液相色谱-同位素稀释质谱法准确测定人血清中葡萄糖含量

Accurate Determination of Serum Glucose by Liquid Chromatography-Isotope Dilution Mass Spectrometry

以D-[13C6]葡萄糖标记物作为内标,用高效液相色谱-串联质谱法测定血清中的葡萄糖含量。向室温平衡后的血清样品中加入葡萄糖标记物,用乙醇沉淀蛋白,低温离心去除蛋白,上清液过0.22μm有机滤膜。采用LC/MS电喷雾离子源（ESI）,正离子模式,选择离子监测模式进行检测。方法的线性相关系数R2=0.999 5,加标回收率为100.1%~102.8%,检出限为8μg/kg,相对标准偏差0.49%。该方法的样品前处理简单,测定准确高、精密度好,可用于血清中葡萄糖含量的高准确度定值。

A high performance liquid chromatography tandem mass spectrometric method was established for the determination of glucose concentration in serum with the aid of D-glucose internal standard.D-glucose was added to the serum sample in balance at room temperature.Then the protein was precipitated and extracted in anhydrous ethanol with centrifugation at low temperature.The supernatant was filtered through a 0.22 μm syringe filter into HPLC vials.Identification was achieved by electro-spray ionization（ESI） in positive mode using SIM mode.The calibration curve of glucose showed good linearity with correlation coefficient of R2=0.999 5,recoveries of spiked samples were in the range 100.1% to 102.8%,the detection limit of the method was 8 μg/kg（S/N=3）,and the relative standard derivations were 0.49%.The established method is simple,accurate and precise,is appropriate for the determination of serum glucose with great accuracy.