An isotope dilution capillary electrophoresis/tandem mass spectrometry (CE-MS/MS) method for the simultaneous measurement of choline, betaine, and dimethylglycine concentrations in human plasma

Mauro Forteschi, Angelo Zinellu, Stefano Assaretti, Arduino Mangoni, Gianfranco Pintus, Ciriaco Carru, Salvatore Sotgia

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    7 Citations (Scopus)

    Abstract

    Plasma concentrations of choline, betaine, and dimethylglycine provide valuable information on the flow of methyl groups in key biological processes, particularly during folate deficiency states. We developed a new method to simultaneously measure these analytes in human plasma. Following sample deproteinization using acetonitrile, an aliquot was evaporated to dryness under vacuum to be then taken up by water. Finally, analytes were separated by capillary electrophoresis and detected by electrospray ionization triple-quadrupole mass spectrometry, in multiple reaction monitoring mode, using two stable isotope-labeled internal standards. Linearity of the calibration curves of each analyte was good (R(2) > 0.99). Average limits of detection (LODs) and limits of quantification (LOQs) for choline, betaine, and dimethylglycine were, respectively, 0.43, 0.62, and 0.31 μmol/L and 1.52, 2.11, and 0.97 μmol/L. Mean recovery of three replicates of two spiked concentrations levels was close to 100 % for all of the analytes. Repeatability and intermediate precision, expressed as %RSD of measurements, were <9 %. The method, applied to measure analytes in samples from 30 patients with chronic kidney disease and 30 age- and sex-matched healthy controls, was able to detect differences between groups and the sexes.

    Original languageEnglish
    Pages (from-to)7505-7512
    Number of pages8
    JournalAnalytical and Bioanalytical Chemistry
    Volume408
    Issue number26
    Early online date2016
    DOIs
    Publication statusPublished - 1 Oct 2016

    Keywords

    • Betaine-homocysteine methyltransferase
    • Capillary electrophoresis
    • One-carbon metabolism
    • Quaternary ammonium cations
    • Tandem mass spectrometry

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