同位素标记-超高效液相色谱-串联质谱法快速测定血清25-羟基维生素D3

Rapid determination of serum 25-hydroxyvitamin D3 by isotope labeling-ultra high performance liquid chromatography-tandem mass spectrometry

目的建立同位素标记-超高效液相色谱-串联质谱法快速测定血清25-羟基维生素D 3的方法。方法样品经乙腈提取,饱和硫酸锌沉淀蛋白,然后用正己烷萃取,氮气吹干后85%甲醇定容上样,以0.1%甲醇水-0.1%甲酸水为流动相,经ACQUITY BEH C 18(1.7μm,2.1 mm×100 mm)色谱柱分离,采用多重反应监测正离子模式进行检测。结果该方法在1~100 ng/L内线性关系良好;检出限0.3ng/kg;3个水平加标回收率为92.9%~98.9%,日内相对标准偏差为3.2%~4.2%,日间相对标准偏差为4.4%~4.9%。结论该方法操作简便、灵敏度高,适用于对血清25-羟基维生素D 3的快速检测。

Objective To establish the method for rapidly determining serum 25-hydroxyvitamin D 3 by isotope labeling ultra performance liquid chromatography electrospray ionization tandem mass spectrometry(UPLC-ESI-MS/MS).Methods The samples were extracted with acetonitrile,the protein was precipitated by saturated zinc sulfate,then extracted with n-hexane,after drying with nitrogen,and sampled with 85%methanol at constant volume.The samples were separated by ACQUITY BEH C18(1.7μm,2.1 mm×100 mm)column using 0.1%methanol water-0.1%formic acid water as the mobile phase.The samples were detected by multiple reaction monitoring(MRM)positive ion mode.Results The method had a good linear relationship in the range of 1-100 ng/L,the detection limit(LOD)was 0.3 ng/kg,the recovery rates of three levels were 92.9%-98.9%,the intraday relative standard deviation(RSD)was 3.2%-4.2%and the interday RSD was 4.4%-4.9%.Conclusion This method is simple to operate,highly sensitive and suitable for rapid detection of serum 25-hydroxyvitamin D 3.