甘氨胆酸中有关物质检查方法的研究

目的：建立HPLC-ELSD法测定甘氨胆酸的有关物质。 方法：采用Gemini C₁₈色谱柱（4.6 mm×150 mm，5 μm），以乙腈-0.1%甲酸（48：52）为流动相，流速0.5 mL·min^-1，ELSD检测，漂移管温度95℃，气体流速2.5 L·min^-1，Gain为8。 结果：在选定的条件下，甘氨胆酸与其中的5个杂质胆酸、甘氨胆酸乙酯、甘氨脱氧胆酸、甘氨脱氧胆酸乙酯、胆酸乙酯分离良好，它们的检测限相当的杂质浓度分别为0.008%、0.016%、0.016%、0.016%、0.022%、0.016%；5个杂质的平均回收率分别为100.5%、98.1%、105.7%、100.7%、101.4%；且上述6个物质浓度在3.75~31.25 μg·mL^-1范围内峰面积的对数与浓度的对数呈良好线性关系。 结论：经方法学验证，本法简便、专属、准确，可用于甘氨胆酸中有关物质的检查。

Objective: To establish an HPLC-ELSD method to determine the related substances in glycocholic acid. Methods: The HPLC method was performed on a Gemini C₁₈ column(4.6 mm×150 mm,5 μm) with the mobile phase of acetonitrile-0.1 %formic acid(48: 52).The flow rate was 0.5 mL·min^-1and the detector was Alltech ELSD 2000 (temperature of drift tube:95℃;airflow velocity;2.5 L·min^-1;Gain:8). Results: Under this specific condition,glycocholic acid and its five impurities(cholic acid,ethyl glycocholate,glycodeoxycholic acid,ethyl glycodeoxycholate,ethyl cholate) were well-separated.The limit of detection of glycocholic acid,cholic acid,ethyl glycocholate,glycodeoxycholic acid,ethyl glycodeoxycholate,ethyl cholate was 0.008%,0.016%,0.016%,0.016%,0.022%,0.016% respectively.The average recoveries of the five impurities were 100.5%,98.1%,105.7%,100.7%,101.4%.Linearity was observed within the range of 3.75-31.25 μg·mL^-1(r>0.9990) for all the analytes. Conclusion: The method was simple,specific,accurate and suitable for analyzing the related substances in glycocholic acid.

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