Analysis of Catecholamines in Urine by Positive-Ion Electrospray Tandem Mass Spectrometry

Abstract

Abstract Background: Determination of urinary catecholamines (CATs) is considered important for clinical diagnosis of pheochromocytoma, paraganglioma, and neuroblastoma. The major disadvantages of existing tests include relatively long instrumental analysis time and potential interference from drugs and drug metabolites that are structurally similar to CATs. Methods: CATs were extracted from a 300-μL aliquot of urine by a two-step liquid-liquid extraction method specific for compounds containing a catechol group. Chromatographic separation did not require the use of ion-pairing reagents, which typically hinder MS detection but are frequently used in HPLC analysis of CATs. Instrumental analysis was performed by electrospray ionization tandem mass spectrometry (ESI-MS/MS) in the multiple-reaction monitoring mode. Stable-isotope-labeled CATs were used as internal standards. Results: Epinephrine (E), norepinephrine (NE), and dopamine (D) were measured within 3.5 min instrumental run time. Quantification limits were 2.5 μg/L for E and D and 10 μg/L for NE. The total imprecision (CV) was ≤9.6%; extraction recoveries were 71% ± 12%. Conclusions: HPLC with ESI-MS/MS in combination with sample preparation specific to catechol group-containing compounds allows rapid testing for disorders associated with increased CAT concentrations. The method is free of interferences from drugs and drug metabolites, which commonly interfere with HPLC methods.