Determination of homovanillic acid in urine by stable isotope dilution and electrospray tandem mass spectrometry

Mark J. Magera, Amy L. Stoor, Janice K. Helgeson, Dietrich Matern, Piero Rinaldo

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Abstract

We have developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the analysis of homovanillic acid (HVA), a biochemical marker for catecholamine and neurotransmitter metabolism. Urine specimens are spiked with 5 μg of a stable-isotope labeled internal standard, 13C618O-HVA, and prepared by automated solid phase extraction. Residues were dissolved in acetonitrile: 0.05% aqueous acetic acid and analyzed by MS/MS in the selected reaction monitoring mode (HVA: m/z 181 to m/z 137; 13C618O-HVA: m/z 189 to m/z 145) after separation using a Discovery RP Amide C16 column. Consecutive calibrations (n = 7) between 0.52 and 16.7 mg/l exhibited consistent linearity and reproducibility. At a urine concentration of 0.51 mg/l, the signal-to-noise ratio for HVA was 21:1. Inter- and intra-assay CVs ranged from 0.3% to 11.4%, at mean concentrations ranging 1.8 to 22.7 mg/l. Recovery of HVA added to urine ranged between 94.7% and 105% (1.25 mg/l added), 92.0% and 102% (5.0 mg/l), and 96.0% and 104% (10 mg/l). LC-MS/MS is well suited to replace an HPLC method for routine HVA determination, by providing positive identification, faster turn around time, virtually no repeat analyses and a 44% reduction of personnel necessary to perform the testing.

Original languageEnglish (US)
Pages (from-to)35-41
Number of pages7
JournalClinica Chimica Acta
Volume306
Issue number1-2
DOIs
StatePublished - 2001

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Keywords

  • Electrospray ionization
  • Homovanillic acid
  • Stable isotope dilution
  • Tandem mass spectrometry

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

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