An efficient and accurate Reverse Phase – High Performance Liquid Chromatographic method was developed and validated for the separation and determination of synthetic food colorant Tartrazine(E102)in certain food stuff and in drugs. This method was successfully applied for the estimation of tartrazine dye in certain food stuff (cream biscuits, Tang, gems, and candies) and in drugs (Aceclofenac and Metoprolol tablets - in which tartrazine was present as a coating agent). Interaction study was carried out to find the effect of tartrazine on the protein binding of Drugs in Bovine serum albumin. The objective of this research was to estimate the tartrazine dye in various food stuff and formulation, and to determine the effect of dye on drug.Tartrazine extracted and quantified in various food stuff. The amount of E102 present in Aceclofenac and metoprolol tablet was also estimated. A study on protein binding of drugs by UV spectroscopy and RP-HPLC were conducted.The effect of tartrazine on protein binding of Aceclofenac and Metoprolol in BSA and were carried out and determined that Protein binding of the drugs was altered by the effect of dye.