Abstract
Plasma metanephrines are measured to aid in the diagnosis of pheochromocytomas. In patients with pheochromocytomas there is excessive production of catecholamines and metanephrines. Measurement of plasma free metanephrines is one of the first-line clinical tests that are used for the diagnosis and follow-up of pheochromocytoma. We describe here a liquid chromatography-tandem mass spectrometry method to measure free metanephrines in plasma. Free metanephrine and normetanephrine are extracted via solid-phase extraction. After extraction and evaporation, the reconstituted supernatant is analyzed by high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). The MS/MS is set to selective reaction monitoring mode (180.1 → 148.1 m/z for metanephrine, 183.1 → 168.1 for d3-metanephrine, 166.1 → 134.1 m/z for normetanephrine, and 169.1 → 137.2 m/z for d3-normetanephrine) with positive electrospray ionization. Quantitation is based on peak area ratio of the analyte to its respective deuterated internal standard. The assay is linear from 5.9 to 4090.0 pg/mL for metanephrine and 22.0 to 4386.7 pg/mL for normetanephrine with precision of <6 % over the ranges.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lenders JW, Pacak K, Walther MM, Linehan WM, Mannelli M, Friberg P, Keiser HR, Goldstein DS, Eisenhofer G (2002) Biochemical diagnosis of pheochromocytoma: which test is best? JAMA 287:1427–1434
Grouzmann E, Drouard-Troalen L, Baudin E, Plouin PF, Muller B, Grand D, Buclin T (2010) Diagnostic accuracy of free and total metanephrines in plasma and fractionated metanephrines in urine of patients with pheochromocytoma. Eur J Endocrinol 162:951–960
Pacak K, Eisenhofer G, Ahlman H, Bornstein SR, Gimenez-Roqueplo AP, Grossman AB, Kimura N, Mannelli M, McNicol AM, Tischler AS (2007) International symposium on pheochromocytoma. Pheochromocytoma: recommendations for clinical practice from the First International Symposium October 2005. Nat Clin Pract Endocrinol Metab 3:92–102
Pillai D, Callen S (2010) Pilot quality assurance programme for plasma metanephrines. Ann Clin Biochem 47:137–142
Singh RJ, Eisenhofer G (2007) High-throughput, automated, and accurate biochemical screening for pheochromocytoma: are we there yet? Clin Chem 53:1565–1567
Peaston RT, Graham KS, Chambers E, van der Molen JC, Ball S (2010) Performance of plasma free metanephrines measured by liquid chromatography-tandem mass spectrometry in the diagnosis of pheochromocytoma. Clin Chim Acta 411:546–552
de Jong WH, Graham KS, van der Molen JC, Links TP, Morris MR, Ross HA, de Vries EG, Kema IP (2007) Plasma free metanephrine measurement using automated online solid-phase extraction HPLC tandem mass spectrometry. Clin Chem 53:1684–1693
Lagerstedt SA, O’Kane DJ, Singh RJ (2004) Measurement of plasma free metanephrine and normetanephrine by liquid chromatography-tandem mass spectrometry for diagnosis of pheochromocytoma. Clin Chem 50:603–611
Gabler J, Yuan C, Woroniecki W, Liu H, Wang S (2012) A Sensitive and Interference-Free Liquid Chromatography Tandem Mass Spectrometry Method for Measuring Metanephrines in Plasma. J Chromatograph Separat Techniq 4:195. doi:10.4172/2157-7064.1000195
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Heideloff, C., Payto, D., Wang, S. (2016). Quantitation of Free Metanephrines in Plasma by Liquid Chromatography-Tandem Mass Spectrometry. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Biomolecular Analysis. Methods in Molecular Biology, vol 1378. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3182-8_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3182-8_16
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3181-1
Online ISBN: 978-1-4939-3182-8
eBook Packages: Springer Protocols