ABSTRACT
A high-speed counter-current chromatography (HSCCC) method was successfully developed for the preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus in one step. The purity of deoxyschizandrin was 98.5%, and the structure was identified by MS, UV and NMR. This method was simple, fast, convenient and appropriate to prepare pure compound as reference substances for related research on Schisandrae Sphenantherae Fructus.
ABSTRACT
An efficient method for the isolation and purification of 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis by high speed counter-current chromatography (HSCCC) was established in this paper.The ether extracts of Radix Eupatorii Chinensis were purified by HSCCC with a solvent system of hexyl hydride ethyl acetate-methanol-water (1∶2∶1∶2,v/v/v/v).The upper phase was used as the stationary phase and the lower phase as the mobile phase.About 8.4 mg of 12,13-dihydroxyeuparin was obtained from 200 mg of ether extracts from Radix Eupatorii Chinensis in one-step HSCCC separation,with the purity of 96.71％,as determined by HPLC.After methanolwater recrystallization,the purity of 12,13-dihydroxyeuparin reached 99.83％.Such a simple and effective method was fairly useful to prepare pure compound as reference substances for related study on Radix Eupatorii Chinensis.
ABSTRACT
A reversed phase high performance liquid chromatography （HPLC） method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 column （250 mm × 4.6 mm, 5 μm） was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid （0.2%, v/v）. Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃. An ultraviolet （UV） detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12, 13-dihydroxyeuparin （r〉0.9999） and 106.9-1068.9μg/mL for glycyrrhizic acid （r〉0.9999）, respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13- dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.
ABSTRACT
A reversed phase high performance liquid chromatography (HPLC) method was established for the simultaneous determination of 12,13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.A Grace Apollo C18 column (250 mm× 4.6 mm,5 μm) was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid (0.2％,v/v).Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃.An ultraviolet (UV) detector was used with a selected wavelength of 240 nm.Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12,13-dihydroxyeuparin (r＞0.9999) and 106.9-1068.9μg/mL for glycyrrhizic acid (r＞0.9999),respectively.Recoveries were 102.18 ％ for 12,13-dihydroxyeuparin and 101.17％ for glycyrrhizic acid.The method developed could be applied to the simultaneous determination of 12,13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.