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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 the determination of glibenclamide in placental perfusate. The column was Symmetry Shield RP C18 (150 mm × 4.6 mm, 5 μm). The column temperature was 40 ℃ and the mobile phase was NaH2PO4 buffer (25 mmol / L, pH 5.2) Gliclazide, the flow rate was 1.0 mL / min and the detection wavelength was 228 nm. The concentration of glibenclamide in placental perfusate was determined by liquid-liquid extraction pretreatment. Results The linear range of glibenclamide was 2.0 ~ 25.0 μg / mL. The linear equation was y = 0.226x + 0.002, r = 0.999 9 (n = 6) and the relative standard deviation (RSD) was <3.1% RSD <9.5%, methodology recovery was 95.32% ~ 103.35%. Conclusion The HPLC method is sensitive and simple and can be used for the determination of glibenclamide in placental perfusate.