Separation and purification of bovine serum albumin binders from Fructus polygoni orientalis using off-line two-dimensional complexation high-speed counter-current chromatography target-guided by ligand fishing

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• 作者：Qi Liu ; Shuyun Shi ; Liangliang Liu ; Hua Yang ; Wen Su ; Xiaoqing Chen
• 关键词：Fructus polygoni orientalis ; Off-line two-dimensional high-speed counter-current chromatography ; HPLC&ndash ; MS/MS ; Target-guided ; HSCCC
• 刊名：Journal of Chromatography A
• 出版年：2013
• 出版时间：23 August, 2013
• 年：2013
• 卷：1304
• 期：Complete
• 页码：183-193
• 全文大小：2023 K

文摘

In this study, off-line two-dimensional (2D) complexation high speed counter-current chromatography (HSCCC) was developed for the separation of bovine serum albumin (BSA) binders from the ethyl acetate extract of Fructus polygoni orientalis. Target-guided strategy of BSA functionalized iron oxide magnetic nanoparticles coupled with high performance liquid chromatography-tandem mass spectrometry ((BSA-Fe₃O₄ MNPs)-HPLC-MS/MS) experiment was proposed. In the orthogonal separation system, a Normal-Phase HSCCC with 0.01 mol/L copper ion as complexation agent in the aqueous phase was employed for the first dimension and Recycling HSCCC, Reverse-Phase HSCCC with 0.1 mol/L copper ion were used for the second dimension in parallel. Including two pairs of cis-trans isomers, seven BSA binders including 3,5,7-Trihydroxychromone (1), taxifolin (2), N-cis-paprazine (3), N-cis-feruloyltyramine (4), N-trans-paprazine (5), N-trans-feruloyltyramine (6) and an unidentified compound (7) were obtained. The purities of these seven compounds were all over 95.0 % as determined by HPLC. The complexation HSCCC behaviors of seven compounds were also investigated by studying their relationship with copper ion. Results showed that the combinative method using (BSA-Fe₃O₄ MNPs)-HPLC and HSCCC is a quick, efficient, and reproductive technique to isolate potentially bioactive compounds from the complex mixture system of natural products. And the usage of off-line 2D-HSCCC and introduction of chelating metal ion into solvent system are effective ways to implement HSCCC separations in complex samples.