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

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Author

  • Dick Nelson
  • Ning Xu
  • Joyce Carlson

Summary, in English

Background. Methylmalonic acid (MMA), a sensitive biomarker of functional vitamin B12 deficiency, is commonly determined by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods using manual extraction and derivatization of MMA to reduce polarity prior to separation. Methods. In the present study we introduce a semi-automated extraction on a strong anion exchanger, HPLC separation on a BEH-amide column to separate serum MMA from its abundant isoform, succinic acid, followed by MS/MS detection and quantification. Results. The extraction of MMA plus internal standard provides full recovery and the method is linear between 0.03 mu mol/L and 20.0 mu mol/L (r(2) = 1.0) with intra-and inter-assay imprecision of 2.2%. Agreement with other laboratories has been demonstrated in external proficiency testing. Compared to both conventional GC-MS and LC-MS/MS methods, the correlation is r(2) > 0.99. Conclusions. The use of robotic pipetting, elimination of derivatization and improved separation by the BEH-amide column combined with HILIC chromatographic conditions significantly improve sample throughput compared to conventional methods. Using a single pipetting robot and LC-MS/MS instrument, this method is currently performing 180 analyses per day from 10 regional hospitals and several additional distant sites.

Publishing year

2012

Language

English

Pages

441-446

Publication/Series

Scandinavian Journal of Clinical & Laboratory Investigation

Volume

72

Issue

6

Document type

Journal article

Publisher

Informa Healthcare

Topic

  • Medicinal Chemistry
  • Pharmacology and Toxicology

Keywords

  • Chromatography
  • high performance liquid
  • tandem mass spectrometry
  • gas
  • chromatography-mass spectrometry
  • methylmalonic acid
  • vitamin B12

Status

Published

ISBN/ISSN/Other

  • ISSN: 1502-7686