Rapid discrimination and quantification of isomeric flavonoid-O-diglycosides in Citrus paradisi cv. changshanhuyou by online extraction–quadrupole time-of flight tandem mass spectrometry

Abstract

Rapid differentiation, characterization and quantification of isomers from complex mixtures by direct mass spectrometry (MS) remained an analytical challenge due to their similar or identical MS/MS spectra and matrix interferences. Here, we reported a novel online extraction–quadrupole time-of-flight tandem mass spectrometry (OLE–QTOF-MS/MS) system to rapid, efficient and sensitive analysis of interglycosidic linkage isomers (hesperidin and neohesperidin) in Citrus paradisi cv. Changshanhuyou (Changshanhuyou). OLE system packed with solid Changshanhuyou (0.02 mg) could be firstly used to online remove interferences with large polarities, and then online extract and enrich hesperidin and neohesperidin, which shows great potential to diminish the analysis time of sample pretreatment, as well as to reduce matrix effects and instrument consumption. Detailed fragmentation analysis found that, under positive ion mode, relative abundance of specific fragment ions m/z 449 to m/z 303 showed linear correlation to the mass content of hesperidin (0% to 100%) with good correlation coefficient (R2 = 0.9958). Utilizing this method, the mass ratio of hesperidin to neohesperidin in Changshanhuyou was relatively quantified as 3.7:96.3 with RSD at 2.9%. Finally, using internal standard method, the absolute quantitative analysis was performed with acceptable reproducibility (RSD 1.3 and 4.5% for intra- and inter-day variations) and recoveries (from 95.9% to 108.9%), acceptable limit of detection (0.33 ng). In general, OLE–QTOF-MS/MS represented a promising and practical method for simple, rapid and effective analysis of isomeric compounds in complex matrices.