Description:
A simple and sensitive liquid chromatographic method along with tandem mass detection has beendeveloped for the determination of levodopa and carbidopa in mice plasma. Owing the hydrophilicnature of both the analytes, a hydrophilic interaction liquid chromatography (HILIC) setup was usedfor their separation by a Merck (Germany) HILIC column (4.6 × 250 mm, 5 μm; 200 Å). The mobilephase composed equal proportion of water and acetonitrile both containing 0.1% formic acid at a flowrate of 1.4 mL/min to achieve rapid separation of the compounds. The column was coupled with atriple quadrupole mass spectrometer equipped with an electrospray ionization (ESI) source usingmulti reaction monitoring (MRM) analysis. Since, recovery of both analytes in mice plasma posedsignificant challenge, a customized extraction procedure based on protein precipitation was adoptedwith best recovery. The optimized HILIC-MS/MS condition led to yield lower limit of quantification(LLOQ) of 9.9 ng/mL and 2.47 ng/mL for levodopa and carbidopa, respectively. The method wasvalidated with suitable determination of correlation coefficient (R2: 0.997), precision (1.6-17.2%),accuracy (84.7-120%). Successful application of this validated method was accomplished for analytesin biological samples.