Direct infusion electrospray ionization-ion mobility-mass spectrometry for rapid metabolite marker discovery of medicinal Phellodendron Bark.

Ion-mobility mass spectrometry (IM-MS) currently serves as a powerful tool for the structural identification of numerous biological compounds and small molecules. In this work, rapid metabolomic analysis of closely-related herbal medicines by direct injection ion mobility-quadrupole time-of-flight mass spectrometry (DI-IM-QTOF MS) was established. Phellodendron chinense Bark (PC) and Phellodendron amurense Bark (PA) were studied as a case. Thirty-three batches of PC and twenty-two batches of PA have been directly injected in electrospray ionization-IM-QTOF MS in positive mode. Without chromatographic separation, each run was completed within 3 min. After data alignment and statistical analysis, a total of seven chemical markers were found (p-value < 0.05, VIP > 1.00). Among them, the ion m/z 342.17 and m/z 356.18 present a single peak in the drift spectrum, respectively, but their drift time has a certain deviation compared with the pure substance of known compounds. In addition, the MS/MS spectra also confirmed that the single peak includes two chemical isomers. To investigate the composition ratio of individual isomers, the calibration curves of relative drift time (rDT) based on the standard superposition method were established, which were found to fit the least square regression. The ion [M]m/z 342.17 was recognized consisting of magnoflorine (MAG) and phellodendrine (PHE), and their composition ratio in PA and PC samples was calculated. The results were compared with those obtained by the HPLC quantitative method, which produced equivalent quantification results. Our DI-IM-QTOF MS methodology provides an additional methodology for the relative quantification of unresolved isomers in drift tube IM-MS and offers DI-IM-QTOF MS based metabolomics.