Abstract: To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS) for the determination of urinary catecholamines, including epinephrine(E), norepinephrine(NE) and dopamine(DA). E, NE and DA were separated by WATERS ACQUITY UPLC^® HSS T3 (2.1 mm×100 mm,1.8 μm column) using E-d3, NE-d6 and DA-d4 as internal standards. The mobile phase consisted of 2‰ formic acid aqueous solution (A1) and methyl alcohol (B1, purity=99.9%). The flow rate of 0.50 mL/min was applied to the chromatographic column. Flow starting was performed with A1∶B1=95∶5. Electrospray ionization (ESI) was operated in positive ion mode. The LC-MS/MS was used, with linearity, accuracy (recovery rate), imprecision and lower limit of determination being evaluated. A total of 65 healthy subjects were enrolled, and the reference ranges for E, NE and DA were established. The retention times of E, NE and DA were 0.58, 0.89, 0.63 min, respectively. The determination range was from 0.50 ng/mL to 800.00 ng/mL. The mean recovery ranges were 97.95%, deter mination range 97.78% and 101.03%. The lower limits of determination for E, NE and DA were 0.25, 2.50 and 2.50 ng/mL. The reference ranges were 5.4-25.8 μg/24 h for E, 28.5-73.1 μg/24 h for NE and 269.1-420.5 μg/24 h for DA. The LC-MS/MS for the simultaneous determination of E, NE and DA has been established and is suitable for clinical application. It can provide reliable information for the diagnosis of phaeochromocytoma.

Key words: Epinephrine, Norepinephrine, Dopamine, Urine, Liquid chromatography-tandem mass spectrometry