For the first time, a sensitive and selective method for determination of docetaxel (as an anti-cancer drug) at gold nanoparticle-multiwall carbon nanotubes/glassy carbon electrode (Au-MWCNTs/GCE) was suggested using cyclic voltammetry and differential pulse anodic stripping voltammetry (DPASV) method. After the construction of the electrochemical sensor and optimization of the effective parameters such as pH, accumulation time, and potential, the sensor was applied for the determination of docetaxel in the range of 0.3–3.3 μmol L−1. The results show that Au-MWCNTs significantly catalyzed the redox reaction of docetaxel during electrochemical detection. The limit of detection was estimated to be 90 nmol L−1 based on 3Sb/m. The performed studies showed that Au-MWCNTs/GCE has a good selectivity, sensitivity, and reproducibility. This sensor was used to determine the docetaxel in real samples (human urine and human serum), and the obtained data illustrate that fabricated electrochemical sensor is promising for use in routine analytical applications. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature.