Abstract: A method of resonance Rayleigh scattering (RRS) was proposed for the determination of trace navelbine. In Britton-Robinson buffer at pH of 2.2, navelbine reacted with Eosin Y to form 1∶1 ionic association complexes. As a result, the intensity of resonance Rayleigh scattering was enhanced greatly and the new scattering spectra displayed the maximum RRS wavelength at 312 nm. The linear relationship was found between the increases of the scattering intensity (ΔIRRS) and the mass concentration of NVB in the range of 0.06-3.0 mg·L-1 with detection limit (3σ) of 1.7 μg·L-1. The method was applied to determine NVB in injection and urine samples, giving recovery rates measured by standard addition method in the range of 99.8%-101% and RSD (n=5) in the range of 1.1%-2.2%.