title: At-line hyphenation of high-speed countercurrent chromatography with Sephadex LH-20 column chromatography for bioassay-guided separation of antioxidants from vine tea (Ampelopsis grossedentata).
abstract: Vine tea (Ampelopsis grossedentata), a widely used healthy tea, beverage and herbal medicine, exhibited strong antioxidant activity. However, systematic purification of antioxidants, especially for those with similar structures or polarities, is a challenging work. Here, we present a novel at-line hyphenation of high-speed countercurrent chromatography with Sephadex LH-20 column chromatography (HSCCC-Sephadex LH-20 CC) for rapid and efficient separation of antioxidants from vine tea target-guided by 1,1-diphenyl-2-picryl-hydrazyl radical-high performance liquid chromatography (DPPH-HPLC) experiment. A makeup pump, a six-port switching valve and a trapping column were served as interface. The configuration had no operational time and mobile phase limitations between two dimensional chromatography and showed great flexibility without tedious sample-handling procedure. Seven targeted antioxidants were firstly separated by stepwise HSCCC using petroleum ether-ethyl acetate-methanol-water (4:9:4:9, v/v/v/v) and (4:9:5:8, v/v/v/v) as solvent systems, and then co-eluted antioxidants were on-line trapped, concentrated and desorbed to Sephadex LH-20 column for further off-line purification by methanol. It is noted that six elucidated antioxidants with purity over 95% exhibited stronger activity than ascorbic acid (VC). More importantly, this at-line hyphenated strategy could sever as a rapid and efficient pathway for systematic purification of bioactive components from complex matrix.
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, PR China.
authors.forename: Ruyi
authors.initials: R
authors.lastname: Ma
authors.affiliation: Food Science and Technology Program, Beijing Normal University-Hongkong Baptist University United International College, Zhuhai 519085, PR China.
authors.forename: Rongrong
authors.initials: R
authors.lastname: Zhou
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, PR China.
authors.forename: Runna
authors.initials: R
authors.lastname: Tong
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, PR China. Electronic address: shuyshi@gmail.com.
authors.forename: Shuyun
authors.initials: S
authors.lastname: Shi
authors.affiliation: College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, PR China.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Chen
mesh_terms: D032421:Ampelopsis / Q000737:chemistry*; D000975:Antioxidants / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D001713:Biphenyl Compounds / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D003377:Countercurrent Distribution / Q000295:instrumentation* / Q000379:methods; D003911:Dextrans / Q000737:chemistry*; D004867:Equipment Design; D005419:Flavonoids / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D010851:Picrates / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D000975:Antioxidants; D001713:Biphenyl Compounds; D003911:Dextrans; D005419:Flavonoids; D010851:Picrates; D010936:Plant Extracts; C025614:sephadex; C004931:1,1-diphenyl-2-picrylhydrazyl
keywords: Antioxidant; At-line hyphenation; HSCCC; Sephadex LH-20 CC; Vine tea