基于超高效合相色谱对黄芪中5种主要黄酮类化合物的快速检测  

Rapid Separation and Determination of 5 Kinds Flavonoids in Astragali Radix Based on Ultra-performance Convergence Chromatography

作　　者：王波[1,2] 周围 刘小花[1] 柳小亚 陈亚丽[1] 封士兰[1] 

机构地区：[1]兰州大学药学院,兰州730000 [2]甘肃出入境检验检疫局综合技术中心,兰州730000

出　　处：《分析化学》2016年第5期731-739,共9页Chinese Journal of Analytical Chemistry

基　　金：国家'十二五'科技支撑计划课题项目(No.2011BAI05B02)资助~~

摘　　要：建立了利用超高效合相色谱法(Ultra performance convergence chromatography,UPC^2)快速分离和测定黄芪中5种主要黄酮类化合物(毛蕊异黄酮、毛蕊异黄酮苷、美的紫檀素、芒柄花苷、芒柄花素)的方法。黄芪样品采用80%乙醇提取后,以超临界CO_2-0.2%H_3PO_4-甲醇溶液为流动相,梯度洗脱,色谱柱为Waters ACQUITY UPC^2CSH柱(100 mm×3.0 mm,1.8μm),柱温为40℃,流速为0.4 m L/min,进样量为1μL,检测波长为280 nm。整个分析过程仅需15 min,分析速度是传统液相色谱的3倍以上。结果表明:毛蕊异黄酮、毛蕊异黄酮苷、美的紫檀素、芒柄花苷与芒柄花素的检出限及定量限范围分别在0.3~0.5 mg/kg和1.0~2.0 mg/kg之间,5种黄酮类成分的加标平均回收率均高于99.7%,相对标准偏差(RSD)小于2.2%(n=6);在最优条件下,对13批不同产地的黄芪进行检测,毛蕊异黄酮、毛蕊异黄酮苷、美的紫檀素、芒柄花苷与芒柄花素的含量范围分别在4.8~102 mg/kg、14~277 mg/kg、0~135 mg/kg、5.3~119 mg/kg及2.8~41 mg/kg之间;本方法简便快速,重复性良好,结果准确可靠,可用于黄芪药材中5种主要黄酮类成分的含量测定。A method was developed for the rapid separation and determination of 5 kinds flavonoids （calyeosin, calycosin-7-O-β-D-glucoside, ononin, medicarpin and formononetin） in Astragali Radix based on ultra-performance convergence chromatography. After extracting by 80% ethanol, the flavonoids were separated on a Waters Acquity UPC2 CSH （ 100 mm×3.0 mmi. d. , 1.8 μm） column at 40℃ by using supercritical CO：-methanol （contain 0. 2% H3PQ ） acetonitrile as the mobile phase at a flow rate of 0.4 mL/min, and then analyzed by a UV detector at wavelength of 280 nm, the whole analysis progress was only 15 rain. The results showed that the limits of detection （LOD） and the limits of quantitation （LOQ） of five flavonoids were between 0.3 and 0. 5 mg/kg, and 1.0 and 2. 0 mg/kg, respectively. The spiked recoveries were more than 99.7%, and the relative standard deviations （RSD） were less than 2.2% （n=6）. Under the optimal conditions, 13 groups of Astragali Radix from different producing areas were detected. The contents of calycosin, calycosin-7-O-,8-D-glueoside, mediearpin, ononin and formononetin were 4.8-102 mg/kg, 14-277 mg/kg, 0-135 mg/kg, and 5.3-119 mg/kg, 2.8-41 mg/kg, respectively. This method is simple, fast, accurate and reproducible, and the result is reliable. The method is applicable for the determination of 5 tlavonoids in Astragali Radix.

关 键 词：超高效合相色谱 黄酮类 黄芪 

分 类 号：R284.1[医药卫生—中药学] O657.7[医药卫生—中医学]