High performance liquid chromatography (HPLC) is an important qualitative and quantitative technique, generally used for the estimation of pharmaceutical and biological samples. HPLC is the most often used separation technology for detecting, separating, and quantifying the drug. HPLC technique development and validation serve critical roles in novel drug discovery, development, and manufacturing, as well as a variety of other human and animal investigations. HPLC instrumentation includes a Solvent reservoir, pump, injector, column, detector, and integrator or acquisition and display system. The heart of the system is the column where separation occurs. The information that can be obtained using HPLC includes identification, quantification, and resolution of a compound.