Quantification of Methylmalonic Acid in Human Plasma with Hydrophilic Interaction Liquid Chromatography Separation and Mass Spectrometric Detection

Abstract

BACKGROUND: Measurement of methylmalonic acid (MMA) in serum or plasma is useful for diagnosing cobalamin deficiency. We developed a method for quantifying MMA in plasma based on hydrophilic interaction liquid chromatography (HILIC) and single-stage negative electrospray ionization (ESI) mass spectrometry.

Highly sensitive and selective measurement of underivatized methylmalonic acid in serum and plasma by liquid chromatography-tandem mass spectrometry

“…A variety of methods have been reported for the measurement of plasma MMA, including capillary electrophoresis [5], high-performance liquid chromatography (HPLC) [6], gas chromatography-mass spectrometry (GC-MS) [7], and liquid chromatography-tandem mass spectrometry (LC-MS/ MS) [8]. The vitamin B12 experts from the National Health and Nutrition Examination Survey (NHANES) encourage the use of LC-MS/MS for the analysis of plasma MMA [9].…”

“…The measurement of plasma MMA is confounded by its hydrophilic and non-volatile nature, low molecular weight (118 Da), and isobaric interference from succinic acid (SA) ( Fig. 1) which can have circulating concentrations 100-fold higher than MMA [8,10]. Several LC-MS/MS methods for the measurement of plasma MMA have been developed [8,[11][12][13][14][15][16].…”

“…1) which can have circulating concentrations 100-fold higher than MMA [8,10]. Several LC-MS/MS methods for the measurement of plasma MMA have been developed [8,[11][12][13][14][15][16]. However, many of these methods have limitations including tedious sample preparation involving derivatization, lengthy chromatography, or insufficient sensitivity and/ or specificity.…”

Semi-automated quantification of methylmalonic acid in human serum by LC-MS/MS

“…Although LC-MS/MS is rapidly replacing GC-MS in clinical laboratories due to its relative methodological simplicity [13][14][15][16][17] most current LC-MS/MS methods for MMA determination still use derivatization [13][14][15][16] and few utilize automation. Our goal was to simplify and automate preanalytical handling and to speed separation to increase throughput, while retaining accuracy and precision in a routine clinical setting.…”

An on-line solid phase extraction procedure for the routine quantification of urinary methylmalonic acid by liquid chromatography-tandem mass spectrometry

“…Apart from several articles which describe analysis of underivatized MMA by LC-MS/MS using electrospray ionisation in the negative mode (12,13), the majority of published work uses the positive mode. This may be attributed to a rather low ion yield, using negative ionization in many instruments including ours.…”