A novel preconcentration/separation method for simultaneous sorption-spectrophotometric determination of anionic food dyes Sunset Yellow and Tartrazine is proposed. The method is based on preconcentration of the dyes using solid phase extraction on a cartridge filled with silica chemically modified with C16 groups from aqueous solution at pH 1 followed by elution with water/acetonitrile mixture containing 2 mmol·L−1 KH2PO4 adjusted to pH 3 with a step gradient of acetonitrile content. This elution allows quantitative separation of the dyes which makes their individual spectrophotometric determination possible. The detection limits for Tartrazine and Sunset Yellow are 0.15 and 0.11 μg·mL−1 and the linearity range is 2–20 μg·mL−1. The method is applied for analysis of beverages. The recovery of dyes is higher than 97% at the relative standard deviation not exceeding 10%.