HPLC-CAD法测定6-乙基鹅去氧胆酸起始物料中有关物质

Determination of Related Substances in the Starting Material of 6-Ethylchenodeoxycholic Acid by HPLC-CAD

目的:使用电喷雾检测器建立6-乙基鹅去氧胆酸起始物料的有关物质检测方法。方法:采用Waters XBridge Shield RP18(150 mm×4.6 mm,3.5μm)色谱柱,以0.05%三氟乙酸水溶液为流动相A,以乙腈为流动相B,梯度洗脱,流速为1.4 mL·min-1,柱温为35℃,电喷雾检测器(雾化温度:35℃)。结果:本品主峰与相邻杂质分离良好,在质量浓度0.5~15μg·mL^(-1)范围内线性关系良好,r=0.999,定量限为13.2 ng,检测限为5.3 ng,杂质重复性考察结果RSD均小于2%(n=6)。结论:方法精密度好、准确度高,可用于本品的有关物质检测。

Objective:To establish an method for the determination of related substances in the starting material of 6-ethylchenodeoxycholic acid by Charged Aerosol Detection(CAD).Methods:Waters XBridge Shield RP18 column(150 mm×4.6 mm,3.5μm)was used,0.05%Trifluoroacetic acid in water was used as mobile phase A,with Acetonitrile as mobile phase B in a gradient elution.The flow rate was 1.4 mL·min-1,column temperature was 35℃,CAD was used with atomization temperature 35℃.Results:The main substance and its impurities could be well separated.A good linear was obtained between ranges form 0.5μg·mL^(-1) to 15μg·mL^(-1) with Correlation coefficient 0.999.The limit of quantification was 13.2 ng,the limit of detection was 5.3 ng,and the RSD(n=6)of the impurity repeatability was less than 2%.Conclusion:The method has good precision and high accuracy,and can be used for the detection of related substances in the starting material of 6-ethylchenodeoxycholic acid.