Fluorescence characteristics of complex system of rare earth terbium (Tb 3+ ), norfloxacin, 1,10-phenanthroline(1,10-phen) and surfactant sodium dodecyl benzene sulfonate (SDBS) had been studied by using fluorescence spectroscopy. In a buffer solution (pH=6.3), the fluorescence intensity of the norfloxacin-Tb 3+ (NFLX-Tb 3+ ) system had been remarkably enhanced by SDBS and 1,10-Phen. When excited at 335 nm, a good linear relationship between fluorescence intensity of NFLX-Tb 3+ system and NFLX concentration was obtained in the range of 8×10 -8 -4×10 -6 mol/L, and the linear equation was F=2×10 8 C NFLX +22.7 with correlation coefficient of 0.9959. In addition, this method was compared with some previous literatures, the proposed method had relatively higher sensitivity and lower detect limit. The method was simple, rapid, sensitive, practical and suitable for direct analysis of pharmaceutical preparation. It was successfully applied to determine the capsules, human serum and urine in real pharmaceutical samples.