Preparative separation of mangiferin glycosides by high speed counter current chromatography and comparison of their antioxidant and antitumor activities

Mangiferin, a xanthonoid with various bioactivities. The low solubility of mangiferin limits the use in pharmacological fields. In this study, high-speed counter-current chromatography (HSCCC) was used to separate and purify mangiferin glycosides from the crude sample after enzymatic glycosylation of mangiferin. Two fructosyl mangiferin were successfully purified by HSCCC with a two-phase-solvent system composed of n-butanol–methanol–water (6 : 1 : 6, v/v). A total of 18 mg of mangiferin (I), 73 mg of β-d-fructofuranosyl-(2 → 6)-mangiferin (II), and 58 mg of β-d-difructofuranosyl-(2 → 6)-mangiferin (III) were obtained in one-step separation from 150 mg of the crude sample with purities of 99.2%, 98.7% and 98.9%, respectively. The chemical structures were identified by HRMS, (1)H-NMR, (13)C-NMR and 2D NMR. Mangiferin glycosides showed higher antioxidant and antitumor activities compared to that of mangiferin by employing DPPH scavenging effect, reducing power and cytotoxicity assay. Therefore, these novel fructosyl mangiferin exhibit a great potential to be developed into new medicines.