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Standard sunflower oil contains predominantly alpha-tocopherol (>95%). The discovery by Demurin (1993) that within the sunflower collection there are examples of spontaneous mutations when it comes to tocopherols has made it possible manipulate tocopherol types and contents in the sunflower oil. Used in the study were genetic materials obtained by crossing the standard line HA-26-B with isogenic line BK-66-TPH-2-OL-1. After inbreeding F1, F2, F3 and F4, the F4 population was selected and grown in the greenhouse of the Institute of Field and Vegetable Crops in Novi Sad, Serbia. During the flowering, each individual plant was isolated with a natron bag in order to enable self-fertilization. The qualitative and quantitative tocopherol contents were determined by the HPLC method developed at our laboratory, according to the offi cial ISO 9936 method, on the Sykam HPLC system coupled with the both, UV and fluorescent detectors, and supported by the Clarity Chromatography Data Station (CDS) software for data acquisition and processing. Separation of tocopherols in the oil was carried out by the use of normal-phase chromatography procedure with the binary eluent n-hexane : ethyl acetate in proportion 75 : 25, and flow of the eluent through the chromatographic column at 1 ml/min.