Vitamins are a well-known a group of compounds that are essential for human health.And either their lack or excess can cause health problems.Vitamins A (retinol) and E (tocopherol) are usually added as their acetate forms.Vitamin D is added as either the D3 (cholecalciferol) or D2 (ergocalciferol) and the amount of vitamin D is much lower than vitamin A and E.Fat-soluble vitamins can be analyzed by normal phase or reversed phase HPLC.Reversed phase HPLC is a technique well suited for these because of easier for use and lower cost.Reversed phase HPLC is used by China national food safety standard for analysis vitamin A, D and E (see Determination of vitamin A, D, E in foods for infants and young children, milk and milk products, 2010).