Fluorescence characteristics of complex system of rare earth terbium (Tb3+), norfloxacin, 1,10-phenanthroline(1,10-phen) and sur- factant 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-Tb3+ (NFLX-Tb3+) 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-Tb3+ 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 CNFLX+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 sim- ple, rapid, sensitive, practical and suitable for direct analysis of pharmaceutical preparation. It was successfully applied to determine the cap- sules, human serum and urine in real pharmaceutical samples.