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A silica gel orthogonal high-performance liquid chromatography method for the analyses of steviol glycosides: novel tetra-glucopyranosyl steviol

Perera, Wilmer H., Ramsaroop, Taylor, Carvalho, Raquel, Rodenburg, Douglas L., McChesney, James D.
Natural product research 2019 v.33 no.13 pp. 1876-1884
Stevia rebaudiana, acetonitrile, dissociation, diterpenoids, glycosides, high performance liquid chromatography, leaf extracts, nuclear magnetic resonance spectroscopy, purification methods, silica gel, steviol, tandem mass spectrometry
A silica gel orthogonal method using acetonitrile: water was developed for the analyses of fractions rich in very polar steviol glycosides and resolve regions of co-elution of these compounds in reversed-phase. Additionally, we also used this normal phase analytical method to scale up the purification process of steviol glycosides. Using these approaches, one novel minor tetra-glucopyranosyl diterpene glycosides together with three known compounds were purified from a commercial Stevia rebaudiana leaf extract. Compound 1 was unambiguously elucidated as 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]ent-kaur-16-en-19-oic acid-(6-O-β-D-glucopyranosyl-β-D-glucopyranosyl) ester (rebaudioside Y) based on high-performance liquid chromatography retention times, tandem mass spectrometry dissociation pattern and 1D and 2D NMR experiments. Known compounds were isolated in gram quantities and identified as rebaudioside D, E and M.