The addition of propranolol induced the aggregation of gold nanoparticles, and increased Rayleigh light scattering (RLS) intensity greatly. The interaction between them was studied by RLS spectrum, UV-Vis spectrum and transmission electron microscopy (TEM). Based on these results, a novel method was proposed for propranolol assay. With the combination of solid phase microextraction (SPME), the proposed method was successfully applied to determine propranolol in urine.