In the present study, a rapid, practical, selective and sensitive method was proposed for simultaneous determination of dopamine (DA) and uric acid (UA). Kinetic profiles of analytes were recorded and transformed by different mother wavelets. The contribution of ascorbic acid was neglected using mean centering of data before transformation. The concentrations range were 1.0–60.0 μmol L−1 and 7–40.0 μmol L−1, with detection limit of 0.06 and 0.30 μmol L−1 for uric acid and dopamine, respectively. The results of sym8 and db4 mother wavelets showed that proposed method creates a most selective and sensitive determination without using initial separation steps. The obtained results by CWT-sym8 and CWT-db4 were compared with partial least squares (PLS) results. The simultaneous quantitation of DA and UA with the proposed method was successfully applied in different urine and serum samples.