Establishment of quantitative methodology for sophoridine analysis and determination of its pharmacokinetics and bioavailability in rat

Objective The aim of this study is to develop a rapid and sensitive UPLC-MS/MS approach to determine the sophoridine (SOP) level in rat plasma and the pharmacokinetics of the substance. Significance Sophoridine is used as an anti-inflammatory, anti-virus, anti-microbial, and anti-tumor alkaloid. It is essential to explore specific detection methods for the quantitative analysis of SOP in the blood circulation. Methods The rat plasma samples were prepared by one-step protein precipitation with acetonitrile. Subsequently, the samples were separated by chromatography using a UPLC BEH C18 reversed-phase with an initial mobile phase of methanol and 0.1% formic acid aqueous solution. The gradient elution was performed at a fixed flow rate of 0.4 mL/min, and multiple reaction monitoring (MRM) mode with an electrospray positive ionization source was employed to detect the transitions of m/z 249.1 -> 84.2 for SOP and m/z 264.3 -> 69.8 for dendrobine (IS). The entire process required 3.5 min for each sample. Results A linear correlation was established over the range of 2-2000 ng/mL (r (2)>= 0.9954) for SOP in rat plasma with a lower limit of quantification (LLOQ) at 2 ng/mL. The range of accuracy was tested between 94.90% and 100.80%, and the relative standard deviations (RSDs) toward both intra- and inter-day precision were <10%. Thus, this method was successfully applied to a pharmacokinetic study, and the subsequent results demonstrated a low absolute bioavailability of 2.32%. Conclusion The present study established a reliable method that quantified the SOP concentration in rat plasma after administering a dose of 2 mg/kg intravenously or 20 mg/kg orally.

Automated summary

This study developed a rapid and sensitive UPLC-MS/MS approach to determine SOP levels in rat plasma and the pharmacokinetics of the substance. The method showed a linear correlation over a range of 2-2000 ng/mL with good accuracy and precision, and it was successfully applied to a pharmacokinetic study revealing a low absolute bioavailability of 2.32%.