Application Of Two-dimensional Chromatography On The Separation Of Ingredient In Ginseng Saponins

With thousands of years of practice experience and history,traditional Chinese medicine has attracted attention from all of the word,However,the absent of understand of the material foundation set back its deep exploitation and utilization,As an powerful tool,Separation play a very important role in the study of the chemical constitutes of the Chinese herbs,how to establish high efficient,large-scaled and environment-protected purification methods are increasingly become the research focus to face the proposed of sustainable development and ecological civilization construction and "green chemistry".In this paper,Ginseng root extract rich in ginsenosides was chose as a research target and the research topic focus on the establishment of high efficient,large-scaled and environment-protected purification methods for ginsenosides separation and purification.This paper covers the following aspects:1.In order to solve the existing high solvent consumption,high pollution problems in traditional chromatography,We have designed the environment-friendly fast chromatographic system,with panaxdiol saponins crude extract as a separate object and have carried on a test to the system including chromatography time,elution volume,solvent loss,number of fractions,products separation effect,etc,proved that the system has good separation effect,low solvent volatilization and fast chromatography speed.Then using the system combined with hydrophilic interaction chromatography established a method based on normal phase silica gel column chromatography and hydrophilic interaction chromatography for the separation and purification of panaxdiol saponins.2?Froi the combination experience of traditional mass separation method and efficient separation method,we have chosen another separation materials macroporous resin which is suitable for large-scale application and combined with preparative liquid chromatography,aiming to establish a kind of scaled and more efficient isolation and purification method for saponin compounds.Serving ginseng root extract as a separate object and based on the molecular characteristics of ginseng saponin,we have established an off-line orthogonal two-dimensional separation methods HPD100 reversed-phase macroporous resin/preparative hydrophilic liquid chromatography.Separation mode selection and a range of HPLC gradient optimization,examined the method of sample processing capacity and separation efficiency and through a standard control,MS and MS2 characterizing compounds derived from preparation process.3.To address the application problems of comprehensive twodimensional liquid chromatography in preparative chromatography levels,we tried to establish a cycle-time controlled comprehensive two-dimensional liquid chromatography.By lifting the time limit in 2nd dimension separation,removing restrictions of injection volume,let it become possible that comprehensive two-dimensional chromatography amplifying to preparative level,improving the application of this method in separation and purification of natural products.The cycle time of this comprehensive two-dimensional chromatographic separation system can be controlled mainly by the stop-flow gradient,ten-way valve,double trap columns and 2%acetonitrile make-up flow(0.2 mL/min)-Ginseng root extract as a separate object,1st dimension and 2nd dimension using Spherigel C18 column and Click Xion column,respectively,and orthogonality of these two columns were investigated.Additionally,stop-flow gradient conditions on the 1st dimension,2nd dimensional hydrophilic interaction liquid chromatography conditions are optimized and characterized separation effect of the system.