HPLC is a technique used to separation, identification, quantitative and for qualitative analysis of each analyte in a sample depends on its interaction with the stationary phase and polarity. Pump flow a mobile phase by high pressure with the sample mixture through a column and allow for better separation, so it is also called as high-pressure liquid chromatography. Each and every analyte in the sample interacts slightly with the adsorbent material, the analytes that interact strongly with the stationary phase will go slowly through the column than analytes with low interaction this different cause the separation rate of the different analytes.