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

Tongyu Wei; Hanwen Ni

doi:10.1016/j.jchromb.2024.124141

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

J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Apr 26:1240:124141. doi: 10.1016/j.jchromb.2024.124141. Online ahead of print.

Authors

Tongyu Wei¹, Hanwen Ni²

Affiliations

¹ College of Resources and Environment Sciences, China Agricultural University, Beijing 100193, PR China.
² College of Resources and Environment Sciences, China Agricultural University, Beijing 100193, PR China. Electronic address: hanwenni@cau.edu.cn.

PMID: 38691943
DOI: 10.1016/j.jchromb.2024.124141

Abstract

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 %.

Keywords: Aqueous two-phase flotation; Astragalin; Flaveria bidentis; Preparative high performance liquid chromatography.