Flavonoid glycosides exist widely in medicine herbs and often used as nutraceuticals because of their excellent bioactivity and low toxicity. For accurate quality control and bioactivity assessment of Sphaerophysa salsula, a rapid and productive method to isolate flavonoid glycosides is needed. Therefore, this work reports the development of a novel comprehensive strategy based on an online middle-pressure chromatography and preparative HPLC for rapid enrichment and separation of flavonoid glycosides from Sphaerophysa salsula. First, the flavonoid glycosides were enriched using an online middle-pressure chromatographic column containing stationary MCI phase. During this process, the high-volume injection of the extracting solution was realized by an empty pre-column positioned before the main chromatographic tower. Then, the compounds were separated through preparative HPLC with Megress C18. As a result, one new flavonol 3-O-glycoside (2) together with two known flavonol 3-O-glycosides (1, 3) were targetedly isolated from Sphaerophysa salsula. The content of compounds 1-3 in Sphaerophysa salsula was 0.09, 0.11, and 0.18 wt%, respectively. Comparing to traditional enrichment and separation methods, our technique offers significantly shorter sample pretreatment time as well as high reproducibility. We believe that our separation method has a strong potential to be used for the processing of other medicinal plants. This article is protected by copyright. All rights reserved.
A novel chromatographic separation method for rapid enrichment and isolation of novel flavonoid glycosides from Sphaerophysa salsula