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Quantitation of Free Metanephrines in Plasma by Liquid Chromatography-Tandem Mass Spectrometry

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Clinical Applications of Mass Spectrometry in Biomolecular Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1378))

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.

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References

  1. 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

    Article  CAS  PubMed  Google Scholar 

  2. 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

    Article  CAS  PubMed  Google Scholar 

  3. 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

    Article  PubMed  Google Scholar 

  4. Pillai D, Callen S (2010) Pilot quality assurance programme for plasma metanephrines. Ann Clin Biochem 47:137–142

    Article  CAS  PubMed  Google Scholar 

  5. Singh RJ, Eisenhofer G (2007) High-throughput, automated, and accurate biochemical screening for pheochromocytoma: are we there yet? Clin Chem 53:1565–1567

    Article  CAS  PubMed  Google Scholar 

  6. 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

    Article  CAS  PubMed  Google Scholar 

  7. 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

    Article  PubMed  Google Scholar 

  8. 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

    Article  CAS  PubMed  Google Scholar 

  9. 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

    Google Scholar 

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Correspondence to Sihe Wang .

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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

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  • 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

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