Original Articles
Raghunadha Reddy. S1*, I. Sara
Abstract
A sensitive and selective method is described for the quantification of Ursodeoxy cholic acid in human plasma using Diclofenac as internal standard. The Bioanalytical approach consists of Precipitation with Acetonitrile, chromatographic separation of 10μL injected sample with Analytical Column (Thermo Bio basic C4, 5 μm, 150×4.6 mm) using 40:40:20 isocratic solvent mixture as mobile phase followed by quantification with mass detector in selective reaction monitoring mode using electro spray ionization mode (ESI) as an interface. The method was fully validated over a concentration range of 0.1 to 3.05 μg/ml using 0.1 ml of human plasma per assay. Stability assessment was also included. The run time for assay is 5 min. Robotic liquid handling systems are employed to all liquid transfer steps including the sample preparation procedure as well as to the addition/removal of the organic solvent. The current method includes a simple and rapid sample preparation as a result of robotic systems utilization that enabled parallel processing as well as shorter analysis run time compared to previously published methods. The method was applied successfully to the analysis of plasma samples obtained for pharmacokinetic, bioavailability or bioequivalence studies after therapeutic doses of Ursodeoxy cholic acid