Waters ACQUITY UPC2 System with Trefoil AMY1 and CEL1 Columns provide a simple and fast solution for differentiation between the natural vitamin E and the synthetic vitamin E. The stereoisomers of α-tocopherol and its acetate can be separated into 2 or more peaks. Based on the chromatographic profile, natural vitamin E can be easily differentiated from synthetic vitamin E. The sample preparation for these analyses is simple. There is no derivatization required. The chromatographic analysis run time is 35 min for the α-tocopherols, and 15 min for the α-tocopheryl acetate, respectively. For the first time, the differentiation between natural and synthetic vitamin E can be easily conducted. These analytical methods are suitable for the routine analysis of vitamin E in foods, dietary supplements, and other related products.