A new voltammetric method is proposed for high sensitive thiethylperazine (THP) determination, using a glassy carbon electrode modified with semi-graphitized carbon nanofibers/MnO nanocomposite (eCNF/MnO/GC). To the best of our knowledge, this is the first electrochemical assay of THP determination, and the first use of the eCNF/MnO as the electrode modifier. The proposed method using eCNF/MnO/GC is characterized by high repeatability and sensitivity of measurements, with the linearity of THP in the range from 0.05 to 2.2 µmol L−1. The lowest detection limit achieved on the eCNF/MnO/GC electrode for 30 s of preconcentration was 6.3 nmol L−1 THP in 0.05 mol L−1 acetate buffer of pH 5.6. The proposed method was successfully applied to highly sensitive THP determination in complex matrices, such as tablets and plasma with good recovery (98–103%). The RSD value obtained for THP measurement at a concentration of 0.1 µmol L−1 was 1.3%. Amperometric measurements of THP under the flow injection conditions were also performed to indicate the possibility of its fast and accurate determination (103% and 95% for unmodified and modified electrode, respectively), with the duration of single analysis of approx. 30 s.Graphical