Li Sainan, Liu Chunming, Guo Liping, Zhang Yuchi, Wang Jing, Ma Bing, Wang Yueqi, Wang Yumeng, Ren Junqi, Yang Xiaojing, Qin Yao, Tang Ying
Central Laboratory, Changchun Normal University, Changchun, China; Faculty of Chemistry, Northeast Normal University, Changchun, China.
J Sep Sci. 2014 Sep;37(18):2504-12. doi: 10.1002/jssc.201400475. Epub 2014 Aug 4.
Cortex Phellodendri is a typical Chinese herb with a large number of alkaloids existing in all parts of it. The most common methods for screening and isolating alkaloids are mostly labor intensive and time consuming. In this study, a new assay based upon ultrafiltration liquid chromatography was developed for the rapid screening of ligands for α-glucosidase and xanthine oxidase. The C. Phellodendri extract was found to contain two alkaloids with both α-glucosidase- and xanthine oxidase binding activities and one lactone with α-glucosidase-binding activity. Subsequently, with the help of high-speed countercurrent chromatography, the specific binding ligands including palmatine, berberine, and obaculactone with purities of 97.38, 96.12, and 96.08%, respectively, were successfully separated. An optimized low-toxicity two-phase solvent system composed of ethyl acetate/n-butanol/ethanol/water (3.5:1.7:0.5:5, v/v/v/v) was used to isolate the three compounds mentioned above from C. Phellodendri. The targeted compounds were identified by liquid chromatography coupled with mass spectrometry and NMR spectroscopy. Therefore, ultrafiltration liquid chromatography combined with high-speed countercurrent chromatography is not only a powerful tool for screening and isolating α-glucosidase and xanthine oxidase inhibitors in complex samples but is also a useful platform for discovering bioactive compounds for the prevention and treatment of diabetes mellitus and gout.
黄柏是一种典型的中药材，其各个部位均含有大量生物碱。筛选和分离生物碱的最常用方法大多 labor intensive 且耗时。在本研究中，开发了一种基于超滤液相色谱的新方法，用于快速筛选α-葡萄糖苷酶和黄嘌呤氧化酶的配体。发现黄柏提取物含有两种具有α-葡萄糖苷酶和黄嘌呤氧化酶结合活性的生物碱以及一种具有α-葡萄糖苷酶结合活性的内酯。随后，借助高速逆流色谱法，成功分离出纯度分别为97.38%、96.12%和96.08%的巴马汀、小檗碱和奥巴库内酯等特异性结合配体。使用由乙酸乙酯/正丁醇/乙醇/水（3.5:1.7:0.5:5，v/v/v/v）组成的优化低毒两相溶剂系统从黄柏中分离上述三种化合物。通过液相色谱-质谱联用和核磁共振光谱对目标化合物进行鉴定。因此，超滤液相色谱与高速逆流色谱相结合不仅是筛选和分离复杂样品中α-葡萄糖苷酶和黄嘌呤氧化酶抑制剂的有力工具，也是发现用于预防和治疗糖尿病和痛风的生物活性化合物的有用平台。
