title: Purification of bufadienolides from the skin of Bufo bufo gargarizans Cantor with positively charged C18 column.
abstract: As a kind of promising anticancer compounds, the preparation of bufadienolides is a hot study spot. However, due to the complexity of biological sample, the purification of bufadienolides from a crude sample (toad skin) is a tough work. In this paper, we reported a new way based on positively charged C18 material (XCharge C18) to quickly separate and purify bufadienolides from toad skin. By this method, the different ionic feature of the amino acid conjugated bufadienolides (AACBs) and the free form bufadienolides (AAUBs) was firstly utilized to obtain distinct separation selectivity on the XCharge C18 column. Additionally, the peak tailing problem of AACBs on conventional C18 was resolved and better resolutions were achieved on the XCharge C18, thus, two kinds of bufadienolides on one column were successfully purified respectively. Taking F13 as an example, the method was validated by liquid chromatography-mass spectrometry (LC-MS), and then 4 AACBs as well as 4 AAUBs were simultaneously purified by preparative XCharge C18. In addition, the application of this method in other fractions was also validated. The results suggested that the developed method is a practical and promising tool for efficient separation and purification of bufadienolides from toad skin.
authors: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; School of Pharmaceutical Science and Technology, State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors: Xiaolong
authors: X
authors: Li
authors: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors: Zhimou
authors: Z
authors: Guo
authors: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors: Chaoran
authors: C
authors: Wang
authors: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors: Aijin
authors: A
authors: Shen
authors: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. Electronic address: liuyanfang@dicp.ac.cn.
authors: Yanfang
authors: Y
authors: Liu
authors: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors: Xiuli
authors: X
authors: Zhang
authors: School of Pharmaceutical Science and Technology, State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China.
authors: Weijie
authors: W
authors: Zhao
authors: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. Electronic address: liangxm@dicp.ac.cn.
authors: Xinmiao
authors: X
authors: Liang
mesh_terms: D000818:Animals; D002018:Bufanolides / Q000737:chemistry*; D002023:Bufo bufo / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D011724:Pyridazines / Q000737:chemistry*; D012867:Skin / Q000737:chemistry*
chemical_list: D002018:Bufanolides; D011724:Pyridazines; C006149:minaprine
keywords: Bufadienolides; Bufo bufo gargarizans Cantor; Positively charged C18; Purification; Reversed-phase liquid chromatography