This paper presented a fast screening method for determining Sudan dye adulteration in grape juice, based on the combination of High Performance Thin-Layer Chromatography (HPTLC) and Surface Enhanced Raman Spectroscopy (SERS). Chromatography separation was achieved on silica gel plates using petroleum/acetone/acetic acid (9/1/0.1) as the mobile phase. After that, key factors including silver nano particle concentrations, electrolyte types/concentrations and incident laser wavelength was investigated with regarding to their effects on the quality of SERS results. Under optimized conditions, the method sensitivity reached to 0.01～0.05 mg/kg, while remarkable linearity (R²＞0.996) was obtained within the range of 1～20 mg/kg. Apart from that, the established method showed satisfactory accuracy (standard-spiking recovery rates 72%～104%) and precision (RSD 4.5%～9.6%) in the validation with the grape juice sample.