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Development and validation of a specific and sensitive LC-MS/MS method for quantification of urinary catecholamines and application in biological variation studies

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Abstract

Catecholamines are a class of biogenic amines that play an important role as neurotransmitters and hormones. We developed and validated a rapid, specific and sensitive LC-MS/MS method for quantitative determination of catecholamines in human urine. Linearity, specificity, sensitivity, precision, accuracy, matrix effect, carryover, analyte stability, method comparison and reference range were evaluated. The catecholamine measurements were not affected by 35 structurally-related drugs and metabolites. The outstanding specificity was achieved by use of a specific diphenylborate-based solid phase extraction and subsequent selective LC-MS/MS analysis. Excellent sensitivity, accuracy and precision (average intra-assay variations <2.9 % and inter-assay variations <4.6 %) were obtained. The method was successfully applied in the study of day-to-day biological within- and between-subject variations of 25 healthy people under free-living conditions over three consecutive days. We observed that catecholamine excretions for second morning sampling had least day-to-day within-subject variation and excellent reproducibility. This work is one of the rare studies on these topics and represents the first utilization of advanced LC-MS/MS technology. Additionally, we found significant correlations between spot and conventional 24 h collections of human urine (n = 22, r > 0.853, p < 0.0001). These findings suggest that determining the catecholamine concentrations in the second morning urine sample presents accurate, convenient and reliable measurement of catecholamine excretions. In addition, consistent and significant diurnal variations for norepinephrine and epinephrine excretions were observed during the three-day period, while dopamine did not exhibit a diurnal rhythm. The LC-MS/MS method presented here is rapid, sensitive and specific, which could be an advantage in clinical laboratories.

Diurnal variation of urinary catecholamines for 25 healthy people in three consecutive days

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References

  1. Rosano TG, Swift TA, Hayes LW (1991) Advances in catecholamine and metabolite measurements for diagnosis of pheochromocytoma. Clin Chem 37:1854–1867

    CAS  Google Scholar 

  2. Gerlo EAM, Sevens C (1994) Urinary and plasma catecholamines and urinary catecholamine metabolites in pheochromocytoma: diagnostic value in 19 cases. Clin Chem 40:250–256

    CAS  Google Scholar 

  3. Peaston RT, Weinkove C (2004) Measurement of catecholamines and their metabolites. Ann Clin Biochem 41:17–38

    Article  CAS  Google Scholar 

  4. Eisenhofer G, Kopin IJ, Goldstein DS (2004) Catecholamine metabolism: a contemporary view with implications for physiology and medicine. Pharmacol Rev 56:331–349

    Article  CAS  Google Scholar 

  5. Goldstein DS (2010) Catecholamines 101. Clin Auton Res 20:331–352

    Article  Google Scholar 

  6. Lechin F, van der Dijs B (2009) Crosstalk between the autonomic nervous system and the central nervous system: Mechanistic and therapeutic considerations for neuronal, immune, vascular, and somatic based diseases. In: Maiese K (ed) Neurovascular Medicine: Pursuing Cellular Longevity for Healthy Aging. Oxford University Press, New York

    Google Scholar 

  7. Dvoráková M, Jezová D, Blazícek P, Trebatická J, Skodácek I, Suba J, Iveta W, Rohdewald P, Duracková Z (2007) Urinary catecholamines in children with attention deficit hyperactivity disorder (ADHD): modulation by a polyphenolic extract from pine bark pycnogenol). Nutr Neurosci 10:151–157

    Article  Google Scholar 

  8. Pliszka SR, Maas JW, Javors MA, Rogeness GA, Baker J (1994) Urinary catecholamines in attention-deficit hyperactivity disorder with and without comorbid anxiety. J Am Acad Child Adolesc Psychiatry 33:1165–1173

    Article  CAS  Google Scholar 

  9. Pliszka SR, McCracken JT, Maas JW (1996) Catecholamines in attention-deficit hyperactivity disorder: current perspectives. J Am Acad Child Adolesc Psychiatry 35:264–272

