Purification of spinosin from Ziziphi Spinosae Semen using macroporous resins followed by preparative high-performance liquid chromatography

As a well-known traditional Chinese medicine, Ziziphi Spinosae Semen has been used for treating anxiety and insomnia for a long time. Spinosin, the main active C-glycoside flavonoid in Ziziphi Spinosae Semen, has attracted much attention because of its many pharmacological activities including strong hypnotic effects, anxiolytic-like effects, and so on. In the present work, high-purity spinosin was separated from Ziziphi Spinosae Semen using the HPD-300 resin followed by preparative high-performance liquid chromatography. The adsorption kinetics curve of spinosin on the HPD-300 resin was studied and fitted well by the pseudo-second-order equation. The adsorption isotherms were also constructed and low temperature favored the adsorption reaction. The separation parameters were optimized using dynamic adsorption and desorption tests. After a one-run treatment with HPD-300 resin, the concentration of spinosin increased 11.8-fold from 0.99 to 11.7% with a recovery yield of 80.4%. Furthermore, the purity of spinosin could surpass above 98% after separation by preparative high-performance liquid chromatography and recrystallization with a recovery yield of 72.6%. The developed method was effective and suitable for the large-scale preparation of spinosin. Moreover, it was confirmed that HPD-300 resin could enable good selection for the enrichment of flavonoids from different plants.