HPLC法测定格列美脲中间体的有关物质

Analysis of related substances in glimepiride intermediate by HPLC

目的:建立HPLC法测定格列美脲原料药中间体的有关物质。方法:采用五氟苯基键合硅胶(150 mm×4.6 mm,2.7μm)色谱柱,以磷酸二氢铵缓冲溶液-乙腈为流动相,等度洗脱,流速0.5 mL·min^(-1),柱温25℃,检测波长225 nm。结果:格列美脲中间体与4个已知杂质A~D色谱峰均能良好分离,并分别在0.008~6.024μg·mL^(-1)(r=0.9999)、0.005~6.013μg·mL^(-1)(r=0.9996)、0.011~5.987μg·mL^(-1)(r=0.9998)、0.012~6.045μg·mL^(-1)(r=0.9998)、0.029~5.989μg·mL^(-1)(r=0.9996)范围内线性关系良好(n=6)。格列美脲中间体、杂质A、杂质B、杂质C、杂质D的定量限分别为8.3、5.0、10.6、11.9、29.4 ng·mL^(-1);杂质A~D平均回收率(n=9)分别为107.0%、100.0%、98.0%、101.6%。6批格列美脲中间体样品测定结果显示,已知杂质及其他最大单个杂质含量均小于0.50%,杂质总量小于1.0%。结论:经方法学验证,本方法灵敏、快速,专属性强,准确度高,可用于格列美脲原料药中间体有关物质的测定。

Objective:To establish an HPLC method for the determination of the related substances in glimepiride intermediate.Methods:The determination was performed on a pentafluorophenyl column(150 mm×4.6 mm,2.7μm).The mobile phase consisted of ammonium dihydrogen phosphate buffer solution and acetonitrilewith isocratic elution.The flow rate was 0.5 mL·min^(-1),and the column temperature was 25℃,the detection wavelengthwas set at 225 nm.Results:The resolutions between the peaks of glimepiride intermediate and four knownimpurities(including impurities A-D)were good.The calibration curves of glimepiride intermediate and the known impurities were linear in the ranges of 0.008-6.024μg·mL^(-1)(r=0.9999)、0.005-6.013μg·mL^(-1)(r=0.9996),0.011-5.987μg·mL^(-1)(r=0.9998),0.012-6.045μg·mL^(-1)(r=0.9998)、0.029-5.989μg·mL^(-1)(r=0.9996).The limits of quantitation were 8.3,5.0,10.6,11.9,29.4 ng·mL^(-1).The recoveries(n=9)were 103.2%,100.0%,98.6%,99.9%for the impurities A-D,respectively.The determination results of the sixbatches of glimepiride intermediate samples showed that the known impurities were less than 0.10%,the maximum single unknown impurity was less than 0.10%,and the total impurity was less than 0.15%.Conclusion:The method is proved to be simple,rapid and accurate,and can be used for thedetermination of relatedsubstances in Glimepiride intermediate.