    Article  CAS  Google Scholar 

  10. Hughes JW, Watkins L, Blumenthal JA, Kuhn C, Sherwood A (2004) Depression and anxiety symptoms are related to increased 24-hour urinary norepinephrine excretion among healthy middle-aged women. J Psychosom Res 57:353–358

    Article  Google Scholar 

  11. Roy A, Pickar D, Douillet P, Karoum F, Linnoila M (1986) Urinary monoamines and monoamine metabolites in subtypes of unipolar depressive disorder and normal controls. Psychol Med 16:541–546

    Article  CAS  Google Scholar 

  12. Levite M (2012) Nerve-driven Immunity: Neurotransmitters and neuropeptides in the immune system. Springer-Verlag/Wien

  13. Tsunoda M (2006) Recent advances in methods for the analysis of catecholamines and their metabolites. Anal Bioanal Chem 386:506–514

  14. Bicker J, Fortuna A, Alves G, Falcão A (2013) Liquid chromatographic methods for the quantification of catecholamines and their metabolites in several biological samples-a review. Anal Chim Acta 768:12–34

    Article  CAS  Google Scholar 

  15. Fraser CG, Harris EK (1989) Generation and application of data on biological variation in clinical chemistry. Crit Rev Clin Lab Sci 27:409–437

    Article  CAS  Google Scholar 

  16. Ricós C, Jiménez CV, Hernández A, Simón M, Perich C, Alvarez V, Minchinela J, Maciá M (1994) Biological variation in urine samples used for analyte measurements. Clin Chem 40:472–477

    Google Scholar 

  17. Bondanelli M, Ambrosio MR, Franceschetti P, Margutti A, Trasforini G, Degli Uberti EC (1999) Diurnal rhythm of plasma catecholamines in acromegaly. J Clin Endocrinol Metab 84:2458–2467

    CAS  Google Scholar 

  18. Schöfl C, Becker C, Prank K, von zur Mühlen A, Brabant G (1997) Twenty-four-hour rhythms of plasma catecholamines and their relation to cardiovascular parameters in healthy young men. Eur J Endocrinol 137:675–683

    Article  Google Scholar 

  19. Marc DT, Ailts JW, Ailts-Campeau DC, Bull MJ, Olson KL (2011) Neurotransmitters excreted in the urine as biomarkers of nervous system activity: validity and clinical applicability. Neurosci Biobehav Rev 35:635–644

    Article  CAS  Google Scholar 

  20. Fibiger W, Singer G, Miller AJ, Armstrong S, Datar M (1984) Cortisol and catecholamines changes as functions of time-of-day and self-reported mood. Neurosci Biobeh Rev 8:523–530

    Article  CAS  Google Scholar 

  21. Hansen AM, Garde AH, Christensen JM, Eller NH, Netterstrøm B (2001) Reference intervals and variation for urinary epinephrine, norepinephrine and cortisol in healthy men and women in Denmark. Clin Chem Lab Med 39:842–849

    Article  CAS  Google Scholar 

  22. Nichkova M, Wynveen PM, Marc DT, Huisman H, Kellermann GH (2013) Validation of an ELISA for urinary dopamine: applications in monitoring treatment of dopamine-related disorders. J Neurochem 125:724–735

    Article  CAS  Google Scholar 

  23. Curtin F, Walker JP, Shulz P (1996) Day-to-day intraindividual reliability and interindividual differences in monoamines excretion. J Affect Disord 38:173–178

    Article  CAS  Google Scholar 

  24. Souza MCP, Rumpler WV, Douglass LW, Howe JC (1998) Urinary catecholamine excretion in men and women: between- and within-subject variation. J Nutr Biochem 9:396–401

  25. Talwar D, Williamson C, McLaughlin A, Gill A, O’Reilly DS (2002) Extraction and separation of urinary catecholamines as their diphenyl boronate complexes using C18 solid-phase extraction sorbent and high-performance liquid chromatography. J Chromatogr B 769:341–349

    Article  CAS  Google Scholar 

  26. De Jong WHA, De Vries EGE, Kema IP (2011) Current status and future developments of LC-MS/MS in clinical chemistry for quantification of biogenic amines. Clin Biochem 44:95–103

