i am confused if sample should be prepared in mobile phase as used in HPLC Will the different solvent for sample and mobile phase being different affect peaks or any component of anlysis ?
Good question. That is not the best approach to us. Following good chromatography fundamentals, you should always try to dissolve the sample in the same mobile phase used. If a gradient is used, then use the gradient "initial conditions" composition. This insures the sample will be loaded to head of the column as a compact peak, with less injection artifact signals and improved peak shape. *Consider taking a basic HPLC intro class where these types of important questions are addressed.