Purification of bufadieeolides from the skin of Bufo hufo gargarizans Cantor with positively charged C18 column

摘要

As a kind of promising anticancer compounds, the preparation of bufadienolides is a hot study spot:. Mow-ever, due to the complexity of biological sample, the purification of bufadienoiides 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 bufadienoiides from toad skin. By this method, the different ionic feature of the amino acid conjugated bufadienoiides (AACBs) and the free form bufadienoiides (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 bufadienoiides 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 bufadienoiides from toad skin.