RP-HPLC法测定蛇胆、鸡胆、猪胆的胆汁酸成分及其抑菌作用的研究

Study on Bile Acids from Bile Juice of Snake,Chicken and Pig by Reversed Phase Performance Liquid Chromatography and their Antibacterial Effects

建立了反相液相色谱法同时测定动物胆汁样品中的牛磺胆酸、牛磺去氧胆酸、牛磺鹅去氧胆酸、甘氨石胆酸等结合型胆汁酸成分的方法。采用色谱柱为Agilent SB-C18柱(4.6×250 mm,5μm),流动相为乙腈-25mM磷酸二氢钾盐溶液(30:70),检测波长200nm。流速1.0mL/min,柱温40℃。各组分在一定范围内有良好的线性关系。本法具有简单快速、分离效果好、重复性好、回收率高的优点。同时研究了动物胆汁及其主要胆汁酸成分的体外抑菌作用,结果表明胆汁酸对于金黄色葡萄球菌的生长有明显的抑制作用,且牛磺结合型胆汁酸TCA的抑菌作用强于游离型胆汁酸CA。

A reversed phase high performance liquid chromatographic method for the determination of conjugated bile acids in bile juice was developed. An Agilent SB - C18 column（4.6 × 250 mm,5μm）was selected as separation. TCA, TCDCA, TDCA, GLCA could be separated in 30minites and then detected at 200nm, using acetonitrile potassium dihydrogen phosphate buffer as the mobile phase at flow rate 1.0 mL/min. Linear calibration graphs for each component were obtained within their range. This method proved to be simple, rapid with good separations and good recoveries. The antibacterial effects of bile acids were also studied in this research and the results showed that both TCA and CA can apparently inhibit the Staphylococcus Aureus in vitro; furthermore, TCA＇s antibacterial effects are higher than CA＇s under the same concentrations.