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A high performance LC-MS/MS method for determination of Metoprolol in human plasma was developed and successfully validated. Metoprolol is a β1-selective aryloxy propanolamine used in the treatment of cardiovascular disorders such as hypertension, arrhythmia and heart failure. Metoprolol (analyte) and the Metoprolol d7 (Internal Standard) were extracted from K3EDTA based Human Plasma samples by Liquid-liquid extraction method. Chromatographic separation was achieved on Gemini 5μm C18 110Å (150x4.6mm) column using Methanol: 10mM Ammonium Formate containing 0.1% Formic Acid Solution (95:5 v/v) as a mobile Phase at a flow rate of 0.800 mL/min with 80% flow splitting. The retention time of Metoprolol and Metoprolol d7 was found to be 1.49 minute and 1.49 minute respectively. The standard curve was linear (R2 > 0.99) over the concentration range of 0.68 ng/mL to 399.22 ng/mL. All the bioanalytical validation parameters were determined as per US FDA guidelines. The developed bioanalytical method was sensitive with a limit of detection below 1 ng/mL, with good linearity (r2 >0.99). The peaks obtained for the Metoprolol and Metoprolol d7 were symmetrical in nature and well resolved from each other without any interferences from human plasma. All the validation data, such as accuracy, precision, and percent recovery, lipemic effect, haemolysis effect, matrix factor, etc. were within the required limits as stated by guidelines. The method can be used for pharmacokinetic as well as Bioavailability, and bioequivalence studies. [ABSTRACT FROM AUTHOR]