An optical fibre (OF) analyzer for measuring catecholamines (dopamine, norepinephrine and epinephrine) in biological samples with induced fluorescence was developed. The analytical set-up included a chromatographic column for catecholamine separation and a fluorescence-based OF detection (FOF-analyzer). The detection limit of the FOF-analyser was found to be less than 0.9 pg mL -1. The proposed methodology showed an adequate analytical performance for the determination of the catecholamines in actual samples of human urine. The analytical performance of the FOF-analyzer for catecholamine determination was investigated against the high performance liquid chromatography-electrochemical detection (HPLC-ED) method. The FOF-analyzer showed lower detection limits and larger linear ranges for determination of dopamine (DA), norepinephrine (NE) and epinephrine (E) in comparison with HPLC-ED and other methodologies such as HPLC-fluorescence. These advantages combined with the compact design, small-scale instrumentation, and effective cost of analysis make this system an interesting alternative to the existing methodologies for the determination of catecholamines in clinical samples.