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| Study on Separation of Flavonoids in Astragalus membranaceus by Supercritical Fluid Chromatography |
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| DOI： |
| KeyWord:supercritical fluid chromatography（SFC） high performance liquid chromatography Astragalus membranaceus flavonoids rapid analysis |
| Author | Institution |
| ZHAO Shu-jun，DONG Jiao-jiao，LIU Jie，JIA Zhixin，YAN Xiao-ning，YU Ying-ting，CHEN Yi-jun，LI Yue-ting，XIAO Hong-bin |
1. School of Chinese Materia Medica， Beijing University of Chinese Medicine， Beijing ， China;2. Research Center for Chinese Medicine Analysis and Transformation， Beijing University of Chinese Medicine， Beijing ， China;3. Beijing Research Institute of Chinese Medicine， Beijing University of Chinese Medicine， Beijing ， China |
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| Abstract: |
| The effects of different chromatographic columns， modifiers， additives， flow rates， column temperatures and back pressures in supercritical fluid chromatography（SFC） on the separation of 9 flavonoids， e.g. formononetin， isorhamnetin， calycosin， kaempferol， quercetin， astragalin， ononin， isoquercitrin and calycosin-7-glucoside were investigated. Meanwhile， a SFC method was established for the determination of 5 flavonoids in Astragalus membranaceus， which was compared with high performance liquid chromatography（HPLC）. The separation was performed on an Agilent ZORBAX RX－SIL column（4.6 mm × 150 mm， 5 μm） by gradient elution， with CO2－0.1% phosphate acid methanol solution as mobile phase at a flow rate of 3 mL/min， a column temperature of 35 ℃ and a back pressure of 10 MPa. Results indicated that the nine flavonoids could be separated within 10 min. The five quantified flavonoids all had good linear relationships（r2 ≥ 0.963 2） in the certain concentration ranges. The detection limits ranged from 10.69 μg/mL to 16.21 μg/mL. The intra-day and inter-day relative standard deviations（RSD）were in the range of 1.3%－2.0% and 1.6%－2.2%， respectively. The five flavonoids had good stability within 48 h， with RSDs of 1.6%－3.8%. The repeatabilities of five flavonoids ranged from 3.6% to 6.0%. The average recoveries were in the range of 91.8%－112%. Compared with the HPLC method， the SFC method was more rapid， economical and environmental friendly as the retention time sequences of the components in the two methods were almost opposite. Its retention and selectivity were greatly affected by columns， modifiers and additives， and additives had obvious effects on chromatographic peak shape. |
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