A Rapid and Sensitive Supercritical Fluid Chromatography–Tandem Mass Spectrometry Method for the Analysis of Celecoxib in Beagle Plasma: Application to Pharmacokinetic Studies

ABSTRACT To investigate the effect of formulation factors on the in vivo behavior of celecoxib (CXB) through pharmacokinetic tests, we developed and validated a supercritical fluid chromatography–tandem mass spectrometry (SFC–MS/MS) method for the analysis of CXB in beagle dog plasma. This method uses liquid–liquid extraction (LLE) to extract analytes from plasma, which is both economical and less susceptible to matrix effects. In order to increase sensitivity, improve peak shape, and accelerate elution, several key elements including stationary phase, mobile phase composition, column temperature, and back pressure, were investigated. The final analyte elution process on an ACQUITY UPC2 BEH column (100 × 3 mm, 1.7 µm) took only 2 min under optimal conditions. The selectivity, linearity ( r = 0.9980), precision (relative standard deviation: 3.8%–15%), accuracy (relative error: −11% to −0.7%), extraction recovery (86.0%–91.3%), matrix effect (91.2%–107.4%), and stability under different conditions of the method were all in accordance with the requirements. The method is stable, sensitive, green, and high‐throughput, and can be used for the determination of CXB in biological samples and the evaluation of its pharmacokinetic behavior.