The aim of the study was to adapt the method for the determination of vitamins E, A, and β-carotene and to assay them quantitatively in plasma of municipal transport drivers. The study embraced 147 municipal transport male drivers, aged 23-58 years. The Waters 2695 integrated analytical system of high-pressure liquid chromatography (HPLC), equipped with a UV-VIS detector was used to determine the studied compounds. Our analysis of the quantitative data by age and seniority did not show significant differences in the concentrations of the analyzed compounds between the study groups, except for the concentration of β-carotene, which was significantly lower in drivers aged over 46 years with the longest employment (over 16 years) compared to the younger groups.