Tecleab A G, Schofield R C, Ramanathan L V, Carlow Dean C
Department of Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA.
J Chromatogr Sep Tech. 2016 Oct;7(5). doi: 10.4172/2157-7064.1000336. Epub 2016 Sep 21.
A simple and sensitive method for the detection of methylmalonic acid in serum without derivatization has been developed. This method implements protein precipitation using methanol followed by additional sample clean up by turbulent flow liquid chromatography (TFLC). The sample was directly injected into the turbulent flow liquid chromatography tandem mass spectrometry system (TFLC-MS/MS) for online extraction followed by HPLC separation. The eluent was transferred to the mass spectrometer and ionized by heated electrospray negative ionization (HESI) and the analyte was quantified using a six-point calibration curve. The validated analytical measurement range (AMR) is 30-1,000 nMol/L. Dilutions of 10 and 200-fold were validated giving a clinical reportable range (CRR) of 30-200,000 nMol/L. The between-day and within-day imprecision values at concentrations spanning the AMR were less than 15%. This method was compared to an established LC-MS/MS method at a CLIA certified national reference laboratory and shows an excellent correlation with our TFLC-MS/MS method.
已开发出一种无需衍生化即可检测血清中甲基丙二酸的简单且灵敏的方法。该方法先用甲醇进行蛋白质沉淀，然后通过湍流液相色谱（TFLC）对样品进行进一步净化。将样品直接注入湍流液相色谱串联质谱系统（TFLC-MS/MS）进行在线萃取，随后进行高效液相色谱（HPLC）分离。洗脱液转移至质谱仪，通过加热电喷雾负离子化（HESI）进行离子化，并使用六点校准曲线对分析物进行定量。验证后的分析测量范围（AMR）为30 - 1000 nMol/L。对10倍和200倍稀释液进行了验证，得出临床报告范围（CRR）为30 - 200,000 nMol/L。在跨越AMR的浓度下，日间和日内不精密度值均小于15%。在一家经CLIA认证的国家参考实验室，将该方法与一种既定的液相色谱-串联质谱（LC-MS/MS）方法进行了比较，结果表明其与我们的TFLC-MS/MS方法具有良好的相关性。
