Study On Separation And Enrichment Of Lignans From Schisandra Chinensis

The chemical composition of traditional Chinese medicine was very complex and the content of bioactive sites was less.Therefore,the separation and purification of the chemical composition of traditional Chinese medicinewere important.Chromatography was the core technology to separate and purify traditional Chinese medicine.At present,the systematic separation of chemical components of Chinese medicinestill could not be solved.Therefore,in this paper,schisandra chinensis was used as the research object.Preparative Chromatography Platform combined with the expert system software to develop a systematic and high-throughput separation method.This Preparative Chromatography Platform was a two-dimensional reversed phase liquid chromatography and based on the modules of separation and enrichment.It was self-developed with independent intellectual property rights.It also provides reference for separation and preparation of other Chinese medicinal ingredients.In this paper,the crude extract of Schisandra chinensis was used as the object of study.TLC and HPLC was used as the testing method.The extraction solvent and the proportion of elution are studied,and the lignans of Schisandra chinensis were preliminarily separated.The results showed that extraction was a simple and effective pre-treatment technology.Ethyl acetate and petroleum ether were selected as solvent system（1:1）.which could preliminarily Separate and enrichment lignans from Schisandra chinensis.The extraction sample of Schisandra chinensis is the object of study,HPLC was used as test facility.The chromatographic column was selected as C18 column（250×4.6mm;5μm）,andwater and methanol was selected as mobile phase.30 chromatographic peaks were selected and they information of 4 times gradient chromatography of the lignans extracts were applied to the chromatographic separation expert system software-XTool.The separation conditions of the first and second dimensional chromatographic separation were optimized by XTool.The results show that the retention time of the chromatographic peak of Schisandra chinensis was in accordance with the retention time of chromatographic peaks under gradient elution conditions simulated by xtool software.It was proved that the method of the chromatographic separation expert system to predict elution conditionsis feasible.Then,a linear amplification was used for preparative chromatography.As a result,the Sample concentration was 80mg/mL,the sample size was 2mL,and the flow rate was21ml/min.At this time the chromatographic peak has obvious separation.The column C₁₈（250mm×30 mm,10μm）was selected as first dimensional separation column.Water and methanol were used as mobile phases.The columns C₁₈（80 mm×30 mm,10μm）were selected as enriching columns and water as the diluent solution.Chromatographic retention timewas selected as investigation index,The relative standard deviations of chromatographic retention time after first dimensional separation of 3 times were all less than 3%.It shows that the reversed-phase two-dimensional chromatographic separation has good reproducibility.Finally,9repeatable lignanscomponents were obtained by first dimensional separation.PCP was swithched to second dimensional separation mode.The column C₁₈（250 mm×30 mm,10μm）was selected as second dimensional separation column.The sample size,flow rate and mobile phases were the same as first dimension separation.20 repeatable lignanscomponents were obtained by second dimensional separation.Under one dimensional separation mode of Preparative Chromatography Platform,10two-dimensional separation samples with purity of 90%were separated and purified by Preparative Chromatography Platform.5 monomers compounds including Schisandrol A、Gomisin A、Schizandrin A、Gomisin N、Schisandrin C were identified by TLC,HPLC,¹H-NMR,¹³C-NMR spectroscopy testing methods.Experimental results show that The chromatographic expert system software combined with the Preparative Chromatography Platform to simulate and optimize the preparation conditions for the separation of Schisandra lignans.A new method for the separation and preparation of the effective parts and monomers of Schisandra lignans was established.This method has good reproducibility and could achieve systematic,high throughput and high purity separation of lignans from Schisandra chinensis.It is significant for the separation of lignans from Schisandra chinensis and the preparation of other traditional Chinese medicine components.