Bhupinder Singh, Naveen Dubey, Peeyush Jain, Sandeep Sharma, Ashish Dwivedi, Rama S.lokhandae
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.
