Non‐targeted screening of pyranosides in Rhodiola crenulata using an all ion fragmentation‐exact neutral loss strategy combined with liquid chromatography‐quadrupole time‐of‐flight mass spectrometry

Abstract Introduction Pyranosides as one kind of natural glycosides contain a pyran ring linked to an aglycone in the structure. They occur widely in plants and possess diverse biological activities. The discovery of new pyranosides not only contributes to research on natural products but also may promote pharmaceutical development. Objectives A non‐targeted liquid chromatography‐quadrupole time‐of‐flight mass spectrometry method coupled with an all ion fragmentation‐exact neutral loss (AIF‐ENL) strategy was developed for the screening of pyranosides in plants. Methods Pyranosides in various types were collected as a model. The AIF‐ENL strategy comprised three steps: AIF spectrum acquisition and generation, ENL‐based searching and identification, and confirmation of structural type using target second‐stage mass spectrometry (MS/MS). The strategy was systematically evaluated based on the matrix effects, fragmentation stability, scan rate and screening efficiency and finally applied to Rhodiola crenulata (Hook. f. et Thoms) H. Ohba. Results The method was proved to be an efficient tool for the screening of pyranosides. When it was applied to R. crenulata , a total of 24 pyranoside candidates were detected. Among them, six were tentatively identified on the basis of the agreement of their elemental composition with the reported. The other 18 were detected in R. crenulata for the first time. Conclusion The method offers a new platform for discovering pyranosides. In addition, the developed non‐targeted strategy can also be used for other natural products, such as flavonoids and coumarins, as long as there is a common fragmentation behaviour in their MS/MS to generate characteristic neutral losses or fragments.