Tocopherols (Ts) and tocotrienols (T3s), collectively known as tocols, are natural compounds of plant origin. These compounds are biologically active fat-soluble antioxidants, which are physiologically very important in human and animal diets. Tocols (commonly known as vitamin E) possess very important health promotion properties like preservation of the membrane structure and integrity of cells, antioxidation, regulation of expression of genes involved in inflammation and immune system disorders, anticancer activity, regulation of blood cholesterol contents, regulation of blood clotting and increase of Ca bioaccumulation in tissues. Due to the positive correlation between dietary Ts and T3s and the human health and their influence on stability and shelf-life of antioxidants-fortified food products it is of utmost importance to be able to determine trace amounts of tocols in different dietary sources. Thus, the purpose of the studies was to optimize the very sensitive and versatile pre-column procedures that would allow to achieve the maximal concentrations of assayed Ts and T3s in various biological samples. Fortunately, the satisfactory sensitivity and separation of all forms of T and T3 was obtained using photodiode detection and columns filled with conventional C18-stationary phases. Moreover, the combination of esterification of tocols, C18-liquid chromatography and UV monitoring at 278 and 205 nm provide the suitable analytical tool for quantification of β- and γ-forms of tocols in biological samples. Our original C18-chromatographic methods are satisfactory precise, accurate, repeatable and offer low values of limit of detection (