HPLC-ELSD分析米格列醇和杂质1-脱氧野尻霉素

HPLC-ELSD method for analyzing miglitol and its impurities of 1-deoxynojirimycin

目的:建立一种高效液相-蒸发光散色检测法用于米格列醇和1-脱氧野尻霉素的杂质分析。方法:使用随行标准曲线法测定杂质含量,采用Inertsil NH2(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-15 mmol.L-1pH 5.5乙酸铵(75∶25),流速为1.2 mL.min-1,柱温为35℃;Alltech ELSD2000检测器的漂移管温度为90℃,载气流速为2.9 L.min-1。结果:米格列醇和1-脱氧野尻霉素在0.02～5.05 mg.mL-1范围内峰面积对质量浓度呈现良好的对数线性关系,r分别为0.9985和0.9989;定量限和检出限分别为1和0.4μg;两种化合物在该色谱条件下重现性良好;精密度n=6时,RSD分别为1.5%和1.0%。结论:本方法有效、快速、简便、结果可靠,可用于米格列醇和1-脱氧野尻霉素的杂质分析。

Objective： To develop a HPLC-ELSD method for impurity test of miglitol and 1-deoxynojirimycin（1-DNJ）.Methods： The impurities were determined by a standard curve method.Inertsil Amide column（250mm×4.6mm,5μm） was used as the HPLC column.The separation was performed using a mixture of acetonitrile and distilles water（75∶25,containing 15mmol·L-1 ammonium acetate,pH 5.5）.The flow rate was adjusted at 1.2mL·min-1,column temperature is 35℃.One of split eluents was sent to an Alltech ELSD 2000 evaporative light scattering detector.The temperature of the drift tube was set at 90℃,and the flow of gas at 2.9L·min-1.Results： A good log-linear correlation was found between the peak areas and concentrations of miglitol and 1-deoxynojirimycin（0.02~5.05mg·mL-1）.The correlation coefficients were 0.9985 and 0.9989 for miglitol and 1-deoxynojirimycin,respectively.The quantitative limit and detectable limit were 1 and 0.4μg,respectively.The two compounds had good reproducibility in the chromatographic conditions.When the precision was equal to 6,RSDs were 1.5% and 1.0%.Conclusion： The improved HPLC-ELSD method is effective,fast and simple.The results are reliable and accurate,so it can be used for impurity test of miglitol and 1-deoxynojirimycin.