Study On The Technology Of Separation And Purification Of Five Species Characteristic Components In Recipe Xi Hong Hua Formula

This experiment is a preliminary experimental study on the molecular mechanism in the treatment of coronary heart disease in the 2016 NSFC-Xinjiang joint fund project.The purpose of this study is to separate the group of characteristic chemical constituents of the five species from Xi Hong Hua Formula(XHHF),including Crocus sativus L,Nardostachys jatamansiDC,Dracocephalum moldavica L,Anchusa azurea and Lavandula angustifolia Mill.This study have explored the technology of the separation and preparation of the Traditional Chinese Medicines(TCMS),provided the theoretical basis for the research of the National Natural Science Foundation of China,and laid the material foundation of pharmacological activity in XHHF.High-performance liquid chromatography(HPLC)is used to analyze the chemical constituents of five species from XHHF.Based on the chromatogram and literature information,it is deduced that the characteristic chemical composition group of 5 species are represented by crocin,crocetin,safranal and nardosinone,chlorogenic acid and rosmarinic acid,tilianin and daucosterol,respectively.HPLC and Thin Layer Chromatography(TLC)as detection means.Then screening the separation conditions of the extracts that extracted by five plants with 70% ethanol,concentrated and dried.The TLC system analysis method was used to screen the six commonly used organic solvents based on n-hexane,petroleum ether,ethyl acetate,chloroform,acetone,methanol,and n-butanol as the chromatographic separation packing materials and chromatographic separation of mobile phases.Then the best optimum mobile phase for the elution of five species that is confirmed by the number of spots appearing on the thin plate and the resolution is determined to be n-butanol-methanol-water,Chloroform-Acetone,Chloroform-Methanol.The best separation media is Silica Gel.Separation of five plants is performed using vacuum liquid chromatography(VLC),and six effective components were isolated from each plant.The characteristic components separated by VLC are analyzed by TLC and HPLC.The analysis results show that the separated components have a certain linear relationship and the process of the separation is reproducible.In summary,through the systematic study of the components from the five plants in the XHHF,a system process of the separation and preparation based on chromatographic techniques is established,which will be a systematic separation method of the TCMS and provide a reference of the pharmacological activities of the chemical ingredients in the future.