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Received on July 18, 2003
Accepted on December 19, 2003
Endocrinology and Metabolism |
1 Department of Laboratory Medicine & Pathology, Mayo Clinic and Foundation, 200 First Street SW, Rochester, MN 55905
* To whom correspondence should be addressed. E-mail: singh.ravinder{at}mayo.edu.
Background: Quantification of plasma free metanephrines is usually accomplished by HPLC with electrochemical detection, but sample preparation is labor-intensive and time-consuming, run times are long, and interfering substances sometimes obscure the relevant peaks. The aim of this study was to develop a sensitive and specific LC-MS/MS method for plasma free metanephrines.
Methods: After solid-phase extraction, chromatographic separation of normetanephrine (NMN) and metanephrine (MN) was accomplished by use of a cyano analytical column. NMN, MN, d3-NMN, and d3-MN positive ions were detected in the multiple-reaction monitoring mode using the specific transitions m/z 166
134, 180148, 169137, and 183151, respectively, with an atmospheric pressure chemical ionization source.
Results: Multiple calibration curves exhibited consistent linearity and reproducibility. Interassay imprecision values (CV; n = 20) for NMN at 0.64, 1.9, and 2.7 nmol/L were 6.6%, 7.8%, and 13%, respectively. Interassay CV for MN at 0.60, 1.2, and 2.1 nmol/L (n = 20) were 9.2%, 6.8%, and 9.8%, respectively. The mean recoveries of NMN and MN relative to the internal standard were 100% and 96%, respectively. The assays were linear between 0.20 and 10.0 nmol/L. Deming regression of HPLC and LC-MS/MS results yielded slopes of 0.93 (95% confidence interval, 0.89-0.98) and 0.89 (0.85-0.93) and y-intercepts of -0.16 and 0.03 nmol/L for NMN (n = 132) and MN (n = 92), respectively.
Conclusions: This novel LC-MS/MS method provides a precise, rapid, and specific alternative to HPLC for the quantification of the low nanomolar concentrations of free metanephrines in plasma.
The following articles in journals at HighWire Press have cited this article:
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R. J. Singh and G. Eisenhofer High-Throughput, Automated, and Accurate Biochemical Screening for Pheochromocytoma: Are We There Yet? Clin. Chem., September 1, 2007; 53(9): 1565 - 1567. [Full Text] [PDF]
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R. L. Taylor, S. K. Grebe, and R. J. Singh Quantitative, Highly Sensitive Liquid Chromatography-Tandem Mass Spectrometry Method for Detection of Synthetic Corticosteroids Clin. Chem., December 1, 2004; 50(12): 2345 - 2352. [Abstract] [Full Text] [PDF]
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