| Platycodon grandiflorum,the root of Platycodon grandiflorum A.DC?Campanulaceae?,is a typical traditional Chinese medicine cultivated all over the country used as both medicine and food.Studies show that Platycodon grandiflorum mainly contains triterpenoid saponins,fatty acids,flavonoids,steroids,polysaccharides,phenolic acids,polyacetylenes,and other types of compounds.There are more reports on triterpenoid saponins but relatively few studies on polyacetylene in Platycodon grandiflorum.However,the content of polyacetylenes like lobetyolin is high and it has many pharmacological activities,which has great research value.Three polyacetylenes were isolated and purified from Platycodon grandiflorum by resin-based column chromatography,High-speed counter-current chromatography,and analytical reversed-phase HPLC.Their chemical structures were evaluated by high-resolution electrospray ionization time-of-flight mass spectrometry and one-and two-dimensional nuclear magnetic resonance spectroscopy analyses,proving they are platetyolin A,platetyolin B and lobetyolin,among which,platetyolin A and platetyolin B are newly reported compounds.The methodology used in the present work for the isolation and purification of these polyacetylenes was suitable to obtain the compounds in sufficient amounts and a satisfactory degree of purity to allow further bioactivity studies to be performed with them.15 batches of Platycodon grandiflorum from different habitats was collected,and a HPLC method was established to determine the content of three kinds of polyacetylenes,It was found that the contents of three polyacetylene compounds in Platycodon grandiflorum from different habitats varied greatly,among which the total contents of the three polyacetylene compounds in Weifang,Shandong Province were the highest,which was 470.3?g·g–1.The established method is simple,accurate and reproducible,which lays the foundation for the application of these three polyacetylenes as a new quality control index in Platycodon grandiflorum.An UPLC method for simultaneous determination of 13 nucleosides in Platycodon grandiflorum collected from different habitats was also established,which found the highest content of nucleosides was1451.40?g·g–1 in Shandong,and the lowest was 325.86?g·g–1 in Nanyang.The established method can provide a new reference for the quality control and application of Platycodon grandiflorum.Gentianae radix is the root and rhizome of Gentiana manshurica Kitag.,Gentiana scabra Bge.,Gentiana rigescens Franch.,and Gentiana triflora Pal1.,its main chemical components are gentiopicroside,swertiamarin and other iridoids,which has various pharmacological activities.In this paper,four iridoids,loganic acid,swertiamarin,gentiopicroside and trifloroside were isolated and purified from Gentianae radix by high speed countercurrent chromatography.Their inhibition of NO release induced by LPS were studied.It was found that all four iridoids could inhibit the production of NO in BV-2 microglia induced by lipopolysaccharide in a dose-dependent manner,the inhibition rate of NO was the highest at 250?M of loganic acid,swertiamarin,gentiopicroside and trifloroside with the values of 58.1±10.4%,40.8±1.3%,73.3±1.1%,and 41.0±1.3%,respectively,and these four iridoids had no effect on cell activity,indicating that they could be used as an effective NO antagonist.In this paper,the Box-Behnken design was used to optimize experimental conditions for the pressurized liquid extraction of three iridoids?swertiamarin,gentiopicroside,and trifloroside?from Gentianae Radix.The optimum extraction conditions were as follows:ethanol concentration 61%,temperature 102°C,extraction time 18 min,diatomite addition amount 5.0 g.Under this condition,the extraction rate of iridoids were 72.02±1.06 mg·g–1.Consistently high yields in small extraction times were achieved with optimized pressurized liquid extraction,as compared to ultrasound-assisted and soxhlet extractions,pressurized liquid extraction is suitable for the extraction of iridoids from Gentianae Radix. |
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