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J Pharm Biomed Anal. 2012 Feb 5;59:67-77. doi: 10.1016/j.jpba.2011.10.004. Epub 2011 Oct 13.

Target separation of a new anti-tumor saponin and metabolic profiling of leaves of Panax notoginseng by liquid chromatography with eletrospray ionization quadrupole time-of-flight mass spectrometry.

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State Laboratory of Modern Chinese Medicines, China Pharmaceutical University, Nanjing 210009, PR China.


A method coupling high-performance liquid chromatography (HPLC) with quadrupole time-of-flight mass spectrometers (QTOF-MS) using an eletrospray ionization (ESI) source was firstly developed for detection, characterization and guiding target separation of variants of protopanaxdiol saponin from leaves of Panax notoginseng. Under the guidance of LC-QTOF-MS, a new trace saponin was probed according to the precise elemental compositions of molecular ions and the fragmentation behavior, and then separated from the ethanol extract of the plant by a set of chromatographic methods. It was further confirmed by NMR experiments as 3-O-β-D-glucopyranoside-3β,l2β,23β-triol-20-ene-dammar (Pn-1). The cytotoxic assay showed that Pn-1 had relatively stronger anti-tumor effects against three tumor cell lines (NCI-H460, HepG2 and SGC-7901) than Rg₃, an approved clinical agent for cancer therapy. Meanwhile, based on accurate mass measurements within 5 ppm for each molecular ions and subsequent product ions, 48 saponins, including 40 protopanaxadiol saponins, 7 protopanaxatriol saponins and 1 oleanane saponin were identified. It is noted that the knowledge of the presence of abundant protopanaxadiol saponins in leaves of P. notoginseng may provide tools for a full understanding of the chemical diversity of secondary metabolites from the different parts of P. notoginseng. From the points of time consuming and accurate mass measurement capability, the LC-QTOF-MS is a highly powerful tool for screening and guiding target separation of new compounds in herbal extract, and thus benefits the speed of new drug discovery progress.

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