title: Efficient methods for isolating five phytochemicals from Gentiana macrophylla using high-performance countercurrent chromatography.
abstract: Efficient high-performance countercurrent chromatography methods were developed to isolate five typical compounds from the extracts of Gentiana macrophylla. n-Butanol-soluble extract of G. macrophylla contained three hydrophilic iridoids, loganic acid (1), swertiamarin (2) and gentiopicroside (3), and a chromene derivative, macrophylloside D (4) which were successfully isolated by flow rate gradient (1.5 mL/min in 0-60 min, 5.0 mL/min in 60-120 min), and consecutive flow rate gradient HPCCC using n-butanol/0.1% aqueous trifluoroacetic acid (1:1, v/v, normal phase mode) system. The yields of 1-4 were 22, 16, 122, and 6 mg, respectively, with purities over 97% in a flow rate gradient high-performance countercurrent chromatography, and consecutive flow rate gradient high-performance countercurrent chromatography gave 1, 2, 3 (54, 41, 348 mg, respectively, purities over 97%) and 4 (13 mg, purity at 95%) from 750 mg of sample. The main compound in methylene chloride soluble extract, 2-methoxyanofinic acid, was successfully separated by n-hexane/ethyl acetate/methanol/water (4:6:4:6, v/v/v/v, flow-rate: 4 mL/min, reversed phase mode) condition. The structures of five isolates were elucidated by1H,13C NMR and ESI-Q-TOF-MS spectroscopic data which were compared with previously reported values.
authors: College of Pharmacy and Integrated Research Institute of Pharmaceutical Sciences, The Catholic University of Korea, Bucheon, Republic of Korea.
authors: Taewoong
authors: T
authors: Rho
authors: College of Pharmacy and Integrated Research Institute of Pharmaceutical Sciences, The Catholic University of Korea, Bucheon, Republic of Korea.
authors: Mila
authors: M
authors: Jung
authors: Laboratory of Pharmacognosy and Natural Product Derived Medicine, College of Pharmacy, Chung-Ang University, Seoul, Republic of Korea.
authors: Min Won
authors: MW
authors: Lee
authors: College of Pharmacy, Dongguk University-Seoul, Goyang, Republic of Korea.
authors: Young-Won
authors: YW
authors: Chin
authors: College of Pharmacy and Integrated Research Institute of Pharmaceutical Sciences, The Catholic University of Korea, Bucheon, Republic of Korea.
authors: Kee Dong
authors: KD
authors: Yoon
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution; D030012:Gentiana / Q000737:chemistry*; D006586:Hexanes; D064209:Phytochemicals / Q000302:isolation & purification*; D010936:Plant Extracts / Q000737:chemistry*
chemical_list: D006586:Hexanes; D064209:Phytochemicals; D010936:Plant Extracts; C026385:n-hexane
keywords: Chromens; Gentiana macrophylla; High-performance countercurrent chromatography; Iridoids