Urinary catecholamines and their methylated metabolites are biochemical indicators of pheochromocytoma, paraganglioma and neuroblastoma. A rapid and precise analytical method based on solid-phase extraction (SPE) and liquid chromatography separation coupled to high-resolution mass spectrometry (LC-HRMS) was developed and validated to measure urinary catecholamines (epinephrine (E), norepinephrine (NorE), dopamine (D)) and total methylated metabolites (normetanephrine (NorMN), metanephrine(MN) and 3-methoxytyramine (3-MT)) in a clinical setting. Results of 51 urine specimens measured using this LC-HRMS method were compared with a liquid chromatography assay with electrochemical detection (LC-EC). Urine samples (200 μL) were spiked with an internal standard solution followed by SPE purification. In the case of total methylated metabolites, urine was hydrolyzed before SPE purification. Separation was achieved on an Acclaim Mixed Mode WCX column, with an 8.5 min runtime. All compounds were detected in electrospray positive ionization mode with a parallel reaction monitoring acquisition and quantified with a linear regression (r2>0.998) between 2 and 200 µg/L (10.9-1090; 11.8-1182 nmol/L) for E and NorE respectively and between 10 and 1000 µg/L for others (65.2-6520; 50.7-5070; 54.5-5450 ; 59.8-5980 nmol/L for D, M, NorMN and 3-MT, respectively). Overall imprecision and bias did not exceed 15%. No significant matrix effect was observed. Correlation between the two assays was good except for epinephrine. Epinephrine concentrations measured by LC-EC method were slightly higher than values obtained with LC-HRMS method but without impact on clinical decision. This LC-HRMS assay provides a new tool for simultaneous quantitative catecholamine determination and was successfully applied in routine for the screening or follow up of pheochromocytoma, paraganglioma and neuroblastoma. LC-HRMS method offers significant advantages compared to LC-EC with good sensitivity, an unambiguous analyte determination and high sample throughput.

尿儿茶酚胺及其甲基化代谢物是嗜铬细胞瘤，副神经节瘤和神经母细胞瘤的生化指标。建立了一种基于固相萃取（SPE）和液相色谱分离与高分辨率质谱（LC-HRMS）的快速，精确的分析方法，并已验证该方法可测量尿中的儿茶酚胺（肾上腺素（E），去甲肾上腺素（NorE），多巴胺（D））和总甲基化代谢产物（去甲肾上腺素（NorMN），间肾上腺素（MN）和3-甲氧基酪胺（3-MT））。使用这种LC-HRMS方法测量的51个尿液样本的结果与带有电化学检测的液相色谱分析（LC-EC）进行了比较。尿液样品（200μL）掺有内标溶液，然后进行SPE纯化。对于总甲基化代谢物，在纯化SPE之前，尿液先被水解。在Acclaim混合模式WCX色谱柱上实现了8.5分钟的分离。对所有化合物均采用电喷雾正电离模式进行检测，并进行平行反应监测，并对E和NorE分别进行2至200 µg / L（10.9-1090; 11.8-1182 nmol / L）之间的线性回归（r2> 0.998）定量其他浓度则在10和1000 µg / L之间（D，M，NorMN和3-MT分别为65.2-6520; 50.7-5070; 54.5-5450; 59.8-5980 nmol / L）。总体不精确度和偏倚不超过15％。没有观察到明显的基质效应。除肾上腺素外，两种测定之间的相关性良好。通过LC-EC方法测量的肾上腺素浓度略高于通过LC-HRMS方法获得的值，但对临床决策没有影响。该LC-HRMS测定法提供了同时定量儿茶酚胺测定的新工具，并已成功应用于例行嗜铬细胞瘤，副神经节瘤和神经母细胞瘤的筛查或随访。与LC-EC相比，LC-HRMS方法具有显着的优势，具有灵敏度高，分析物确定明确，样品通量高的优点。