Background: Vildagliptin, a dipeptidyl peptidase-4 inhibitor, is one of the potent oral antidiabetic agent. Objective: The objective of the present work was to establish a rapid, sensitive and validated capillary electrophoresis Quadrupole Time-of-flight Mass Spectrometry method from rat plasma. Methodology: Vildagliptin was estimated in rat plasma after precipitation of plasma proteins by acetonitrile, using sitagliptin as internal standard. For separation of vildagliptin from plasma components, fused silica capillary with background electrolyte consisting of 0.25 mM ammonium formate buffer, with SL composition of 50:50 methanol and water consisting of 0.25% of formic acid, pumped at a flow rate of 0.2 ml/min was used. Electron spray ionization with positive ion multiple reaction mode was applied for detection of analytes. Results: Newly developed method showed good calibration curve in the concentration range of 1 -500 ng/ml with excellent correlation coefficient (r