目的:建立测定胎盘灌流液中格列本脲浓度的超快速液相色谱-串联质谱(UFLC-MS/MS)方法。方法:以格列齐特为内标,胎盘灌流液样品用乙腈沉淀蛋白后进样分析。采用Shim-pack XR-ODS II C18(2.0 mm×75 mm,2.2 μm)色谱柱,以0.1%(v/v)甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱(0~0.5 min,90%A,10%B;0.5~1.5 min,90%A→10%A,10%B→90%B;1.5~2.7 min,10%A,90%B;2.7~3.5 min,90%A,10%B),流速0.4 mL·min^-1。质谱采用多反应监测(MRM)扫描模式,以电喷雾离子源(ESI)在正离子电离模式下进行测定,格列本脲和格列齐特的定量分析离子分别为m/z 494.0→369.0和m/z 324.1→110.1。结果:胎盘灌流液中格列本脲浓度在0.25~250 ng·mL^-1范围内线性关系良好(r=0.9998),日内及日间RSD均小于15%,稳定性考察结果良好。结论:该方法快速、灵敏,专属性强,可用于胎盘灌流液中格列本脲浓度的测定。
Objective: To establish a UFLC-MS/MS method for the determination of glibenclamide in placental perfusate. Methods: Samples were precipitated with acetonitrile.Analysis of glibenclamide and gliclazide(internal standard)were carried out on a Shim-pack XR-ODS II C18 column(2.0 mm×75 mm,2.2 μm).Gradient elution(0-0.5 min,90%A,10%B;0.5-1.5 min,90%A→10%A,10%B→90%B;1.5-2.7 min, 10%A,90%B;2.7-3.5 min,90%A,10%B) was adopted using water(containing 0.1% formic acid,v/v)(A) and acetonitrile(B) at a flow rate of 0.4 mL·min^-1.Detection was performed with multiple reaction monitoring(MRM) using positive electrospray ionization(ESI) at m/z 494.0→369.0 for glibenclamide and m/z 324.1→110.1 for gliclazide. Results: The calibration curve of glibenclamide was linear over the range of 0.25-250 ng·mL^-1 (r=0.9998).Inter- and intra-day RSDs were less than 15%,and the analytes were proved to be stable. Conclusion: This method is rapid,sensitive and specific,and can be applied to the determination of glibenclamide in placental perfusate.