  27. Kushnir MM, Urry FM, Frank EL, Roberts WL, Shushan B (2002) Analysis of catecholamines in urine by positive-ion electrospray tandem mass spectrometry. Clin Chem 48:323–331

  28. Whiting MJ (2009) Simultaneous measurement of urinary metanephrines and catecholamines by liquid chromatography with tandem mass spectrometric detection. Ann Clin Biochem 46:129–136

    Article  CAS  Google Scholar 

  29. De Jong WHA, De Vries EGE, Wolffenbuttel BHR, Kema IP (2010) Automated mass spectrometric analysis of urinary catecholamines using on-line solid phase extraction. J Chromatogr B 878:1506–1512

  30. Peitzsch M, Pelzel D, Glöckner S, Prejbisz A, Fassnacht M, Beuschlein F, Januszewicz A, Siegert G, Eisenhofer G (2013) Simultaneous liquid chromatography tandem mass spectrometric determination of urinary free metanephrines and catecholamines, with comparisons of free and deconjugated metabolites. Clin Chim Acta 418:50–58

    Article  CAS  Google Scholar 

  31. Gosetti F, Mazzucco E, Gennaro MC, Marengo E (2013) Simultaneous determination of sixteen underivatized biogenic amines in human urine by HPLC-MS/MS. Anal Bioanal Chem 405:907–916

    Article  CAS  Google Scholar 

  32. Moriarty M, Lee A, O’Connell B, Kelleher A, Keeley H, Furey A (2011) Development of an LC-MS/MS method for the analysis of serotonin and related compounds in urine and the identification of a potential biomarker for attention deficit hyperactivity/hyperkinetic disorder. Anal Bioanal Chem 401:2481–2493

    Article  CAS  Google Scholar 

  33. Clinical and Laboratory Standards Institute (2007) Mass spectrometry in the clinical laboratory: General principles and guidance; approved guideline. CLSI document C 50-A

  34. Van Eeckhaut A, Lanckmans K, Sarre S, Smolders I, Michotte Y (2009) Validation of bioanalytical LC-MS/MS assays: evaluation of matrix effects. J Chromatogr B 877:2198–2207

    Article  Google Scholar 

  35. White IR, Brunner EJ, Barron JL (1995) A comparison of overnight and 24 hour collection to measure urinary catecholamines. J Clin Epidemiol 48:263–267

    Article  CAS  Google Scholar 

  36. Chan EC, Wee PY, Ho PC (2000) Evaluation of degradation of urinary catecholamines and metanephrines and deconjugation of their sulfoconjugates using stability-indicating reversed-phase ion-pair HPLC with electrochemical detection. J Pharm Biomed Anal 22:515–526

    Article  CAS  Google Scholar 

  37. De Jong WHA, Post WJ, Kerstens MN, De Vries EGE, Kema IP (2010) Elevated urinary free and deconjugated catecholamines after consumption of a catecholamine-rich diet. J Clin Endocrinol Metab 95:2851–2855

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Acknowledgments

We thank Aimee Thiel and Jodi Branstad for their excellent assistance in the clinical studies. We gratefully acknowledge Melissa Ahrens for editing the manuscript.

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Authors and Affiliations

  1. Pharmasan Labs, Inc, 373 280th Street, Osceola, WI, 54020, USA

    Xiaoguang (Sunny) Li, Shu Li, Paul Wynveen, Kathy Mork & Gottfried Kellermann

  2. NeuroScience, Inc, 373 280th Street, Osceola, WI, 54020, USA

    Gottfried Kellermann

Authors
  1. Xiaoguang (Sunny) Li
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  2. Shu Li
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  3. Paul Wynveen
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  4. Kathy Mork
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  5. Gottfried Kellermann
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Correspondence to Xiaoguang (Sunny) Li.

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Li, X., Li, S., Wynveen, P. et al. Development and validation of a specific and sensitive LC-MS/MS method for quantification of urinary catecholamines and application in biological variation studies. Anal Bioanal Chem 406, 7287–7297 (2014). https://doi.org/10.1007/s00216-014-8120-1

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  • DOI: https://doi.org/10.1007/s00216-014-8120-1

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