Purification of bufadienolides from the skin of Bufo bufo gargarizans Cantor with positively charged C18 column

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• 作者：Xiaolong Li^a ; ^b ; ^c ; Zhimou Guo^a ; Chaoran Wang^a ; Aijin Shen^a ; Yanfang Liu^a ; ^{liuyanfang@dicp.ac.cn" class="auth_mail} ; Xiuli Zhang^a ; Weijie Zhao^b ; Xinmiao Liang^a ; ^{liangxm@dicp.ac.cn" class="auth_mail}
• 关键词：Purification ; Bufadienolides ; Bufo bufo gargarizans Cantor ; Reversed-phase liquid chromatography ; Positively charged C18
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：15 April, 2014
• 年：2014
• 卷：92
• 期：Complete
• 页码：105-113
• 全文大小：895 K

文摘

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.