高聚物型色谱柱在高效液相色谱法中一步分析制备葛根素及大豆苷元的应用  

One-step determination and preparation of puerarin and daidzein by polymer column in HPLC

作　　者：夏明[1] 刘晓宁[1] 魏荣卿[1] 郑涛[2] 

机构地区：[1]南京工业大学生物与制药工程学院,江苏南京211800 [2]中国科学院广州能源研究所,广东广州510640

出　　处：《生物加工过程》2017年第1期37-42,共6页Chinese Journal of Bioprocess Engineering

基　　金：国家重点基础研究发展计划(973计划)(2013CB733504);国家高技术研究发展计划(863计划)(2012AA021203)

摘　　要：建立了使用以聚苯乙烯-二乙烯基苯(PS-DVB)为基质的高聚物型色谱柱一步分离制备葛根提取液中的葛根素和大豆苷元的方法。采用MKF-RP-HH色谱柱(300 mm×7.8 mm,8μm),葛根提取液经70%乙醇溶解稀释,以水(A)和甲醇(B)为流动相进行梯度洗脱,流速1.0 m L/min,检测波长250 nm,柱温30℃。结果表明:葛根提取液中的2种有效成分葛根素和大豆苷元均与杂质达到了较好的分离效果,且柱效大于2 100 N/m,优于常规的C-18硅胶色谱柱(柱效:1 000 N/m)。该高聚物型色谱柱不仅可用于上述分析,还可用作半制备柱制备得葛根素和大豆苷元纯品,葛根素在2~50μg/m L范围内线性关系良好,回归方程为y葛根=1.342 2x-1.018 4,线性相关系数为0.999;大豆苷元在2~20μg/m L范围内线性关系良好,回归方程为y大豆=2.275x+0.869 5,线性相关系数为0.999;葛根素和大豆苷元的纯化得率分别可达95.9%和78.5%。Polymeric chromatographic column with polystyrene-divinyl-benzene （PS-DVB） packing materials （MKF-RP-HH,300 mm×7. 8 mm,8μm） was established to determinate and prepare puerarin and daidzein.The kudzu extraction dissolved and diluted by 70% ethanol.Water （A） /me tha nol （B） was served as mobile phase at the rate of 1.0 mL/min.Detection was done at the wavelength of 250 nm at 30 ℃.Two active ingredients （ puerarin and daidz ein ） and impurities have achieved effective separation. Column efficiency reached 2 100 N/m that is superior than conventional silica column （ 1 000 N/m） .This polymeric column can be used as semi-preparative column to prepare puerarin and daidzein.The selected chromatographic conditions led to effectively determinating puerarin with good linearity within 2-50 pg/mL,the regression equation wasy （puerarin ） = 1.342 2x-1. 018 4,and the regression coefficient was 0.999; the selected chromatographic conditions were found to effectively determinate daidzein with good linearity within 2-20 pg/mL,the regression equation was y （ daidzein ） = 2.275x ＋ 0. 8695 , and the regression coefficient was 0. 999.The purification efficiency ol puerain and daidzein could reach to 95. 9% and 78.5%.

关 键 词：聚苯乙烯二乙烯基苯 高聚物型色谱柱 反相高效液相色谱法 葛根素 大豆苷元 

分 类 号：O65[理学—分析化学] TQ464.1[理学—化学]