery study was also conducted for 3 ginsenosides: Rg1, Re, and Rb1. The average recoveries were 99.9, 96.2, and 92.3%, respectively. The method is not applicable for the determination of Rg1 and Re in ginseng product at levels <300 mg/kg.     

Determination of ginsenosides in Panax ginseng roots by liquid chromatography with evaporative light-scattering detection

A liquid chromatographic method was developed and validated for the determination of ginsenosides in Panax ginseng roots by using evaporative light-scattering detection. Eighteen ginsenosides were separated on a reversed-phase C18 column with water-ammonium acetate-acetonitrile as the mobile phase. The method is suitable for the routine determination of ginsenosides in P. ginseng roots and extracts. The validation of the method was comprehensive for efficiency and recovery optimization of the P. ginseng roots extraction, specificity by liquid chromatography/mass spectrometry, linearity, stability, reproducibility, repeatability, intermediate precision, and robustness.     

Development of a rapid and convenient method to separate eight ginsenosides from Panax ginseng by high-speed counter-current chromatography coupled with evaporative light scattering detection

Ginsenosides exhibit diverse biological activities and are major well-known components isolated from the radix of Panax ginseng C.A. Meyer. In the present work, a rapid and facile method for the separation and purification of eight ginsenosides from P. ginseng by high-speed counter-current chromatography coupled with evaporative light scattering detector (HSCCC-ELSD) was successfully developed. The crude samples for HSCCC separation were first purified from ginseng extract using a macroporous resin; the extract was loaded onto a Diaion-HP20 column and fractionated by methanol and water gradient elution. The ginsenosides-protopanaxadiol (PPD) and protopanaxatriol (PPT) fractions were subsequently eluted with 65 and 80% methanol and water gradient elution, respectively. Furthermore, these two fractions were separated by HSCCC-ELSD. The two-phase solvent system used for separation was composed of chloroform/methanol/water/isopropanol at a volume ratio of 4:3:2:1. Each fraction obtained was collected and dried, yielding the following eight ginsenosides: Rg(1), Re, Rf, Rh(1), Rb(1), Rc Rb(2) and Rd. The purity of these ginsenosides was greater than 97% as assessed by HPLC-ELSD, and their structures were characterized by electrospray-ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance spectroscopy. This is the first report regarding the separation of the ginsenosides Rh(1), Rb(2) and Rc from P. ginseng by HSCCC.     

Determination of polyacetylenes and ginsenosides in Panax species using high performance liquid chromatography

A new HPLC method was developed to separate and identify three polyacetylenes (panaxynol, panaxydol and 1,8-heptadecadiene-4,6-diyne-3,10-diol) found in Panax species. The mobile phase was a linear gradient of 2 : 1 : 3 to 2 : 1 : 1 (v/v/v) methanol/acetonitrile/water in 40 min. HPLC analysis was performed at a flow rate of 1.5 ml/min with UV detection at 254 nm. The contents of the polyacetylenes and ginsenosides in Panax ginseng (white ginseng and red ginseng), P. quinquefolium, P. japonicus, and P. noteginseng were determined using these methods. The species containing the highest polyacetylene content (0.080%) was P. quinquefolium cultivated in Nagano, Japan. Meanwhile, the species with the highest ginsenoside content (9.176%) was P. noteginseng cultivated in Yunnan, China.     

Analysis of Low-polar Ginsenosides in Steamed Panax Ginseng at High-temperature by HPLC-ESI-MS/MS

A high performance liquid chromatography coupled with electrospray ionization-tandem mass spectrometry( HPLC-ESI-MS/MS) method was developed for the analysis and identification of ginsenosides in the extracts of raw Panax ginseng(RPG) and steamed Panax ginseng at high temperatures(SPGHT). A total of 25 ginsenosides were extracted include of which 10 low-polar ginsenosides, such as ginsenosides F4, Rk3, Rh4, 20S-Rg3, 20R-Rg3 and so on, were identified according to their HPLC retention time and MS/MS data. The results indicated that the low polar ginsenosides were seldom found in RPG. For the exploration of the transformation pattern of the ginsenosides in steam processing, the standards of ginsenosides Re, Rg1, Rb1, Rc, Rb2, Rb3 and Rd were selected and hydrolyzed at a temperature of 120 ℃. The results show that these polar ginsenosides can be converted to low-polar ginsenosides such as Rg2, Rg6, F4, Rk3 and Rg5 by hydrolyzing the sugar chains.     

