| Summary: | <i>Polygonatum</i> species have been used as traditional medicines and functional foods in Asia and Europe since ancient times. In this study, a fast and simple method based on liquid chromatography coupled with electrospray ionization mass spectrometry (LC-ESI-MS) was developed to systematically analyze and identify the steroidal glycosides in four major <i>Polygonatum</i> species distributed in Japan, including <i>P. odoratum</i>, <i>P. falcatum</i>, <i>P. macranthum</i>, and <i>P. sibiricum</i>. As a result, 31 steroidal glycosides were tentatively identified, including 18 known and 13 previously unreported glycosides. Their structures were identified by the interpretation of chromatographic behavior and ESI-MS fragmentation patterns. The identification of 31 steroidal glycosides was indicative of a common biogenetic pathway in <i>Polygonatum</i> species. Our study disclosed the chemical profiling of steroidal glycosides in the plants of <i>Polygonatum</i> species, which will benefit better phytochemotaxonomical and phytochemical understanding and quality control for their medicinal usage.
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