The large-scale controllable, ordered two-dimensional arrays of gold nanostructure with hot-spot were prepared together with chemical molecules were modified on the surface to concentrate SudanⅠwithin the zone of the SERS effect, which lead to analytical detection of SudanⅠin high resolution. The vapor of gold was deposed on anodic aluminum oxide(AAO)template by ～200 nm thickness to replicate its nanochannels, and the negative structure i.e. large-scale ordered gold nano-hemisphere array, was obtained after the removal of the template of AAO by NaOH solution. Au nano-hemisphere array was modified by 1-Dodecanethiol which can be self-assembled monolayer on the surface and concentrate SudanⅠwithin the zone of the SERS detection, which can facilitate the measurement of SudanⅠ. Due to the order and regularity of Au nano-hemisphere array, the signal of SudanⅠin the range of laser illumination is stable and uniform, and the quantitative analysis of SudanⅠwas realized. The SERS intensity of SudanⅠis logistic proportional to the concentration in the range of 10-7 to 5×10-4mol·L-1. The corresponding correlation coefficient of the liner equation is 0.99, the recoveries of SudanⅠare between 77%～117%. The limit of detection for SudanⅠis 4×10-8mol·L-1, comparable to that of HPLC of Chinese national standard method.