低共熔溶剂萃取高效液相色谱法测定紫草中的活性成分

Determination of active components in arnebia euchroma by deep eutectic solventa extraction high performance liquid chromatography

建立了一种基于疏水性低共熔溶剂涡旋辅助分散液液微萃取法用于提取紫草中的乙酰紫草素和β,β′-二甲基丙烯酰紫草素,并结合高效液相色谱法测定对目标分析物进行分离与测定。本研究以四丁基氯化铵为氢键受体,正辛酸为氢键供体,合成了一种新型疏水性低共熔溶剂作为萃取剂,其在涡旋辅助作用下分散于样品水溶液中,以增加萃取剂与目标分析物的接触面积,经离心后疏水性低共熔溶剂与样液实现相分离,吸取低共熔溶剂相经乙腈稀释后过滤,进行色谱分析。实验结果表明,在线性范围内各目标分析物具有良好的线性关系(R≥0.9999),其检出限和定量限的范围分别为0.31~0.49μg/L和1.03~1.64μg/L。日内精密度和日间精密度分别为0.33%~1.87%和0.87%~2.60%。本方法以疏水低共熔溶剂代替传统有机溶剂作为萃取剂,具有检出限低、萃取时间短、成本低、绿色环保等特点,可应用于紫草中活性成分的测定。

A liquid-liquid microextraction method based on hydrophobic deep eutectic solvents vortex assisted dispersion was established to extract acetyl shikonin andβ,β′-dimethylacrylic shikonin from Arnebia euchroma,and the target analytes were separated and determined by HPLC.In this study,tetrabutylammonium chloride was used as hydrogen bond acceptor and n-octanoic acid was used as hydrogen bond donor,and a new hydrophobic deep eutectic solvents was synthesized as extractant.It was dispersed in the sample aqueous solution with the aid of vortex to increase the contact area between the extractant and the target analyte.After centrifugation,hydrophobic deep eutectic solvents was separated from the sample solution,and the deep eutectic solvents phase was diluted with acetonitrile and filtered for chromatographic analysis.The experimental results show that the target analytes have a good linear relationship(R≥0.9999)in the linear range,and the limits of detection and quantification are 0.31~0.49μg/L and 1.03~1.64μg/L,respectively.The intra-day precision and inter-day precision were 0.33%~1.87%and 0.87%~2.60%respectively.This method uses hydrophobic deep eutectic solvents instead of traditional organic solvent as extractant,which has the characteristics of low detection limit,short extraction time,low cost,environmental protection and so on,and can be applied to the determination of active components in Arnebia euchroma.