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Catecholamines are produced by the sympathetic nervous system or by the medullary adrenal glands and play a key role both as hormones and as neurotransmitters. An increase in their production can lead to hypertension, due to neuroendocrine tumors like pheochromocytoma, paraganglioma and neuroblastoma. Among tumors of neuroendocrine derivation, there are also carcinoids that can cause carcinoid syndrome, associated with an excess of serotonin release. Proper biochemical analysis of patients suffering from these tumors is crucial to confirm the presence of excess production of catecholamines or serotonin and their metabolites. Serious and life-threatening cardiovascular complications related to cathecolamines and their metabolites pose a challenge to the laboratory for a timely and accurate diagnosis of neuroendocrine tumors. The application of innovative technologies to determine these analytes, such as LC-MS/MS, potentially associated with a metabolomic approach, may provide an efficient tool to help physicians recognizing and appropriately treating these still difficult-to-find clinical entities. As a complement to this paper, we present a personal experience about the analysis of biogenic amines and their metabolites performed on both 24-hour and spot urines. The LC-MS/MS method is used with three different preparation phases: total metanephrines (free and conjugate), catecholamines, free methanephrine and serotonin and vanillylmandelic acid (VMA), homovanillic acid (HVA), 5-hydroxyindoleacetic acid (5-HIAA). The organization of the chromatography and mass spectrometry division, based on expert technicians, includes centralized acceptance of samples, automation of the preparation phase and middleware-based validation of the results, offering to clinicians a valuable help in this challenging diagnostic. [ABSTRACT FROM AUTHOR]