Simultaneous measurement of urinary metanephrines and catecholamines by liquid chromatography with tandem mass spectrometric detection

This version was published on 1 March 2009

Ann Clin Biochem 2009;46:129-136
© 2009 Association for Clinical Biochemistry


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Original Articles

M J Whiting

Clinical Biochemistry and Pharmacology Laboratory, SA Pathology, Flinders Medical Centre, Bedford Park 5042, South Australia

Corresponding author: Dr Malcolm J Whiting. Email: Malcolm.Whiting{at}

Background: The measurement of catecholamines and metanephrines in urineis an important diagnostic test in biochemical screening forphaeochromocytoma. Tandem mass spectrometry (MSMS) has the potentialto be used in a profiling method for simultaneous assay of theseanalytes.

Methods: Optimal conditions were established for the MSMS detection ofcatecholamines (noradrenalin, adrenalin and dopamine) and metanephrines(normetanephrine and metanephrine), including commercially availableisotopically labelled compounds for use as internal standards.Chromatographic separation of all five polar biogenic amineswas achieved under solvent conditions that were compatible withMSMS and multiple reaction monitoring. Several types of solid-phaseextraction cartridge were used to investigate clean-up conditionsfor urine, and acid-hydrolysates of urine, prior to LC-MSMS.

Results: Total catecholamines and metanephrines from acid-hydrolysedurines, or free catecholamines and free metanephrines from nativeurines, were complexed with diphenyl-boronate and recoveredin high yield from polymer cartridges after elution with formicacid. Direct injection of eluates into the LC-MSMS system allowedquantitation of catecholamines and metanephrines with a runtime of 6 min per sample. Biogenic amine concentrations forpatient urines and quality assurance programme samples, andassay imprecision, were similar to values obtained with high-performanceliquid chromatography methods, which used electrochemical detection.In normal urines, the ratio of free to total catecholamineswas around three-fold higher than the ratio of free to totalmetanephrines.

Conclusion: The assay of urinary catecholamines and metanephrines can beachieved simultaneously using one LC-MSMS method, which is rapidand reduces labour and consumable costs for routine application.

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