双相萃取-液相色谱检测细胞培养液样品中的山银花黄酮类化合物  

A dual phase extraction combined with HPLC method was developed to detect Flos Lonicerae flavonoids in cell culture medium

作　　者：郭玉[1] 朱鹏[1] 刘刚[1] 姚旭[1] 凌杉村 丁岚[1] 李冲 

机构地区：[1]南华大学药物药理研究所湖南省分子靶标新药研究协同创新中心,湖南衡阳421001

出　　处：《应用化工》2018年第2期410-412,415,共4页Applied Chemical Industry

基　　金：湖南省教育厅优秀青年基金(15B211);湖南省大学生研究性学习和创新性实验计划项目(2015226)

摘　　要：乙醇-磷酸氢二钾双水相体系萃取细胞培养液中的山银花黄酮类化合物,用高效液相色谱法分析样品中的山银花黄酮类化合物的含量,色谱柱为C_(18),流动相为甲醇-水(50%∶50%v/v,用磷酸酸化至pH 4.0),流速1.0 m L/min,检测波长352 nm。结果表明,双水相萃取山银花黄酮上相乙醇,得到5个信号较强的化合物,其色谱行为与未经萃取的相同浓度山银花黄酮粗提物样品基本一致;萃取山银花黄酮粗提物细胞培养液样品获同样结果,表明乙醇-磷酸氢二钾双水相萃取结合液相色谱可检测细胞培养液中的山银花粗提物的黄酮化合物,该双水相能较好萃取并浓缩黄酮类化合物分配于上相乙醇中,有利于成分复杂、低浓度生物样品中待测化合物的分离、纯化、浓缩。The Flos Lonicerae flavonoids in cell culture mediumin was extracted by the ethanol-K_2HPO_4 two-phase conditions and its content was detected by HPLC with a C_(18) column. Samples were eluted using methanol/water( 50% ∶ 50% v/v) with phosphoric acid( apparent pH 4. 0) at a flow rate of 1. 0 m L/min and detected at 352 nm. Results showed that,with the two-phase system extracting flavonoids crude extract from Flos Lonicerae,chromatographic behavior of the 5 obvious signals from the upper phase sample were almost the same as the flavonoids crude extract samples. When the samples of flavonoids crude extract from Flos Loniceraein cell culture medium were extracted by the sameprocedure,test results were also basically the identical. All these suggest the ethanol-K_2HPO_4 two-phase system combined with HPLC can be determined the Flos Lonicerae flavonoids in cell culture medium samples and the two-phase system can be better extraction and concentration of flavonoids in its upper phase,which is beneficial to the separation,purification and concentration of the test compounds in the biological sample with low concentration and complex composition.

关 键 词：双水相体系 乙醇/磷酸氢二钾 生物样品 黄酮类化合物 

分 类 号：TQ028[化学工程]