Preparative isolation of maltol glycoside from <i>Dianthus superbus</i> and its anti-inflammatory activity <i>in vitro</i>

Dianthus superbus is a traditional Chinese medicine that is commonly utilized as a treatment for inflammation, pain, and immunological conditions. In this study, an anti-inflammatory maltol glycoside derived from Dianthus superbus was isolated for the first time via medium and high-pressure liquid chromatography, and at the same time, the in vitro anti-inflammatory activity of this maltol glycoside was preliminarily explored. Initially, crude samples of Dianthus superbus were preprocessed via MCI GEL® CHP20P and Spherical C18 medium-pressure chromatography, under the guidance of evaluation of in vitro anti-inflammatory activity. Fr44 was found to be the target fraction, and it was further isolated via two-dimensional reversed-phase/hydrophilic interaction liquid chromatography, yielding > 95% pure and was identified as tunicoside B. MTT assay, nitric oxide and nitric oxide synthase were used to evaluate the effects of tunicoside B on murine macrophage Raw264.7 by nitric oxide synthase assay kit, molecular docking, and western blotting. The results showed that tunicoside B did not affect the viability of cells and exhibited significant anti-inflammatory activity. As far as we know, this is the first report of tunicoside B from Dianthus superbus and the first study on the anti-inflammatory activity of tunicoside B. More importantly, the approach established in this study is expected to provide a theoretical basis for the separation and pharmacological activity study of maltol glycosides from other natural products.