Determination of Isotoosendanin in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry: Application to Pharmacokinetics Study

A rapid, sensitive and accurate liquid chromatography tandem mass spectrometry (LC-MS-MS) method was developed and validated for the quantification of isotoosendanin, an important bioactive component isolated from Meliae cortex, A Capcell PAK C18 column (100 x 4.6 mm) was used for the chromatographic elution using methanol-10 mM ammonium acetate-formic acid (80:20:0.1, v/v/v) as mobile phase at the flow rate of 0.6 mL/min. MS-MS analysis was performed on a triple quadrupole mass spectrometer equipped with an atmospheric pressure chemical ionization source in positive ion mode. Extraction of isotoosendanin and genistein (internal standard, IS) from rat plasma was determined by precipitating protein treatment. Quantification was performed by MS in the multiple reaction monitoring mode with positive ionization at m/z 557 -> 437 for the analyte and m/z 271 -> 215 for IS, respectively. Linear isotoosendanin calibration curves were obtained between 2.0-2,000 ng/mL with a correlation coefficient greater than 0.99. Acceptable precision and accuracy were acquired for concentrations over the standard curve range. Satisfactory results were achieved for sensitivity, specificity, recovery, freeze/thaw and stability. This analytical method was successfully applied to determine the pharmacokinetic parameters of isotoosendanin after an oral administration of 200 mg/kg to rats.