UFLC-MS/MS法测定胎盘灌流液中格列本脲浓度

UFLC-MS/MS determination of glibenclamide in placental perfusate

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.