Simultaneous Quantification of Plasma Catecholamines and Metanephrines by LC‑MS/MS

Abstract

The quantification of the low levels of catecholamines and metanephrines in biological fluids is important for clinical screening of pheochromocytoma/paraganglioma and diagnosis of overtraining syndrome in athletes. We introduce a novel, accurate and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of these biogenic amines in human plasma. Simple protein precipitation combined with rapid 2-aminoethyl diphenylborinateassisted liquid-liquid extraction allow us to quantify catecholamines and metanephrines over broad concentration ranges. Target analytes were monitored in positive electrospray ionization mode by multiple reaction monitoring. Method performance was validated for linearity, lower limit of quantification, limit of detection, intra-day and inter-assay precision, carry-over, recovery, and matrix effect. The assay was linear within analytical range 25-1000 pg mL −1 for epinephrine, 30-2500 pg mL −1 for norepinephrine, 15-1000 pg mL −1 for dopamine, 25-2000 pg mL −1 for metanephrine and 50-10000 pg mL −1 for normetanephrine, with lower limits of quantification of 15, 20, 10, 15 and 30 pg mL −1 , respectively. The intra-and inter-day precisions for all compounds ranged from 0.4 to 6.9% and from 0.9 to 6.6%, respectively. The efficiency of novel method was confirmed by assaying external quality control samples which demonstrated consistent and accurate results.

A LC–MS/MS method for the simultaneous determination of 6‐cyanodopamine, 6‐nitrodopamine, 6‐nitrodopa, 6‐nitroadrenaline and 6‐bromodopamine in human plasma and its clinical application in patients with chronic kidney disease

“…Deuterated stable isotopes of the analytes are often used as internal standards because they have a mass shift of 1 m / z for each deuterium that substitutes the hydrogen in the structure of a compound. A concern with multiply deuterated internal standards is that they gradually lose the deuterium to hydrogen ions, depending on the solvent that constitutes the standard (Davison et al, 2013), but at the same time a far shift in mass is helpful to eliminate the interference of isotopic precursor ions on the signal of analytes (Davison et al, 2013; Dikunets et al, 2020). A recommendation is to dilute the internal standard only in previously investigated solvents, which will avoid loss of sensitivity of the internal standard (Davison et al, 2013).…”

“…In addition to these detectors, tandem mass spectrometry coupled to HPLC is a robust option to determine the catecholamine content in real samples, as the mass spectrometry (MS) detector has a high selectivity that allows analytes and matrix interferents to be distinguished based on their masses (Lefeuvre et al, 2021). Extraction protocols used in these methods vary from solid‐phase extraction (SPE) on a cation‐exchange column (Takezawa et al, 2000), C 18 cartridges (Talwar et al, 2002) and aluminum oxide columns (He et al, 1997; Li et al, 2000) to liquid–liquid extraction (Dikunets et al, 2020). A different approach than separation‐based methods is batch methods, such as enzyme‐linked immunosorbent assay (ELISA), which have lower selectivity but offer a sensitive, fast and easy‐to‐perform analysis of catecholamines (Kim et al, 2008).…”

Active Inference Theory: The Effect of Sport Training on Tracking Movements Control

“…athlete's perception on the basis of the previous experience, at perf = at perf -at targ , nt perf = nt perf /nt targ ;  execution error at targ = at perf -at plan . To quantify dopamine in the athlete's blood we used the method of high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) [16].…”