Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1306, Issue -, Pages 12-19Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2013.07.052
Keywords
Dynamic ultrasonic-assisted and solid-phase extraction; Ionic liquids; HSCCC; Rhodiola rosea; Flavonoids
Funding
- Jiangnan University Science Research Projects [JUSRP211A32]
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Purification of four flavonoids from Rhodiola rosea was developed by on-line combination of sample preparation and counter-current chromatography (CCC). Flavonoid sample was prepared by dynamic ultrasonic-assisted and solid-phase extraction using ion liquids as extractant. The preparation conditions were optimized by D-optimal design as follows: 2 mol/L of 1-ethyl-3-methylimidazolium bromide concentration, 360W of ultrasonic power, 1.5 mL/min of flow rate, 35 min of extraction time and 0.5 mL (absorbent) per g (material) of absorbent amount. The prepared sample solution (20 mL) was loaded and injected directly into CCC column for final separation. As a result, four flavonoids, herbacetin-3-O-beta-D-glucopyranosyl-7-O-alpha-L-rhamnopyranoside 1 (40.1 mg), kaempferol-3-O-beta-D-glucopyranosyl-7-O-alpha-L-rhamn-opyranoside 2 (4.6 mg), kaempferol 3-O-beta-D-glucopyranoside-(2 -> 1)-beta-D-xylopyranoside 3 (20.2 mg) and herbacetin-8-O-beta-D-glucopyranoside 4 (22.5 mg), were obtained from 20 g of R. rosea material using ethyl acetate-n-butanol-H2O as solvent system at a ratio of 4:1:5 by CCC. Their structures were identified by ESI-MS/MS, NMR methods. Their purities determined by UPLC were 98.5%, 95.4%, 98.1% and 97.5%, respectively. Kaempferol-3-O-beta-D-glucopyranosyl-7-O-alpha-L-rhamnopyranoside 2 and herbacetin-8-O-beta-D-glucopyrano-side 4 were isolated for first time from R. rosea. The purification method was simple, efficient and evaded tedious sample preparation process. (C) 2013 Elsevier B.V. All rights reserved.
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