A simple and highly sensitive method is proposed for the fluorimetric determination of nilvadipine in dosage forms and in biological fluids. The method involves the reduction of the nitro group of nilvadipine with zinc/HCl into a highly fluorescent product. The experimental parameters were carefully studied and incorporated into the procedure. The produced fluorophore exhibits its highest fluorescence intensity at λ_em of 458 nm after excitation at λ_ex of 356 nm. The relative fluorescence intensity-calibration plot was found to be linear over the range of 0.04-1.4 µg/ml with detection limit (S/N=2) of 6 ng/ml (1.56×10^-8 M) and correlation coefficient (n = 9) equal to 0.9999. The proposed method was applied to the determination of nilvadipine in its dosage forms. The results obtained were in good agreement with those obtained using a reference method. The high sensitivity of the proposed method allowed its further application for the in vitro determination of nilvadipine in spiked human plasma and urine. The percentage recoveries (n = 4) were 96.75 ± 0.60 and 99.09 ± 0.87, respectively.