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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.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objectives<\/jats:title><jats:p>A non\u2010targeted liquid chromatography\u2010quadrupole time\u2010of\u2010flight mass spectrometry method coupled with an all ion fragmentation\u2010exact neutral loss (AIF\u2010ENL) strategy was developed for the screening of pyranosides in plants.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Pyranosides in various types were collected as a model. The AIF\u2010ENL strategy comprised three steps: AIF spectrum acquisition and generation, ENL\u2010based searching and identification, and confirmation of structural type using target second\u2010stage 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 <jats:italic>Rhodiola crenulata<\/jats:italic> (Hook. f. et Thoms) H. Ohba.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The method was proved to be an efficient tool for the screening of pyranosides. When it was applied to <jats:styled-content style=\"fixed-case\"><jats:italic>R. crenulata<\/jats:italic><\/jats:styled-content>, 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 <jats:styled-content style=\"fixed-case\"><jats:italic>R. crenulata<\/jats:italic><\/jats:styled-content> for the first time.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The method offers a new platform for discovering pyranosides. In addition, the developed non\u2010targeted 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.<\/jats:p><\/jats:sec>","DOI":"10.1002\/pca.3045","type":"journal-article","created":{"date-parts":[[2021,3,28]],"date-time":"2021-03-28T22:41:00Z","timestamp":1616971260000},"page":"1039-1050","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Non\u2010targeted screening of pyranosides in <i>Rhodiola crenulata<\/i> using an all ion fragmentation\u2010exact neutral loss strategy combined with liquid chromatography\u2010quadrupole time\u2010of\u2010flight mass spectrometry"],"prefix":"10.1002","volume":"32","author":[{"given":"Tian","family":"Tian","sequence":"first","affiliation":[{"name":"The Research Centre of Chiral Drugs, Innovation Research Institute of 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