HPLC-MS/MS法测定老年糖尿病患者血浆中苯甲酸阿格列汀浓度

Determination of Alogliptin Benzoate Concentration in Plasma of Elderly Diabetic Patients by HPLC-MS/MS

目的:采用HPLC-MS/MS法测定老年糖尿病患者血浆中苯甲酸阿格列汀的浓度。方法:采用蛋白沉淀的方法处理样品,色谱柱为Diamonsil C18柱(4.6 mm×150 mm,5μm),使用乙腈-0.1%甲酸溶液作为流动相,进行梯度洗脱,流速0.6 mL·min^(-1),柱温30℃,进样量10μL。采用电喷雾离子源,正离子模式对血浆样品中的苯甲酸阿格列汀浓度进行定量检测,阿格列汀和西格列汀的检测离子对分别为m/z 340.0→116.0和m/z 408.0→235.0。结果:人血浆中苯甲酸阿格列汀在1~200 ng·mL^(-1)浓度范围线性关系良好,定量下限为1 ng·mL^(-1),批内、批间准确度分别为99.1%~108.5%、102.4%~109.2%,批内和批间的精密度RSD均<8.9%,平均提取回收率为86.4%~95.6%,基质效应为95.1%~106.7%。结论:本试验建立的方法简单,灵敏度和准确性高,适用于苯甲酸阿格列汀在老年糖尿病患者血浆中的含量测定。

Objective: To determine the plasma concentration of alogliptin benzoate in elderly patients with diabetes by HPLC-MS/MS. Methods: Protein precipitation was used to process the samples. The chromatographic column was a Diamonsil C18 column(4.6 mm×150 mm, 5 μm), and acetonitrile-0.1% formic acid water was used as the mobile phase. The flow rate was 0.6 mL·min^(-1), the column temperature was 30 °C, and the injection volume was 10 μL. The concentration of alogliptin benzoate in plasma samples was quantitatively detected by electrospray ion source and positive ion mode. The detection ion pairs of alogliptin and sitagliptin were respectively m/z 340.0→116.0, m/z 408.0→235.0. Results: The algagliptin in human plasma had a good linear relationship in the concentration range of 1-200 ng·mL^(-1). The lower limit of quantification was 1 ng·mL^(-1). The intra-batch and inter-batch accuracy were respectively 99.1%-108.5% and 102.4%-109.2%, RSD of the intra-batch and inter-assay precision was less than 8.9%, the average extraction recovery was 86.4%-95.6%, and the matrix effect was in the range of 95.1%-106.7%. Conclusion: The method established in this test is simple, sensitive,and highly accurate, and is suitable for the determination of alogliptin benzoate in elderly patients with diabetes plasma.