Simultaneous Determination of Kaempferide, Kaempferol and Isorhamnetin in Rat Plasma by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry and its Application to a Pharmacokinetic Study

Abstract

A selective, rapid and sensitive method using ultra-high performance liquid chromatography was developed and used for the simultaneous determination of kaempferide, kaempferol, and isorhamnetin in rat plasma after oral administration of Sedum sarmentosum Bunge extract. A 6430 triple-quadrupole tandem mass spectrometer in multiple reaction monitoring mode, with a negative electrospray ionization source, was used for the detection. A Waters Symmetry C18 column with isocratic elution (methanol:5 mM ammonium acetate at 75:25, v/v) was used for separation at a flow rate of 0.3 mL min -1 . The method was linear for all analytes over the investigated concentration range (r > 0.9900). The intra-and inter-day precisions ranged between 3.88 and 7.74% and the accuracies ranged between 88.63 and 98.74%. The mean recoveries and matrix effect were higher than 85.66 and 92.36%, respectively. All the three compounds tested were stable during storage and analyses. The method was successfully applied to a pharmacokinetic study.