Establishing aqueous two-phase flotation coupled with preparative high performance liquid chromatography and its application for the purification of astragalin from Flaveria bidentis

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 1240; p. 124141
• Main Authors: Wei, Tongyu, Ni, Hanwen
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-06-2024
• Subjects: Aqueous two-phase flotation; Astragalin; Flaveria bidentis; Preparative high performance liquid chromatography; Preparative high performance liquid chromatography; Astragalin; Flaveria bidentis; Aqueous two-phase flotation
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2024.124141

•ATFP coupled with prep-HPLC for purification of astragalin was established.•Recovery of astragalin from Flaveria bidentis was 99.5% using the ATFP method.•Using the established ATFP coupled with prep-HPLC, the purity of astragalin was 98.8%.
The strategy of aqueous two-phase flotation (ATPF) followed by preparative high performance liquid chromatography (prep-HPLC) was established and used for the separation of astragalin from Flaveria bidentis. In the ATPF, the effects of sublation solvent, solution pH, (NH4)2SO4 concentration in aqueous solution, cosolvent, N2 flow rate, flotation time and volumes of the PEG phase on the recovery of astragalin were investigated in detail, and the optimal conditions of ATPF were selected: 50 wt% PEG1000 ethanol solvent as the flotation solvent, pH 4, 350 g/L of (NH4)2SO4 concentration in 5 % ethanol aqueous phase, 40 mL/min of N2 flow rate, 30 min of flotation time, 10.0 mL of flotation solvent volume and twice. After ATPF enrichment, the flotation product was further purified by prep-HPLC. As determined by HPLC, the purity of astragalin was 98.8 %.