Study On The Separation Of Flavonoids From Glycyrrhiza Uralensis Fisch By Two Dimensional Reversed-Phase Liquid Chromatography Of Separation-Enrichment Model

The compounds in traditional Chinese herb were complicated.It has a great significance to study active components in Chinese herb and create a new type of Chinese medicine through systematic separate of chemicalcomponents in Chinese herbs.To develop an efficient,high-throughput,systematic separation of Chinese medicinal ingredients was a prerequisite for the study of the basis of the pharmacodynamic substances of Chinese medicine.It was difficulttoachievehighthroughputandsystematicseparationbytraditional chromatographic separation.In the field of Chinese native medicine ingredient preparation,professional two-dimensional preparative chromatography separation of the development of Chinese medicine effective component and device has not yet been reported,and the conventional liquid chromatography in the separation of sample quantity was little,get effective parts of quality and poor repeatability,systemic and online operation means less complex,which has restricted the effective ingredients of traditional Chinese medicine systemic cognition and the active ingredient research.Separation strategy based on"the traditional Chinese medicine（TCM）chemical composition system and the equipment research",with licorice flavonoids richment content for the object,n new method on separation of flavonoids from Glycyrrhiza uralensis Fisch by two dimensional reversed-phase liquid chromatography was developed using the preparation chromatography plant system which self-developed with independent intellectual property rights.The separation conditions of the chromatography optimized by the chromatographic separation expert system software.The loading weight of a chromatography and the enrichment times of a separation were inspected.Contents and results are as follows:1.Extraction and enrichment of licorice flavonoids.The ural licorice root as raw material,after crushing soak extract,licorice flavonoid extract,enriched by polyamide chromatography packing,and then through LH20 glucangel chromatography packing processing enrichment of licorice flavonoids content of 17.2 g,47.8%flavonoids content.2.The process of chromatography separation of flavonoids from licorice had good precision and reproducibility with C₁₈8 as separation and enrichment solid phase and the methanol/water and acetonitrile/water as mobile phase of one-dimensional and two-dimensional chromatography system.The dilution solution which used for one-dimensional and two-dimensional enrichment chromatography was water.The flow rate of mobile phase and dilution solution and were 21 mL/min.The sample loading amount of one-dimensional chromatography separation was 300 mg each times.Three times one-dimensional chromatography separation were enrichment to one enrichment column.3.In this study,the method was used to prepare one dimension and two dimensional separation.The obtained monomer compounds were identified.Eventually,get 9 individual compounds,the structure was identified as liquiritigenin,formononetin,echinatin,7,4’-dihydroxyflavone,4’-O-[β-D-apio-D-furanosyl-（1→2）-β-D-glucopyranosyl]liquiritigenin,isoliquiritigenin,glycyrol,glycycoumarin.