The aim of this study is the using and performance controling of Hypersil ODS Column and Florescence dedeklor with HPLC. With the same phase and with different program (Vitamin A: Ex; 340nm-Em: 460, Vitamin E: Ex: 298-Em: 350). Vitamin A and E were detected and to this target Hypersil ODS reversed phase column was found useful in this work. With its filling material (5 µm, RCM 100 RP18. reversed Phase) column is very sensitive and has a speciall highseparator peculiarity. Vitamins, especially latsoluble vitamins: has more difficulties to extract and to determine. HPLC is used since seventies in the animal, food .medical and chemical industry and has more difficulties with its filling material and capacity of colons. Technical datum of column is very variable with flow liquid, and detected material. In this work we found timing of vitamins and their area and performans of column with flow rate of HPLC. With this result consantrations can be determined.