A novel method for the measurement of plasma metanephrines using online solid phase extraction-liquid chromatography tandem mass spectrometry

Ann Clin Biochem. 2015 May;52(Pt 3):361-9. doi: 10.1177/0004563214546560. Epub 2014 Jul 21.

Abstract

Background: Measurement of plasma metanephrine, normetanephrine and 3-methoxytyramine is useful in the diagnosis of phaeochromocytomas, but many assays require a large volume of plasma due to poor assay sensitivity, and often require lengthy sample preparation. Our aim was to develop a method for measurement of plasma metanephrines using a small sample volume with minimal hands-on preparation.

Methods: Samples were deproteinised using 10 K spin filters prior to online solid phase extraction using a Waters Acquity UPLC Online SPE Manager (Waters, Manchester, UK) coupled to a Waters Xevo TQ-S mass spectrometer (Waters, Manchester, UK). The assay was validated and results compared to a previously published method.

Results: We achieved a limit of quantification of 37.5 pmol/L for metanephrine and 3-methoxytyramine and 75 pmol/L for normetanephrine using only 150 µL of sample. The assay was linear up to 30,000 pmol/L for all analytes and in a method comparison study results showed good agreement with a previously published LC-MS/MS assay.

Conclusions: We have developed a simple method for measurement of plasma metanephrine, normetanephrine and 3-methoxytyramine using only 150 µL of sample. There is minimal hands-on sample preparation required and the assay is suitable for routine use in a clinical laboratory.

Keywords: 3-methoxytyramine; LC-MS/MS; Metanephrine; normetanephrine; online solid phase extraction; phaeochromocytoma.

MeSH terms

  • Chromatography, Liquid / methods
  • Humans
  • Metanephrine / blood*
  • Solid Phase Extraction / methods*
  • Tandem Mass Spectrometry / methods*

Substances

  • Metanephrine