The successful conclusion of human genome project partly owes its success to the advent of soft ionization techniques, namely electrospray ionization (ESI) and laser desorption ionization (MALDI), in mass spectrometry (MS). The recent advancements in MS technology in development of high resolution MS systems and coupling of separation units (Capillary Electrophoresis and Liquid Chromatography) with the soft ionization methods have enabled the use of mass spectrometry in the analysis biological and forensic systems. These applications include the drug discovery, biomarker identification for cancer, tuberculosis, proteomics and genomics. The research in mass spectrometry can be classified into two categories:'¢Development of novel instrumentation to improve the drawbacks (sensitivity, limit of detection, quantitative MS, etc) in the current methods '¢Biomolecular analysis to decipher the inborn errors in the metabolic pathway (Single Nucleotide Polymorphism, Post Translation Modifications, etc)The work that is presented in this dissertation falls into the first category. In the forthcoming chapters, we will focus on two aspects of novel microfluidic phenomenon of high frequency alternating current (AC) electrospray. The first portion of this thesis emphasizes on the comprehensive fundamental study (experimental/theoretical) for complete understanding of the mechanism that leads to the formation of AC electrospray. The second portion establishes the AC electrospray as a potential soft ionization method for mass spectrometry, with the added benefits of higher sensitivity, reduced ionization suppression and the frequency modulation of pH in the biomolecular analysis, which is direct contrast to the conventional DC electrospray.