β-Agonists are illegally consumed in various products such as food and animal and effect the nutrition distribution owing to change of body fat. In addition, they result in acute poisoning and several symptoms such as muscular tremors and nervousness. A new electrochemical approach based on two-dimensional hexagonal boron nitride (2D-hBN) nanosheets decorated functionalized multi-walled carbon nanotubes (f-MWCNTs) was presented for simultaneous determination of β-agonists such as phenylethanolamine A (PEA), clenbuterol (CLE), ractopamine (RAC) and salbutamol (SAL) in urine samples. X-ray diffraction (XRD) method, Raman spectroscopy, scanning electron microscope (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used characterizations of nanomaterials. After that, 2D-hBN/f-MWCNTs nanocomposite modified glassy carbon electrode (GCE) was prepared for simultaneous determination of β-agonists. 1.0 × 10-12-1.0 × 10-8 M and 1.0 × 10-13 M were founded as the linearity range and the detection limit (LOD) for PEA, CLE, RAC and SAL. Finally, the prepared electrochemical sensor was used for urine sample analysis in presence of ascorbic acid (AA) and uric acid (UA).