High-performance liquid chromatography (HPLC; formerly referred to as high-pressure liquid chromatography) is a technique in analytical chemistry used to separate, identify, and quantify each component in a mixture. Advantages of use of ultra and nano High Performance Liquid Chromatography is : increased mass sensitivity owing to the reduced chromatographic dilution,the use of smaller particle sizes (<3 μm), lower stationary phase, solvent, and sample volume consumption, and the compatibility with flow rate requirements of a nanospray interface for on-line coupling to mass spectrometry (MS).