Validation of a Sensitive, Simple and High-Throughput UPLC-MS/MS Method for Quantification of Catecholamines and Their Metabolites in Serum and Urine: Application in Clinical Analysis.

This article focuses on the development and validation of a sensitive, simple, and high-throughput ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneous quantification of catecholamines (CAs)—dopamine (DA), epinephrine (E), norepinephrine (NE)—and their metabolites metanephrine (MN), normetanephrine (NMN), and 3-methoxytyramine (3-MT) in human serum and urine. The method employs a 96-well solid-phase extraction (SPE) cartridge and a pentafluorophenyl (PFP) column, achieving a short run time of 4 minutes without requiring derivatization, and demonstrates excellent linearity, accuracy, precision, recovery, and stability. For the first time, the study compares analyte levels between plasma and serum, finding significant correlations but also significant differences, underscoring the need for serum-specific reference intervals in clinical diagnostics. The validated method offers a practical tool for clinical and research applications involving CA measurement in multiple biological matrices.