A sensitive and selective LC-MS/MS method for the quantitative determination of segetalin A from the plasma of rats

A sensitive and selective LC-MS/MS method was developed and validated for the determination and pharmacokinetic investigation of segetalin A in rat plasma. Sample preparation was accomplished through a simple SPE procedure for the removal and preconcentration of the analyte and IS. Plasma samples were separated by HPLC on a Symmetry C18 column using a mobile phase consisting of methanol and 0.1% formic acid in water (70:30, v/v) with isocratic elution. The quantification was performed using multiple reaction monitoring with the transitions m/z 610.3 → 511.2 for segetalin A and m/z 779.4 → 751.4 for IS, respectively. The calibration curve was linear over the range of 8.0-4000 ng/mL with a limit of quantitation (LOQ) of 8.0 ng/mL. This method was applied in a pharmacokinetic study of segetalin A in rats. For intravenous (i.v.) administration, the plasma concentrations of segetalin A decreased quickly (t1/2z, 1.31 ± 0.341 h). For oral administration, the plasma concentrations of segetalin A increased to a peak value at 1.50 ± 0.577 h, followed by a gradual decrease to the LOQ in 12 h. The mean AUC values after i.v. and oral administration were 553 ± 105 and 1482 ± 110 ng h/mL, respectively.