The rapid profiling and simultaneous determination of 12 major alkaloids in Nauclea officinalis by UPLC-Q-TOF-MS and HPLC-ESI-MS/MS

Abstract

For the first time, we determined the phytochemical profile of alkaloids from N. officinalis using ultraperformance liquid chromatography-quadrupole-time-of-flight-tandem mass spectrometry (UPLC-Q-TOF-MS/MS). A rapid, specific, and sensitive high-performance liquid chromatography-tandem mass spectrometry method was subsequently developed and fully validated for the simultaneous determination of 12 major alkaloid constituents in N. officinalis: pumiloside; naucleoxoside A; naucleoxoside B; nauclefine; angustidine; angustoline; (3S,19S)-3,14-dihydroangustoline ([α]²⁰_D: (−)191°); (3S,19R)-3,14-dihydroangustoline ([α]²⁰_D: (−)294.7°); strictosamide; angustine; vincosamide; and 3,14-dihydroangustine. The analytes were detected with an electrospray ionization (ESI) source and multiple reaction monitoring (MRM) using positive scanning mode. Three pairs of epimers (naucleoxoside A and naucleoxoside B; (3S,19S)-3,14-dihydroangustoline ([α]²⁰_D: (−)191°) and (3S,19R)-3,14-dihydroangustoline ([α]²⁰_D: (−)294.7°); and strictosamide and vincosamide) were successfully separated using an ACQUITY UPLC® BEH C₁₈ column (2.1 mm × 100 mm, 1.7 μm) at 30 °C, with a gradient mobile phase consisting of 0.1% formic acid aqueous solution (A) and acetonitrile (B), a flow rate of 0.2 mL min⁻¹, and a total run time of 30 min. All calibration curves exhibited excellent linear regression (r > 0.999) within the test range. The precision, repeatability, and stability of the method toward the 12 alkaloid compounds were less than 2.0% in terms of the relative standard deviation (RSD) values. The mean recoveries for all compounds were between 98.7% and 101.1%, with RSD values ranging from 0.55% to 1.7% for N. officinalis samples.