Studies In Quality Control And Evaluation Method Of Menthae Haplocalycis Herba Based On The Correlation Between Its Chromatographic Fingerprint And Antioxidant Activity

With more and more negative reports of synthetic antioxidants, the natural plants with antioxidant activity have became the focus of research. Menthae haplocalycis Herba, as a medicinal and edible plant, has good antioxidant capacity. Therefore, this paper aims to improve the quality control and evaluation method of Menthae haplocalycis Herba based on the correlation between its chromatographic fingerprint and antioxidant activity.Summary of the related literatureStudies on relationship of chromatographic fingerprint and pharmaceutical activitives in recent years were reviewed. Ninty references were cited. Part one:Correlation between chromatographic fingerprint and antioxidant activity ofMenthae Haplocalycis HerbaThe characteristic fingerprints of35batch of mint were determined by the method we optimized. Meanwhile, we determined the antioxidant activities of these samples by DPPH and FRAP assay respectively. Then we screened variables by PLS and obtained X5, X6, X12, X15, X19, X23which made great contributions to the DPPH scavenging activity; X4, X6, X8, X10, X11, X22, X24which made great contributions to the total antioxidant activity. It was found that these compounds were flavonoids and phenolic acids based on their ultraviolet absorption. Compared their retention time, we even found that X5and X4, X12and X8, X15and X11may be the same compound.Part two:Identification of compounds from Menthae Haplocalycis Herba by HPLC-MS/MSIn this chapter, we obtained38compounds based on their retention times and structures, also compared their Ms data with literature. Hesperetin, Hesperidin, Rutin, Diosmetin, Hispidulin, Diosmin, Acacetin, Genkwanin, Thymusin, Didymin, Luteolin-7-O-rutinoside, Vicenin-2, Linarin, Naringin, Apigenin-7-O-rutinoside, Xanthomicrol, Nevadensin,5,6dihydroxy-7,3’,4’-trimethoxy flavone,5,6-dihydroxy-7,8,3’,4’-tetramethoxy-flavone,5-hydroxy-6,7,3’,4’-tetramethoxy flavone, gardenin B, Caffeic acid,2-Propenoic acid,3(2,3,4-trihydroxyphenyl), Rosmarinic acid, Methyl rosemarinate, Lithospermic acid, trans Salvianolic acid J, Salvianolic acid B, Methylene tanshinquinone, Tanshinone Ⅰ, Tanshinone Ⅱ A, Dihydrogen tanshinone I or1,2-dihydro tanshinquinone, Oleanolic acid, Ursolic acid, Thymonin and its isomer, Luteolin-7-O-glucuronide and its isomer.Part three:Establishment and application of the determination method of Multi-target constitutions in Menthae Haplocalycis HerbaBased on the results mentioned before, we selected7target compounds to establish the quantitative method of mint. This chapter mainly include4parts.(1) Study in simultaneous determination of4flavonoids in Menthae Haplocalycis Herba by QAMS.(2) Combined with stoichiometry to study differences in compound contain of4flabonoids between2source areas、3harvest years.(3) The establishment and application of determination method of3phenolic acids.(4) Combined with stoichiometry to study differences in compound contain of3phenolic acids between2source areas、3harvest years.Prat four: HPLC specific chromatogram combined with stoichiometry for quality assessment of Menthae Haplocalycis HerbaTo analyze the HPLC specific chromatogram of different samples, this thesis used similarity evaluation, hierarchy cluster, principal component analysis successively. As a result, we reduce the data dimension and find the principle compound in classification.Part five:Establishment and application of quality control method of Menthae Haplocalycis Herba based on antioxidant activityWe established and used the optimum determination method of antioxidant activity by DPPH and FRAP assay respectively. Combined with the cluster results, we discuss the difficiency of the current quality standard.