Simultaneous quantification of tracheloside and trachelogenin in rat plasma using liquid chromatography/tandem mass...

We developed and validated a quantitative method for simultaneously determining the concentrations of tracheloside and trachelogenin in rat plasma. Plasma samples were prepared by liquid-liquid extraction with ethyl acetate. Isocratic chromatographic separation was performed on a reversed-phase Diamonsil C(18) column (4.6×200 mm, 5 μm). The mobile phase consisted of methanol and 10mM aqueous ammonium formate (80:20, v/v). Analyte detection was achieved by positive electrospray ionization (ESI) tandem mass spectrometry. Calibration was performed by internal standardization with glipizide, and regression curves ranging from 0.625 to 625 ng/mL were constructed for both the analytes. The intra- and inter-day precision values were below 8%, and accuracy ranged from -5.33% to 2.53% in all quality control samples. In this study, the validated method was successfully applied to determine the pharmacokinetic profile of tracheloside and trachelogenin in rat plasma after oral and intravenous administration of trachelospermi total lignans.