人参中人参皂苷的直接高压微波辅助降解

采用高效液相色谱-电喷雾质谱联用法测定了人参提取液中的人参皂苷. 考察了天然人参皂苷发生降解的条件, 同时研究了单体人参皂苷Rg1, Re, Rb1, Rc, Rb2和Rd的降解, 并对降解产物进行了分析. 结果表明, 随着提取压力的升高, 提取液中天然人参皂苷的含量逐渐减少, 同时产生多种次级人参皂苷. 当微波提取压力达到600 kPa, 提取时间为10 min时, 提取液中的主要天然人参皂苷达到完全降解, 次级人参皂苷Rg3含量达到最高. 在单体人参皂苷Rb1, Rc, Rb2和Rd的降解产物中均得到人参皂苷Rg3.     

Determination of Saponins in Leaf of Panax Ginseng C. A. Mey. by High Performance Liquid Chromatography

A high performance liquid chromatography(HPLC) with UV detection was established for simultaneous determination of saponins in the leaf of Panax ginseng C. A. Mey. Nine ginsenosides(Rb1,Rb2,Rb3,Rc,Rd,F1,F2,F3,F5) and notoginsenoside Fe(NFe) were studied. Among the saponins,the ginsenosides F1,F2,F3,F5 and NFe were determined by HPLC-UV method for the first time. The determination of the ginsenosides via the HPLC-UV method was performed on a reversed-phase C18 column with gradient elution in 40 min. The line...     

Identification of Ginsenosides in Panax quinquefolium by LC-MS

An HPLC-APCI-MS method for the identification of ginsenosides in Panax quinquefolium has been developed. HPLC-APCI-MS could effectively identify ocotillol, protopanaxadiol, protopanaxatriol and oleanane-type ginsenosides in a single MS experiment since [M-H]⁻ ions and characteristic thermal degradation ions of ginsenosides could be simultaneously observed under negative and positive ionization conditions. Nine ocotillol-type ginsenosides including 24(R)-pseudoginsenoside F₁₁ were firstly identified and a total of 30 ginsenosides were identified in Panax quinquefolium. The ginsenoside profile differences between Chinese and American P. quinquefolium were investigated by HPLC-APCI-MS.     

Preparative isolation of four ginsenosides from Korean red ginseng (steam-treated Panax ginseng C. A. Meyer), by high-speed counter-current chromatography coupled with evaporative light scattering detection

Ginseng (Panax ginseng C. A. Meyer) has been well known to have a variety of ginsenosides that show diverse biological activities. Especially, the components of ginsenosides are quite different depending on the processing method. Recently, there have been several reports showing that less polar ginsenosides from Korean red ginseng (steam-treated Panax ginseng) have potent biological activities such as radical scavenging, vasodilating and anti-tumor activities. In this study, we have isolated four known ginsenosides Rg3, Rk1, Rg5 and F4 from Korean red ginseng by high-speed counter-current chromatography (HSCCC) coupled with evaporative light scattering detection (ELSD). The enriched saponin fraction (350 mg) was separated by using methylene chloride-methanol-water-isopropanol (6:6:4:1, v/v) as the two-phase solvent system and yielded 28.6 mg of Rg5, 26.6 mg of Rk1, 32.2 mg of Rg3 and 8.1 mg of F4. The purity of these ginsenosides was assessed by HPLC-ELSD to be over 95%, and their structures were characterized by electrospray ionization mass spectrometry (ESI-MS), (1)H NMR and (13)C NMR.