Simultaneous determination of liensinine, isoliensinine and neferine in rat plasma by UPLC–MS/MS and application of the technique to pharmacokinetic studies

Ethnopharmacological relevanceThe in vivo effects of traditional herbal medicines are generally mediated by multiple bioactive components. The main constituents of Lotus Plumule are alkaloids such as liensinine, isoliensinine and neferine. In this study, a simple, sensitive, and robust analytical method based on ultra-performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) has been developed for the determination of the three alkaloids in rat plasma using carbamazepine as internal standard (IS).Materials and MethodsAfter precipitation of the proteins with acetonitrile, chromatography was performed on an Acquity UPLC BEH C18 column (2.1 mm×50 mm, 1.7 μm particle size) using a gradient elution with 0.1% formic acid in water and acetonitrile. Mass spectrometry involved positive electrospray ionization and multiple reaction monitoring (MRM) of the transitions at m/z 611.7→206.2 for liensinine, 611.3→192.2 for isoliensinine, 625.2→206.1 for neferine and m/z 237.1→194.2 for IS.ResultsThe method was linear over the concentration range 5–1000 ng/mL with a lower limit of quantifof 5 ng/mL for each alkaloid. Inter- and intra-day precision (RSD%) were all within 11.4% and the accuracy (RE%) were equal or lower than 10.4%. Recoveries were more than 75.3% and matrix effects were not significant. Stability studies showed that the three alkaloids were stable under a variety of storage conditions.ConclusionThe method was successfully applied to a pharmacokinetic study involving intravenous administration of liensinine, isoliensinine and neferine to rats.

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