A rapid, simple, selective and sensitive LC-MS/MS method was developed for the determination of Teicoplanin in human plasma using Imipramine as internal standard (IS). The method was developed with turbid ion spray (TIS) in the positive ion and multiple reaction monitoring (MRM) mode. The assay procedure involves a simple liquid – liquid extraction of Teicoplanin and Imipramine from human plasma by using methyl-tert-butyl ether. The mobile phase was acetonitrile: 10mM ammonium formate, pH 4.5 in the ratio of 90:10%v/v. Chromatographic separation was achieved on Gemini C18 (50×4.6mm,5µ) column with a flow rate of 1.0ml/min. The MRM transitions monitored for Teicoplanin and Imipramine were 324.70/108.90 and 280.80/86.00 respectively. The developed method was validated as per FDA guidelines. Linearity was observed from 306.022- 199205.354pg/ml with correlation coefficient of 0.9969. The percent recovery for the drug and IS was found to be 78.31 and 64.96% respectively. Stability studies like freeze thaw, bench top, short term and long term were performed and the results were found to be within the acceptance limits according to FDA guidelines.