HPLC-ELSD法测定猪胆粉中的甘氨猪去氧胆酸和甘氨鹅去氧胆酸的含量

摘要

目的: 建立HPLC-ELSD法测定猪胆粉中甘氨猪去氧胆酸和甘氨鹅去氧胆酸的含量测定方法.方法: 猪胆粉甲醇提取液采用Welch Ultimate Plus C 18色谱柱(250 mm× 4.6 mm,5 μm),以乙腈-0.1%甲酸溶液(40∶60)为流动相,流速为1.0 ml·min-1,检测器为奥泰ELSD 2000ES,漂移管温度为105℃,氮气流速为每分钟2.5 L.结果: 甘氨猪去氧胆酸和甘氨鹅去氧胆酸分别在 0.306 ~ 4.590μg (r=0.999 5)、 0.280 ~ 4.194μg(r=0.999 4)范围内呈现良好的线性关系,其平均回收率分别为99.23%, 100.37%, RSD 分别为2.0%, 1.8% (n=9).结论: 该方法结果准确,灵敏度高,重复性好,可用于猪胆粉的质量 控制.%Objective: To establish a method for the simultaneous determination of glycohyodeoxycholic acid and glycochenodeoxycholic acid in pulvis fellis suis by HPLC-ELSD.Methods: The analysis of methanol extract of pulvis fellis suis was performed on a Welch Ultimate Plus C 18 (250 mm× 4.6 mm ,5 μm)column, and the mobile phase consisted of acetonitrile and 0.1% formic acid (40∶60).The flow rate was 1.0 ml ·min-1.Alltech ELSD 2000ES was used as the detector, the drift tube temperature was 105℃ and the nitrogen flow rate was 2.5 L·min-1.Results: Glycohyodeoxycholic acid and glycochenodeoxycholic acid had good linear relationships within the range of 0.306-4.590 μg (r=0.999 5) and 0.280-4.194 μg (r=0.999 4), respectively.The average recovery was 99.23% with RSD of 2.0% and 100.37% with RSD of 1.8% (n =9), respectively.Conclusion: The method is accurate, sensitive and repeatable, which can be used for the quality control of pulvis fellis suis.