黄芪中黄酮类化合物的超临界流体色谱分离方法研究  

Study on Separation of Flavonoids in Astragalus membranaceus by Supercritical Fluid Chromatography

作　　者：赵淑军 董姣姣 刘洁[2,3] 贾志鑫 闫晓宁 郁映婷 陈奕君 李月婷[1,2] 肖红斌 ZHAO Shu-jun;DONG Jiao-jiao;LIU Jie;JIA Zhi-xin;YAN Xiao-ning;YU Ying-ting;CHEN Yi-jun;LI Yue-ting;XIAO Hong-bin(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China;Research Center for Chinese Medicine Analysis and Transformation,Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院,北京100029 [2]北京中医药大学中药分析与转化研究中心,北京100029 [3]北京中医药大学北京中医药研究院,北京100029

出　　处：《分析测试学报》2021年第9期1311-1317,共7页Journal of Instrumental Analysis

基　　金：国家自然科学基金项目(81703947,81774155)。

摘　　要：考察了超临界流体色谱(SFC)中的色谱柱、改性剂、添加剂、流速、柱温和背压等因素对9种黄酮类成分(包括芒柄花素、异鼠李素、毛蕊异黄酮、山奈酚、槲皮素、紫云英苷、芒柄花苷、异槲皮苷、毛蕊异黄酮葡萄糖苷)分离的影响,与高效液相色谱法(HPLC)进行了比较,并建立了黄芪饮片中5种主要黄酮类化合物的SFC分析方法。采用Agilent ZORBAX RX-SIL色谱柱(4.6 mm×150 mm,5μm)进行分离,CO2-0.1%磷酸甲醇溶液为流动相梯度洗脱,流速为3 mL/min;柱温为35℃;背压为10 MPa,9种黄酮类化合物可在10 min内实现基线分离。5种黄酮类化合物在一定质量浓度范围内均具有良好的线性关系(r^(2)≥0.9632),检出限为10.69~16.21μg/mL,日内相对标准偏差(RSD)为1.3%~2.0%;日间RSD为1.6%~2.2%。5种黄酮类化合物在48 h内具有良好的稳定性,重复性为3.6%~6.0%,回收率为91.8%~112%。与HPLC法相比,9种化合物的保留时间顺序基本相反,SFC法更快速、经济环保,且其保留及选择性受色谱柱、改性剂和添加剂的影响较大,添加剂对色谱峰形影响明显。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,5μm)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(r^(2)≥0.9632)in the certain concentration ranges.The detection limits ranged from 10.69μg/mL to 16.21μg/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.

关 键 词：超临界流体色谱(SFC) 高效液相色谱 黄芪 黄酮类化合物 快速分析 

分 类 号：O657.7[理学—分析化学] R914.1[理学—化学]