Development and validation of an UPLC-MS/MS method for determination of jujuboside B in rat plasma and its application in pharmacokinetic and bioavailability studies

Abstract

Jujuboside B (JuB) is a main bioactive saponin constituent of Ziziphi Spinosae Semen. The compound is used clinically as an anti-insomnia and anti-anxiety medicine. In this study, a sensitive, simple, and rapid ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) was developed and validated to quantify JuB in rat plasma. A simple protein-precipitation method was used to extract JuB from rat plasma samples. Jujuboside A was used as an internal standard (IS). Chromatographic separation was performed using an Acquity HSS T3 column. The mobile phase consisted of acetonitrile and 0.1% formic acid in water with a flow rate of 0.3 mL min⁻¹. Identification and quantification were performed through electrospray ionization in a negative mode with multiple reaction monitoring of JuB and IS transitions of m/z 1043.3 → 911.5 and m/z 1205.6 → 1073.4, respectively. The calibration curve was linear in the range of 0.1 ng mL⁻¹ to 1000 ng mL⁻¹ (R² = 0.990) with a limit of detection of 0.03 ng mL⁻¹. The extraction recoveries of JuB were 90.3% to 95.7% and the precisions of intra-and inter-day were less than 11.5%. The matrix effect of JuB at three different concentrations ranged from 93.5% to 95.9% with a standard deviation of <5%. The inter- and intra-day assay accuracies were 86.7% to 94.3% and 93.3% to 95.7%, respectively. The pharmacokinetic processes of JuB fit in the one-compartment model of p.o. administration and two-compartment model of i.v. administration. The absolute oral bioavailability of JuB in rats was only 3.6%. Indeed, the proposed method was successfully applied to analyze the pharmacokinetics and bioavailability of JuB in rats after JuB was administered p.o. and i.v.