Food dyes are one of the most widely used additives in commercial food products such as beverages, candies, pharmaceuticals, and cosmetics. Despite the laws and regulations which aim to limit the synthetic colourant usage due to their pathogenic effects, they are widely used. Therefore, analyzing these dyes become very important for human health. Tartrazine is one of the commonly used synthetic food dyes and so the development of a new, simple, and accurate separation, preconcentration, and spectrophotometric determination of tartrazine (E 102) was the aim of this work. The method was based on solid-phase extraction by using Diaion SP-207 resin and determination with UV Spectrophotometer. Analytical parameters affecting the recoveries of tartrazine were investigated to find the optimal conditions. The retained tartrazine was removed from the column by 10 mL of 0.01 M HCl in acetone. The recovery values were found as >95%. By this new method, tartrazine contents of various food and medicine samples were determined successfully. TThe recoveries of understudy analytes were found in the range of 93–98%, while the detection limits were calculated in the range of 0.3–2.0 μg L−1.he detection limit was found as 3.52 μg/L. Calibration standards used in the experiments were in the range of 10-7 -1x10-5 mol L-1 with a correlation coefficient of 0.999.