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Integration of Miniaturized Solid Phase Extraction and LC-MS/MS Detection of 3-Nitrotyrosine in Human Urine for Clinical Applications
Published on: July 14, 2017
A simple liquid chromatography-tandem mass spectrometry method for measuring metanephrine and normetanephrine in
Jessica Gabler1, Abby Miller, Sihe Wang
1Department of Clinical Pathology, Cleveland Clinic, Cleveland, OH, USA.
Background:
Measuring urinary fractionated metanephrines is one of the initial tests in the diagnosis of pheochromocytoma. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) represents the most specific and accurate technology for this purpose. The goal of this work was to develop a simple LC-MS/MS method for measuring metanephrines in urine.
Methods:
Each urine sample was complexed with diphenylboronic acid, and purified on a Bond-Elute Plexa cartridge. The extract was concentrated and analyzed on a short Atlantis T3 column in 8.5 min. Metanephrines and their deuterated internal standards were monitored in positive electrospray ionization mode by multiple reaction monitoring.
Results:
Ion suppression was observed, but was compensated for by the respective internal standard. The analytical measurement range was 0.2-27.4 μmol/L and 0.3-14.6 μmol/L for metanephrine and normetanephrine, respectively. The intra-assay and total coefficient of variation throughout the linear ranges was 2.03%-2.11% and 2.20%-3.80% for metanephrine, and 4.50%-8.09% and 9.00%-10.00% for normetanephrine, respectively. Comparison with a commercial HPLC method using patient samples (n=65) by Passing-Bablok regression showed a slope of 1.000 and 1.014, y-intercept of -0.080 and -0.067, a correlation coefficient of 0.8830 and 0.9022, and a mean difference of 14.0% and -0.43% for metanephrine and normetanephrine, respectively.
Conclusions:
This simple method for urine metanephrines is suitable for clinical use.
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