使用液相色谱-串联质谱法对血浆中的甲基丙二酸进行定量分析。
Quantitation of methylmalonic acid in plasma using liquid chromatography-tandem mass spectrometry.
作者信息
Fasching Claudine, Singh Jasbir
机构信息
Pathology and Laboratory Medicine Service, Veterans Affairs Medical Center, Minneapolis, MN, USA.
出版信息
Methods Mol Biol. 2010;603:371-8. doi: 10.1007/978-1-60761-459-3_36.
Methylmalonic acid (MMA), a biochemical marker for vitamin B12 deficiency, is commonly measured in serum and urine using liquid chromatography-tandem mass spectrometry (LC-MS-MS) with electrospray ionization. Improvements in sample preparation and the ability to analyze MMA without derivatization have further simplified the analysis. In the method we describe, 200 microl of plasma spiked with deuterated d3-MMA internal standard is deproteinized using ultrafiltration. After acidification, the ultrafiltrate is injected into the LC-MS-MS system and MMA eluted under isocratic conditions. MMA and d3-MMA are monitored in multiple reaction monitoring (MRM) in the negative ion mode.
甲基丙二酸（MMA）是维生素B12缺乏的生化标志物，通常采用电喷雾电离的液相色谱-串联质谱法（LC-MS-MS）测定血清和尿液中的MMA。样品制备的改进以及无需衍生化即可分析MMA的能力进一步简化了分析过程。在我们所描述的方法中，向200微升添加了氘代d3-MMA内标的血浆中加入超滤法进行去蛋白处理。酸化后，将超滤液注入LC-MS-MS系统，MMA在等度条件下洗脱。在负离子模式下，通过多反应监测（MRM）对MMA和d3-MMA进行监测。
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