In this work and for the first time, it was developed a new methodology for detection of glibenclamide in spiked liquid samples (teas) by fluorometry (位ex = 300 nm; 位em = 404 nm). The novel methodology was also implemented in a miniaturized and portable automatic flow system based in the concept of multipumping with an in-line pre-separation unit. The separation of the drug from the liquid samples was achieved through adsorption of the drug into activated charcoal packed within a mini column followed by elution with a solution composed by ethanol, hydrochloric acid and the surfactant CTAB (70%, 1.0 mol L鈭?, 0.01 mol L鈭?, respectively). The results allowed to obtain a linear working range for glibenclamide concentrations of up to 50 mg L鈭? (r = 0.9999) and the detection limit was about 0.81 mg L鈭? of glibenclamide.