An LC–MS method for the determination of Darifenacin (DRN) in human plasma was developed and validated. Sample preparation involved the extraction with Diethylether: Dichloromethane:: 80:20, v/v) and chromatographic separation was performed on a CC 150 x 4.6 NUCLEOSIL 100-5 NH2 column with the mobile phase consisting of acetonitrile: milli-Q water: formic acid (90:10:0.1 v/v/v). The interface used with the API 3000 LCMS/ MS was a turbo ion spray in which positive ions were measured in MRM mode. The method was validated over the concentration range of 0.025- 10.384 ng/mL. The recovery was 90.94% -109.89% and the limit of quantitation (LOQ) was 0.025 ng/mL. The intra- and inter-day precision of the method at three concentrations was 0.84-2.69% and 2.01-7.47% while the intra- and inter-day accuracy was 94.79-108.00% and 102.61 -94.63%. Stability of compound was established in a series of stability studies. The method was successfully applied on bioequivalence study of extended release DRN tablet to obtain the pharmacokinetic parameters.