目的: 建立采用高效液相色谱法测定依折麦布有关物质。方法: 采用Agilent TC-C18(250 mm×4.6 mm,5.0 μm)色谱柱,以0.05 mol·L-1磷酸二氢钾溶液-乙腈-四氢呋喃(60∶20∶20)为流动相,流速1.0 mL·min-1,柱温35 ℃,检测波长232 nm,进样体积20 μL。结果: 依折麦布峰与各杂质峰分离良好;依折麦布峰与杂质A、杂质B、杂质C、杂质D、杂质E与杂质F的线性范围分别为0.409~4.090 μg·mL-1(r=0.995 0)、0.402~4.020 μg·mL-1(r=0.998 1)、0.408~4.080 μg·mL-1(r=0.999 1)、0.411~4.110 μg·mL-1(r=0.999 3)、0.4050~4.050 μg·mL-1(r=0.998 1)、0.541~5.41 μg·mL-1(r=0.998 5)、0.657~6.57 μg·mL-1(r=0.997 5);检测限分别为0.130、0.135、0.135、0.140、0.140、0.180、0.210 μg·mL-1,定量限分别为0.409、0.402、0.408、0.411、0.405、0.541、0.657 μg·mL-1;杂质A、杂质B、杂质C、杂质D、杂质E与杂质F的平均回收率分别为99.4%、101.2%、100.1%、101.7%、100.0%、101.0%,RSD分别为0.85%、1.1%、1.0%、0.62%、0.87%、0.72%。3批依折麦布样品检测结果显示,已知杂质及其他最大单个杂质含量均＜0.10%,杂质总含量＜0.50%。结论: 建立的高效液相色谱法方法可靠、准确,可用于依折麦布有关物质的测定。
Objective: To develop a method for related substances determination of ezetimibe by HPLC. Methods: Agilent TC-C18 column (250 mm×4.6 mm, 5.0 μm) was used. The mobile phase consisted of 0.05 mol·L-1 potassium dihydrogen phosphate solution-acetonitrile-tetrahydrofuran(60∶20∶20) with a flow rate of 1.0 mL·min-1. The column temperature was 35 ℃, the detection wavelength was 232 nm,and the injection volume was 20 μL. Results: Chromatographic peaks of ezetimibe and impurities were separated. Linear ranges of ezetimibe, and impurities A, B, C, D, E and F were 0.409-4.090 μg·mL-1(r=0.995 0), 0.402-4.020 μg·mL-1(r=0.998 1), 0.408-4.080 μg·mL-1(r=0.999 1), 0.411-4.110 μg·mL-1(r=0.999 3), 0.405 0-4.050 μg·mL-1(r=0.998 1), 0.541-5.41 μg·mL-1(r=0.998 5), 0.657-6.57 μg·mL-1 (r=0.997 5), respectively; LODs were 0.130, 0.135, 0.135, 0.140, 0.140, 0.180, 0.210 μg·mL-1; LOQs were 0.409, 0.402, 0.408, 0.411, 0.405, 0.541, 0.657 μg·mL-1. Average recoveries of impurities A, B, C, D, E and F were 99.4%, 101.2%, 100.1%, 101.7%, 100.0%, 101.0%, and RSDs were 0.85%, 1.1%, 1.0%, 0.62%, 0.87%, 0.72%. The detection results of 3 batches of ezetimibe samples showed that the contents of known impurities and other maximum single impurity were less than 0.10%, and the total content of impurities was all less than 0.50%. Conclusion: The method is accurate and reliable for determination of related substances of ezetimibe.
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