目的：建立并验证一种定性定量检测人血浆中5种常见磺酰脲降糖药的方法，并将其应用于临床检测。方法：血浆样品经乙腈沉淀蛋白后，采用液相-串联质谱联用仪测定。色谱柱为BEH C18，以含0.01%甲酸乙腈溶液-含0.01%甲酸水溶液为流动相梯度洗脱，流速为0.4 mL·min^-1，电喷雾正离子模式，多反应监测。结果：格列齐特、格列吡嗪、格列美脲、格列本脲、格列喹酮质量浓度分别在0.037~9（r=0.998 3）、0.037~9（r=0.999 2），0.037~9（r=0.992 8），0.037~9（r=0.996 8），0.037~9（r=0.998 8）μg·mL^-1内线性关系良好；定量下限分别为0.004 6，0.009 2，0.004 6，0.004 6，0.009 2 μg·mL^-1；日内和日间精密度均小于<13.88%，相对回收率均在77.57%~100.83%之间；提取回收率均在69.69%~91.54%之间；基质效应均在84.32%~102.86%之间；在多种储存条件下，处理前后的血浆样品稳定性好。本方法应用于1例2岁小儿格列本脲中毒检测，结果表明当血浆中格列本脲浓度为6.2，0.21，0.09 μg·mL^-1时，均呈现降低血糖作用。结论：该方法检测速度快、专属性高、重复性好，可用于磺酰脲类药物中毒检测。经临床实践证明，本方法提供的检测数据可作为磺酰脲中毒的重要临床指标，对处于不同治疗阶段的中毒患者，均具有重要的临床指导意义。

OBJECTIVE To establish and validate a method for simultaneous detection of 5 common sulfonylureas in human plasma, which can be applied to clinical examination. METHODS The plasma samples were measured by liquid phase-tandem mass spectrometry after their proteins were precipitated by acetonitrile. The chromatographic column was BEH C18. Water solution with 0.01% formic acid and 0.01% acetonitrile solution were used as mobile phase to perform gradient elution. The flow rate was 0.4 ml·min^-1. MRM and ESI positive ion mode were used for monitoring. RESULTS The linear ranges of gliclazide, glipizide, glimepiride, glyburide, gliquidone were 0.037-9 (r=0.998 3),0.037-9(r=0.999 2),0.037-9(r=0.992 8),0.037-9(r=0.996 8),0.037-9(r=0.998 8)μg·mL^-1, respectively. The minimum quantitative limits were 0.004 6, 0.009 2, 0.004 6, 0.004 6, 0.009 2 μg·mL^-1, respectively. The RSDs of inter-day and intra-day precision were both less than <13.88%. The relative recovery were 77.57%-100.83%. The extraction recovery rates were 69.69%-91.54%. The matrix effects were 84.32%-102.86%. Plasma samples were stable before and after treatment under various storage conditions. This method has been successfully applied in one case of 2 years old children intoxicated by glyburide. The results showed that glibenclamide showed hypoglycemic effects, at the plamsa concentrations of 6.2, 0.21, 0.09 μg·mL^-1. CONCLUSION This method is rapid, specific and reproducible, which can be used for clinical detection of sulfonylureas. It was proved that the testing data provided by this method could be used as an important clinical diagnosis indicator of sulfonylurea poisoning, which has important clinical guiding significance for poinsoned patients at different treatment stages.