浊点萃取-高效液相色谱法同时测定槲寄生中3种黄酮的含量Determination of three flavonones in Viscum coloratum(Komar.) Nakai by HPLC-UV with cloud-point extraction
范荣华,赵云丽,苏畅,谢瑶,闫敬敬,于治国
FAN Rong-hua,ZHAO Yun-li,SU Chang,XIE Yao,YAN Jing-jing,YU Zhi-guo(School of Pharmacy
摘要(Abstract):
目的采用浊点萃取法富集槲寄生中的异鼠李素-3-O-β-D-葡萄糖苷、鼠李秦素-3-O-β-D-葡萄糖苷、鼠李秦素-3-O-β-D-(6″-β-羟基-β-甲基戊二酸半酯)葡萄糖苷,并使用高效液相色谱法测定其含量。方法采用聚乙二醇辛基苯基醚(Triton X-114)作为萃取剂,色谱柱为Synergi C18柱(250 mm×4.6 mm,4μm),流动相为乙腈(A)和体积分数0.5%乙酸溶液(B)梯度洗脱,流速为1.0 mL.min-1,柱温为30℃,检测波长为358 nm。结果测得异鼠李素-3-O-β-D-葡萄糖苷、鼠李秦素-3-O-β-D-葡萄糖苷、鼠李秦素-3-O-β-D-(6″-β-羟基-β-甲基戊二酸半酯)葡萄糖苷质量浓度分别在2.093~41.86、1.200~24.00、0.861 0~17.22 mg.L-1内与峰面积呈良好线性关系;平均回收率分别为93.2%、95.6%、94.5%。结论该方法准确、灵敏,可作为槲寄生药材质量控制的有效方法。
Objective To develop a method based on cloud-point extraction(CPE)for the determination of isorhamnetin-3-O-β-D-glucoside,rhamnazin-3-O-β-D-glucoside,rhamnazin-3-O-β-D-(6″-β-hydroxy-β-methylglutaryl)-glucoside in the Viscum coloratum(Komar.)Nakai by HPLC-UV.Methods The non-ionic surfactant Triton X-114 was chosen as the extract solvent and various experimental conditions were investigated to optimize the extraction process.Under optimum conditions,the extracts were analyzed by reverse-phase column.Acetonitrile and 0.5% acetic acid were used as the mobile phase under gradient conditions.The flow rate was 1.0 mL · min-1 and the UV detection wavelength was set at 358 nm.Results All calibration curves showed good linearity(r>0.999)within test ranges.The recoveries,measured at three concentration levels,varied from 93.2% to 95.6%.Conclusions The developed and validated methods are simple,accurate and can be selected as the techniques for separation and quantification of the three flavonones in the Viscum coloratum(Komar.)Nakai.
关键词(KeyWords):
浊点萃取;聚乙二醇辛基苯基醚;高效液相色谱法;黄酮;含量测定
cloud-point extraction;Triton X-114;HPLC-UV;flavonone;content determination
基金项目(Foundation): 国家自然科学基金资助项目(30901967);; 辽宁省博士科研启动基金资助项目(20091078);; 中国博士后科学基金资助项目(2011M500579)
作者(Author):
范荣华,赵云丽,苏畅,谢瑶,闫敬敬,于治国
FAN Rong-hua,ZHAO Yun-li,SU Chang,XIE Yao,YAN Jing-jing,YU Zhi-guo(School of Pharmacy
DOI: 10.14066/j.cnki.cn21-1349/r.2012.01.008
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