title: Purification of high-purity glycyrrhizin from licorice using hydrophilic interaction solid phase extraction coupled with preparative reversed-phase liquid chromatography.
abstract: Glycyrrhizin (GA), a major bioactive compound in licorice, has been extensively used throughout the world as a medicine to treat chronic viral hepatitis and allergic dermatitis. In this study, a new method based on hydrophilic interaction solid phase extraction (HILIC-SPE) and preparative reversed-phase liquid chromatography (prep-RPLC) was developed to purify GA with high purity from the complex licorice extract. Via evaluation of retention behavior of GA and flavonoids in different commercially available columns, a hydrophilic column--Click XIon was finally chosen for the purification due to its excellent resolution toward GA and flavonoids under HILIC mode. To optimize the SPE elution conditions, relative factors including water content, pH and ionic strength had been investigated in chromatographic condition. The result indicated that the most appropriate water content was 30% and pH at 4.00, as well as salt concentration should be controlled at 5mM. In addition, the optimization revealed that GA experiences both hydrophilic interaction and ion-exchange interaction on the Click XIon material. According to the chromatographic evaluation, the optimized conditions were applied to HILIC-SPE to enrich GA from licorice, which leads to an increased content of GA from 13.67% to 64.22%. Finally, prep-RPLC was performed to obtain GA with purity higher than 99.00%，which demonstrating great prospect in large-scale preparation of GA.
authors.affiliation: School of Pharmaceutical Science & Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming, 650500, PR China.
authors.forename: Wenyi
authors.initials: W
authors.lastname: Yu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, PR China.
authors.forename: Hongli
authors.initials: H
authors.lastname: Jin
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, PR China.
authors.forename: Aijin
authors.initials: A
authors.lastname: Shen
authors.affiliation: School of Pharmaceutical Science & Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming, 650500, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Deng
authors.affiliation: School of Pharmaceutical Science & Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming, 650500, PR China.
authors.forename: Jianlian
authors.initials: J
authors.lastname: Shi
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, PR China.
authors.forename: Xingya
authors.initials: X
authors.lastname: Xue
authors.affiliation: School of Pharmaceutical Science & Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming, 650500, PR China. Electronic address: yadongkmmc@163.com.
authors.forename: Yadong
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, PR China. Electronic address: liuyanfang@dicp.ac.cn.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, PR China.
authors.forename: Xinmiao
authors.initials: X
authors.lastname: Liang
mesh_terms: D000893:Anti-Inflammatory Agents / Q000032:analysis / Q000302:isolation & purification*; D000998:Antiviral Agents / Q000032:analysis / Q000302:isolation & purification*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D006035:Glycyrrhiza / Q000737:chemistry*; D019695:Glycyrrhizic Acid / Q000032:analysis / Q000302:isolation & purification*; D057927:Hydrophobic and Hydrophilic Interactions; D052616:Solid Phase Extraction / Q000379:methods*
chemical_list: D000893:Anti-Inflammatory Agents; D000998:Antiviral Agents; D019695:Glycyrrhizic Acid
keywords: Glycyrrhizin; Hydrophilic interaction solid phase extraction; Licorice; Preparative reversed-phase liquid chromatography