目的 构建甘草黄酮类化合物系统性分离制备方法。方法 采用特异性吸附材料富集甘草中的黄酮类化合物，以自主研发的制备色谱工厂系统，采用色谱分离专家系统软件优化分离制备条件，通过上样量和富集次数等参数的考察，建立了基于分离富集模式的反相二维色谱制备甘草黄酮有效部位及单体化合物的方法。结果 建立了以C₁₈为分离、富集填料，甲醇-水、乙腈-水为一维、二维分离流动相，水为富集稀释液，梯度洗脱体积流量和稀释富集液的体积流量均为21 mL/min，上样量300 mg，富集次数3次的二维色谱分离制备甘草黄酮的方法，其分离过程具有良好的重复性。应用该方法分离制备，可重复获得16个甘草黄酮部位和甘草苷、甘草素、芒柄花黄素、刺甘草查耳酮、7，4'-二羟基黄酮、4'-O-[β-D-apio-D-furanosyl-（1→2）-β-D-glucopyranosyl]liquiritigenin、异甘草素、甘草酚、甘草香豆素共9个单体化合物。结论 建立的甘草黄酮的制备方法为甘草资源的综合利用和甘草黄酮活性药物开发奠定了基础。

Objective To develop a systematic chromatography separation method for flavonoids from Glycyrrhiza uralensis Fisch.(GU).Methods A new method for the separation of effective parts and monomers of flavonoids from GU by two-dimensional reversed-phase liquid chromatography was developed using the self-developed preparation chromatography plant system with independent intellectual property rights.Flavonoids compounds were enriched with specific adsorption materials.The separation conditions of the chromatography were optimized by the chromatographic separation expert system software, and the loading weight of samples and the enrichment times of a separation were investigated.Results The process of chromatography separation of flavonoids from GU had good precision and reproducibility with C₁₈ 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 gradient elution and dilution enrichment solution was 21 mL/min.The sample loading amount of chromatography separation was 300 mg each time.A total of 16 flavonoids parts contained stable chemical composition were obtained by the repeatable separation method after three times of enrichment.Nine pure compounds were obtained and identified by NMR and MS, which were liquiritin, liquiritigenin, formononetin, echinatin, 7, 4'-dihydroxyflavone, 4'-O-[β-D-apio-D-furanosyl-(1→2)-β-D-glucopyranosyl]liquiritigenin, isoliquiritigenin, glycyrol, and glycycoumarin.Conclusion The study can provide a certain reference value for the systematic separation and cognition for flavonoids from GU.

国家自然科学基金-联合基金项目（U1603285）；辽宁省教育厅基本科研项目（2016J008）；江苏省“双创计划”项目（苏人才[2015]26号）；连云港市“高层次创新创业领军人才集聚工程”资助（连人才[2016]1号）