title: On-line comprehensive two-dimensional normal-phase liquid chromatography×reversed-phase liquid chromatography for preparative isolation of toad venom.
abstract: An on-line comprehensive preparative two-dimensional normal-phase liquid chromatography×reversed-phase liquid chromatography (2D NPLC×RPLC) system was constructed with a newly developed vacuum evaporation assisted adsorption (VEAA) interface, allowing fast removal of NPLC solvent in the vacuum condition and successfully solving the solvent incompatibility problem between NPLC and RPLC. The system achieved on-line solvent exchange within the two dimensions and its performance was illustrated by gram-scale isolation of crude extract from the venom of Bufo bufo gargarizans. Within separation time of ∼20h, 19 compounds were obtained with high purity in a single run. With the VEAA interface, the 2D system exhibited apparent advantages in separation efficiency and automation compared with conventional methods, indicating its promising application in the routine separation process for complicated natural products.
authors.affiliation: School of Pharmaceutical Sciences, Xiamen University, South Xiang-An Road, Xiamen 361102, China.
authors.forename: Jia-Fu
authors.initials: JF
authors.lastname: Li
authors.affiliation: Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography State Oceanic Administration, Xiamen 361005, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Fang
authors.affiliation: School of Pharmaceutical Sciences, Xiamen University, South Xiang-An Road, Xiamen 361102, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Yan
authors.affiliation: Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807, Taiwan, China.
authors.forename: Fang-Rong
authors.initials: FR
authors.lastname: Chang
authors.affiliation: School of Pharmaceutical Sciences, Xiamen University, South Xiang-An Road, Xiamen 361102, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wu
authors.affiliation: School of Pharmaceutical Sciences, Xiamen University, South Xiang-An Road, Xiamen 361102, China. Electronic address: wuyl@xmu.edu.cn.
authors.forename: Yun-Long
authors.initials: YL
authors.lastname: Wu
authors.affiliation: School of Pharmaceutical Sciences, Xiamen University, South Xiang-An Road, Xiamen 361102, China. Electronic address: qyk@xmu.edu.cn.
authors.forename: Ying-Kun
authors.initials: YK
authors.lastname: Qiu
mesh_terms: D000327:Adsorption; D000664:Amphibian Venoms / Q000302:isolation & purification*; D000818:Animals; D001331:Automation; D002018:Bufanolides / Q000737:chemistry / Q000302:isolation & purification; D002023:Bufo bufo; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D009682:Magnetic Resonance Spectroscopy; D021241:Spectrometry, Mass, Electrospray Ionization; D013696:Temperature
chemical_list: D000664:Amphibian Venoms; D002018:Bufanolides
keywords: Bufadienolides; Bufo bufo gargarizans; Toad venom; Two dimensional normal-phase liquid chromatography×reversed-phase liquid chromatography; Vacuum evaporation assisted adsorption interface