Efficient new method for extraction and isolation of three flavonoids from Patrinia villosa Juss. by supercritical fluid extraction and high-speed counter-current chromatography

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• 作者：Jinyong Peng ; Guorong Fan ; Yifeng Chai and Yutian Wu
• 关键词：Patrinia villosa ; Counter-current chromatography ; SFE ; Flavonoid
• 刊名：Journal of Chromatography A
• 出版年：2006
• 期刊代码：47_00219673
• 类别：ch
• 出版时间：13 January 2006
• 卷：1102
• 期：1-2
• 页码：44-50
• 文件大小：251 K

摘要

Supercritical fluid extraction (SFE) of orotinin, orotinin-5-methyl ether and licoagrochalcone B from Patrinia villosa was performed. The optimization of parameters including pressure, temperature, modifier and sample particle size on yield was carried out using an analytical-scale SFE system. The process was then scaled up by 100 times using a preparative SFE system under the optimized conditions of 25 MPa, 45 °C, a sample particle size 40–60 mesh and modified CO₂ with 20%methanol. The yield of the preparative SFE was 2.82%(crude extract I) and the combined yield of orotinin, orotinin-5-methyl ether and licoagrochalcone B was 0.82 mg/g of dry sample mass. Then the crude extract I was re-dissolved in methanol and methanol soluble fraction (crude extract II, 0.17%) was obtained, which was successfully isolated and separated by a preparative high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane–ethyl acetate–methanol–water (5:6:6:6, v/v/v/v) by increasing the flow-rate of the mobile phase stepwise from 1.0 to 2.0 ml/min after 3 h. The target compounds isolated and purified by HSCCC were analyzed by high performance liquid chromatography. The separation produced total of 38.2 mg of orotinin at 99.2%purity, 19.8 mg of orotinin-5-methyl ether at 98.5%purity and 21.5 mg of licoagrochalcone B at 97.6%purity from 400 mg of the crude extract in a one-step separation. The recoveries of orotinin, orotinin-5-methyl ether and licoagrochalcone B were 91.1, 91.6 and 90.3%, respectively, and the chemical structure identification was carried out by UV, IR, MS, ¹H NMR and ¹³C NMR.