The Determination Of Mainly Functional Compounds And Preliminary Construction Of The Metabolicprofiling Of Kunlun Chrysanthemum (Coreopsis Tinctoria)

By using the modified high performance liquid chromatography(HPLC)method,the content of chlorogenic acid,galuteolin,3,5-O-caffeoylquinic acid,and quercitrin were determined,and the extraction method of GC-MS samples were established in the different strains of Kunlun Chrysanthemum(Coreopsis tinctoria).The metabolic profiling of Kunlun Chrysanthemum was preliminary constructed through the research of the composition and content changes of metabolites,including amino acids organic acids,alcohols,sugar and others.Meanwhile according to the variable importance in the projection to the cluster pattern,the biomarkers were found which could be used as one of the evidences for the species identification and quality control of Kunlun Chrysanthemum.The results provide an analysis technology platform of Kunlun Chrysanthemum metabolic profiling and a basis for further studying on metabolism of ingredients.The main results were as follows:1.The suitable HPLC conditions were Alitima HP C18 5μm chromatographic column(250mm×4.6mm),acetonitril-0.1%phosphoric acid aqueous gradient elution,flow rate at 1 mL/min,detection wavelength at 350nm,column temperature 40℃,and sampling volume 5μL.2.The derivatization method for the pretreatment of Kunlun Chrysanthemum samples was preliminary constructed by GC-MS.In this study,oximation conditions of the samples were that the oxime time was 120min,temperature was 37℃,and the dosage of reagent(hydroxylamine hydrochloride)was 60μL.While derivatization conditions were that the derivatization time was 90min,temperature 37℃,and the dosage of reagent(99%BSTFA+1%TMCS)was 60μL.3.The methanol and water phase extract of Kunlun Chrysanthemum samples was mainly sugar and glucoside,organic acids,amino acids,etc,the chloroform phase extract was mainly organic acids,alkanes,etc.4.The biomarkers of the methanol and water phase extract from five species of Kunlun Chrysanthemum were as follows:(1)The five strains of Kunlun Chrysanthemum were divided into 2 groups using clustering analytical method,one group included Xue 1,Xue 4 and Xue 3,Xue 5,Xue 2 were attributed to another group,which indicuted that Xue 2 might have obvious different compounds compared with the others.(2)Alcohols,organic acids,alkanes were the most abundant component of Kunlun Chrysanthemum samples.Alanine,phenylalanine,methionine,leucine,glutamic acid and asparagine were the most abundant component of amino acids of Kunlun Chrysanthemum samples.Glucose,fructose and sucrose were the most abundant component of carbohydrate,galactose and maltose were the least abundant component.Undecane,octadecane,eicosane,octane were the most abundant component of alkanes.Succinate,oxalic acid,acidBenzoic acid were the most abundant component of organic acids.5.The metabolites from different strains of Kunlun Chrysanthemum were studied,theresults were that nine biomarkers were found respectively L-threonic acid,malonic acid,inositol,citric acid,fructose,sucrose,stearic acid,L-glutamine,y-aminobutyri.6.The metabolites from Kunlun Chrysanthemum and Florists Chrysanthemum were studied to find biomarkers,these biomarkers were docosanoic,octadecanoic acid,linoleic acid.The content of these biomarkers in Kunlun Chrysanthemum were higher than Florists Chrysanthemum.