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论文类型：期刊论文

发表时间：2014-08-25

发表刊物：JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS

收录刊物：SCIE、PubMed、Scopus

卷号：97

页面范围：54-64

ISSN号：0731-7085

关键字：Active bufadienolides; Hydrophilic solid-phase extraction; Class separation; Purification; Toad skin

摘要：Traditional Chinese medicines (TCMs) have played a significant role in the process of discovering natural bioactive compounds, especially in anticancer therapeutics. However, the components of TCMs are complex mixtures with wide variation in polarity and content, which leads to inefficiency in the process of active compound discovery from TCMs. In this paper, the popular strategy of utilizing "pre-fractionated natural product libraries" has been improved by a new class separation approach to accelerate the process. As an example, the skin of Bufo bufo gargarizans Cantor, a well-known TCM, mainly contains two distinct bufadienolide classes: amino acid-conjugated bufadienolides (AACBs) and free form bufadienolides (AAUBs). We utilized hydrophilic interaction liquid chromatography solid-phase extraction (HILIC-SPE) to resolve the two types of bufadienolides, which co-eluted on C18 columns. By this strategy, twelve bufadienolides of the two types were purified via prep-HPLC from one active fraction, and eight of them were identified by H-1 NMR and C-13 NMR. These results indicated that the class separation method not only overcame the limited orthogonality in a 2D-RPLC x RPLC system but also accelerated the process of active compound discovery. (c) 2014 Elsevier B.V. All rights reserved.