高效液相色谱法测定胎盘灌流液中格列苯脲的浓度

Determination of Glibenclamide Concentration in Placental Perfusion Medium by High Performance Liquid Chromatography

目的建立测定胎盘灌流液中格列苯脲浓度的高效液相色谱(HPLC)检测方法。方法采用的色谱柱为Symmetry Shield RP C18(150 mm×4.6 mm,5μm),柱温40℃,流动相为NaH2PO4缓冲盐(25 mmol/L,pH值5.2)︰乙腈=1︰1;内标为格列齐特,流速1.0 mL/min,检测波长228 nm,采用液-液萃取预处理方法测定胎盘灌流液中格列苯脲的浓度。结果格列苯脲浓度线性范围为2.0～25.0μg/mL,线性方程为:y=0.226x+0.002,r=0.999 9(n=6),日内相对标准偏差(RSD)<3.1%,日间RSD<9.5%,方法学回收率为95.32%～103.35%。结论 HPLC检测方法灵敏、简便,可用于胎盘灌流液中格列苯脲浓度的检测。

Objective To establish a high performance liquid chromatography （HPLC） method for determining the concentration of glibenclamide in placental perfusion medium. Methods The liquid chromatography column was Symmetry Shield RP C18 （150 mm×4.6 mm, 5 μm） at 40℃ . The mobile phase consisted of a mixture of NaH2PO4 （25 mmol/L, pH 5.2）-acetonitrile （1 ： 1, V/V）. The flow rate was 1.0 mL/min. UV detection wavelength was set at 228 nm. A liquid-liquid extraction pretreatment method was used to detect the concentration of glibenclamide placental perfusion medium. Results A linearity was obtained from 2.0 to 25.0 μg/mL for glibenclamide. The calibration equation of glibenclamide was y=0.226x＋0.002, r=0.999 9 （n=6）. The relative standard deviations within day and between days were less than 3.1% and less than 9.5% for glibenclamide. The mean recoveries of glibenclamide were 95.32% to 103.35%. Conclusion This method can be used for determining glibenclamide concentration in placental perfusion medium.