Studies On The Chemical Constituents From Aster Tataricus Based On Metabolomics

Aster tataricus rhizoma,the dried rhizome of Aster tataricus L.f.,is one of the oldest traditional Chinese medicine in China.It was grown on the knap or wet land.It was widespread in China and mass cultivated in Hebei and Anhui Province.It had been used for relieving cough,moistening the lungs and preventing asthma for over 2000 years.In this study,HPLC-MS/MS for the quanitative analysis of constituents in Aster tataricus were developed.The chemical changes of Aster tataricus at different collection time were detected by using UPLC-TOF-MS.The possible difference of metabolic between dried Aster tataricus and after processing was determined by using GC-MS.And the different parts of Aster tataricus were also selected to compare metabolic changes using GC-MS.Part one Simultaneous determination of nine constituents in Astertataricus by HPLC-MS/MSObjective: To develop an HPLC-MS/MS method for the simultaneous determination of nine constituents(kaempferol,quercetin,luteolin,isorhamnetin,scopoletin,umbelliferone,ferulic acid,caffeic acid,and chlorogenic acid)in Aster tataricus.Methods: Analysis was performed on a Diamonsil C18 column(150 mm × 4.6 mm,5 ?m)eluted with acetonitrile(0.05% formic acid)and water(0.05% formic acid)in a gradient program.The flow rate was 0.8 mL/min,the injection volume was 10 ?L,and the column temperature was 30 ?.The multiple-reaction monitoring scanning(MRM)was employed for the quantification with switching electrospray ion source polarity in negative mode.The ion spray voltage was set at-4500 V and the turbo spray temperature was maintained at 650 ?.Results: Kaempferol,quercetin,luteolin,isorhamnetin,scopoletin,umbelliferone,ferulic acid,caffeic acid,and chlorogenic acid showed a good linearity in the ranges of 0.970-30.30 ?g/mL(r = 0.9987),0.498-15.55 ?g/mL(r = 0.9996),0.014-0.438 ?g/mL(r = 0.9995),0.022-21.95 ?g/mL(r = 0.9985),0.011-1.100 ?g/mL(r = 0.9995),0.008-0.247 ?g/mL(r = 0.9998),0.194-6.050 ?g/mL(r = 0.9996),0.197-6.150 ?g/mL(r = 0.9995),and 1.459-45.609 ?g/mL(r = 0.9990).The average recoveries varied from 97.43%-103.87%.The RSD of precision varied from 1.95%-3.09%.Conclusions: The method is simple,rapid,and sensitive.It could be used as a quantitative determination method for the nine components in Aster tataricus.Part two Difference study of Aster tataricus from different collectiontime by UPLC-Q-TOF-MS.Objective: To use an UPLC-Q-TOF-MS to evaluate the chemical difference of Aster tataricus from different collection time.Methods: The chromatographic separation was performed on an Agilent Poroshell 120 EC-C18 colume with water-acetonitrile as mobile phase in a gradient elution mode.The mass spectrometer equipped with electrospray ionization source was used as detector under the positive ion mode.The difference of Aster tataricus from different collection time were analysed by principal component analysis(PCA)with Markerview software.Results: Each group was separated from the other two groups successfully through the PCA,and four chemical metabolites were identificated.In February,the contents of quercetin,kaempferol,betulin were increased,and in November,the content of quercitrin was increased.Conclusions: From the metabonomic perspective,the chemical contents of Aster tataricus from different collection time were disparate.It also provided help for the collecting of Aster tataricus and clinical application.Part three Comparative metabolomics in different part and differentprocessed products of Aster tataricus by GC-TOF-MSObjective: Metabolomic analyses of different part and different processed products of Aster tataricus L.f.by using plant metabolomic technology based on GC-TOF-MS.Methods: Eight batches of samples(aerial part and underground part,Aster tataricus and Aster tataricus fried with honey)were determined by GC-TOF-MS.The results were used to PCA analysis and t test by using the SIMCA 14.1 software from Umea company.Results: The results showed that the chemical constituents were obviously different from the two samples(aerial part and underground part,Aster tataricus and Aster tataricus fried with honey).The content of amino acids and organic acids in underground part were higher than in aerial part.The content of organic acids in Aster tataricus were higher than in Aster tataricus fried with honey.After processing,the content of carbohydrates increased to some degree.At the same time,the contents of some amino acids,sugar alcohols and osamines were different.Conclusions: From the metabolomic perspective,the chemical contents of Aster tataricus from different part were disparate.Different part of Aster tataricus may change the medicinal properties so as to lead the various pharmacological activities.The chemical contents in Aster tataricus and Aster tataricus fried with honey were also very different.Processing may change the medicinal properties so as to lead the various pharmacological activities.