Laser microdissection hyphenated with high performance gel permeation chromatography-charged aerosol detector and ultra performance liquid chromatography-triple quadrupole mass spectrometry for histochemical analysis of polysaccharides in herbal medicine: Ginseng, a case study

Abstract

This study establishes a new combinatorial approach for histochemical analysis of polysaccharides in herbal medicines using laser microdissection followed by high performance gel permeation chromatography coupled with charged aerosol detector and ultra-performance liquid chromatography hyphenated with triple quadrupole mass spectrometry. Ginseng was employed as a study model. Tissue-specific qualitative and quantitative characterization of ginseng polysaccharides was performed by determining their molar masses and monosaccharide compositions in three macro-dissected parts (rhizome, main and branched roots) and five micro-dissected tissues (cork, cortex, xylem, phloem and resin canal). The results showed that ginseng “flesh” (xylem, phloem and resin canal) contained more polysaccharides with larger molecular weights and higher ratios of glucose residue, whereas ginseng “skin” (cork and cortex) had fewer polysaccharides with smaller molecular weights and higher ratios of non-glucose constituents (e.g. galacturonic acid, galactose, arabinose and rhamnose). These findings suggested that the polysaccharides of the “flesh” were predominantly starch-like glucans, while those of the “skin” were of a higher proportion of acidic pectins. The revealed histologic distribution and accumulation pattern of ginseng polysaccharides contributes to the scientific understanding of ginseng regarding the biosynthesis and transportation of polysaccharides, medicinal quality evaluation as well as empirical clinical application.