LC-MS/MS determination of nevadensin in rat plasma and its application in pharmacokinetic studies.

A simple, sensitive and specific liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) method was developed and validated to quantitate nevadensin in rat plasma using genistein as the internal standard (IS). The assay was based on protein precipitation treatment with acetonitrile and liquid chromatography performed on a reverse-phase Zorbax SB-C(18) column (50 mm × 2.1 mm, 1.8 μm). Elution was achieved with a mobile phase consisted of aqueous solution containing 0.1% formic acid (solvent A) and methanol solution containing 0.1% formic acid (solvent B) at a flow rate of 300 μL/min (30:70, v/v). Quantification was through tandem-mass spectrometry with positive electrospray ionization (ESI) and multiple reaction monitoring (MRM) at m/z 345.1→315.1 and 271.1→215.1 for nevadensin and IS, respectively. Calibration curves were linear over the nevadensin rat plasma concentration range of 0.1-300 ng/mL. The lower limit of quantification (LLOQ) was 0.100 ng/mL and the inter- and intra-day accuracy and precision ranged between -1.67% and 7.60%. The average recovery of nevadensin was 95.6-99.1%. The validated method was successfully applied to the pharmacokinetic evaluation of nevadensin using the rat as an animal model following an oral administration of 50mg/kg nevadensin.