Methylmalonic acid (MMA) determination in serum or plasma is used as a marker of cobalamin (vitamin B12) deficiency. Separation of MMA by reversed-phase liquid chromatography is difficult due to poor retention. Recently, the combination of hydrophilic interaction liquid chromatography (HILIC) with mass spectrometry has gained interest for separation of polar compounds such as MMA. This study describes fully automated sample preparation and analysis for the quantification of MMA in serum and plasma using HILIC separation with tandem mass spectrometry. Linear calibration, accuracy, precision and recovery were calculated in water, serum and plasma. Correlation coefficients > 0.997 were achieved in all matrices. Precision ranged between 0.8% and 1.1%, accuracy between 99.8% and 101% and recovery between 115% and 118%, all within accepable limits.
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Automating the analysis of Methylmalonic Acid in plasma and serum using a GERSTEL Multi Purpose Sampler coupled to liquid chromatography tandem mass spectrometry detection