The fluorescence emission from the lithium/1,4-dihydroxyanthraquinone system shows a great enhancement in the presence of certain water-miscible solvents. This is justified from the donicity (nucleophilic properties) of the solvent that facilitates the solvation of the lithium cation in solution and the stabilization of an nondissociated ion-pair between the solvated lithium cation and the 1,4-dihydroxyanthraquinonate anion. A very sensitive analytical method was proposed for the spectrofluorimetric determination of lithium based on its reaction with 1,4-dihydroxyanthraquinone (quinizarin) in a dimethylsulfoxide medium (90%) and in presence of sodium hydroxide. The fluorescence is measured at an excitation wavelength of 602 nm and an emission wavelength of 670 nm and it is stable at 25°C at least 6 h. The calibration curve is linear over the concentration ranges of 2–40 μg/l of lithium in an aqueous matrix and 3–50 μg/l in a serum matrix; the RSD's in the determination of 20 μg/l of Li+were 2.6% and 3.2%, respectively. The proposed procedure was satisfactorily applied to the determination of lithium in drugs, dietetic products and human serum.