Abstract
Abstract
Background: Widely used HPLC methods for quantification of metanephrine and normetanephrine in urine often have long analysis times and are frequently plagued by drug interferences. We describe a gas chromatography-mass spectrometry method designed to overcome these limitations.
Methods: Metanephrine and normetanephrine conjugates were converted to unconjugated metanephrine and normetanephrine by acid hydrolysis. To avoid the rapid decomposition of the deuterated internal standards (metanephrine-d3 and normetanephrine-d3) under hydrolysis conditions, the internal standards were added after hydrolysis. Solid-phase extraction was used to isolate the hydrolyzed metanephrines from urine. Samples were concentrated by evaporation, then derivatized simultaneously with N-methyl-N-(trimethylsilyl)trifluoroacetamide and N-methyl-bis-heptafluoro-butryamide at room temperature.
Results: The assay was linear from 25 to 7000 μg/L. The intraassay CVs were <5% and the interassay CVs <12%. Comparison with a routine HPLC method (n = 192) by Deming regression yielded a slope of 1.00 ± 0.02 μg/L, an intercept of −5.8 ± 7.8 μg/L, and Sy|x = 50.6 μg/L for metanephrine and a slope of 0.94 ± 0.03, intercept of 19 ± 11 μg/L, and Sy|x = 60 μg/L for normetanephrine. The correlation coefficients (r) were calculated after log transformation of the data and gave r = 0.97 for metanephrine and r = 0.97 for normetanephrine. Interference from common medications or drug metabolites was seen in <1% of samples. The time between sequential injections was <7 min.
Conclusions: This new gas chromatography-mass spectrometry assay for total fractionated metanephrines is rapid, compares well with a standard HPLC assay, and avoids most drug interferences that commonly affect HPLC assays for urine metanephrines.