Metabonomics Strategy To Analyzing The Differential Metabolites In Propolis

Propolis,known as "purple gold",contains more flavonoids and phenolic acids,so it has a wide range of biological activities such as anti-oxidant,anti-inflammatory,anti-cancer,anti-radiation,anti-bacterial and so on.However,because of its low output and long production cycle,propolis market supply falls short of demand.In addition,the composition of propolis is complex and similar to that of poplar bud,which makes its adulteration in an endless stream.With the advancement of propolis commercialization,the lawbreakers often disguise poplar bud as propolis and sell them.In many previous studies,the determination of active ingredients such as phenolic acid and flavonoids in propolis was mainly based on national standards,or or targeted determination of a single compound method..This study is based on metabonomics technology,using liquid-mass spectrometry to analyze propolis comprehensively,and combining with multivariate statistical analysis to find the differential metabolites between propolis and propolis products from different origins.At the same time,the determination of metal pollutants in propolis provides a theoretical basis for the research of propolis and its products.This experiment is divided into two parts.The first part is the metabonomics study of propolis,the original data of propolis and propolis products from different origins are obtained by UPLC-Q Exactive Obitrap LC-MS.After that,we import the data into the Compound Discoverer2.1 software to extract the characteristic peaks in the total ion flow diagram of the sample,and combine the multivariate statistical analysis to distinguish the different samples to find possible differential metabolites.Then we use the secondary data obtained by high-resolution mass spectrometry,combined with combined with the data processing of Compound Discoverer2.1 software,to carry out preliminary characterization of metabolites;The second part is to detect metal contaminants in propolis samples.ICP-MS was used to determinethe aluminum,manganese,lead,cadmium,chromium,arsenic,iron and other elements in propolis samples,which provides a theoretical basis for the rational use of propolis products.The first part of the results shows that different propolis samples can be well distinguished.Between Shandong and Sichuan raw propolis,16 differential metabolites were found.Shandong raw propolis contains more Ortho-phthalaldehyde,isoliquiritigenin,3-Methoxy-5,7,3',4'-tetrahyd roxy-flavone,Tetramethylscutellarein and other substances,Ethyl vanillin propylene glycol acetal,Ethyl vanillin isobutyrate,Glabrol are mostly distributed in Sichuan raw propolis.A total of 16 differential metabolites were found in propolis from Sichuan and Hebei province.Sichuan propolis contains more Ginkgolic acid,Anacardic acid,18-?-Glycyrrhetinic acid and other substances,Hebei raw propolis contains more(+)-Marmesin,2-Hydroxyhippuric acid,Lovastatin and other substances.The differences between propolis from Shandong and Hebei province are relatively small.And a total of 11 differential metabolites were found,Shandong raw propolis contains Ginkgolic acid and Icomucret,raw propolis from Hebei contains more substances such as Pinocembrin,Sakuranetin,Isoliquiritigenin and so on.The reason for the differences in the raw propolis from three provinces may be the differences in the resin composition of the plant source of the propolis collected from the ground,supplemented by the impact of the environment and climate.The differences between raw propolis from three provinces and online propolis are small,and finally 11 kinds of differential metabolites are identified.There are more coumarins such as Scoparone,Phellopterin,and flavonoids such as hydroxyflavanone,Pinocembrin,3-Methoxy-5,7,3',4'-tetrahydroxy-flavone and so on.There are more contents of caffeic acid,Byakangelicol and Fuscaxanthone C in raw propolis.The reason for the difference is that beekeepers have less processing of propolis,and there are certain differences in the composition of propolis from different places.There were significant differences between raw propolis and propolis capsules.A total of 19 kind of differential metabolites were found,including Ginkgolic acid,chlorogenic acid,Cinnamic acid,Caffeic acid,Ferulic acid,Isoliquiritigenin,Tetramethylscutellarein and so on.Among them,the propolis capsules contains more Shikalkin,Tetramethylscutellarein,beta,beta-Dimethylacrylshikonin,[3,4,6-Triacetoxy-5-(benzoyloxy)-1-cyclohexen-1-yl]methyl benzoate,and the remaining 14 substances are more in raw propolis.The reason for the difference is the partial loss of ingredients in the processing of propolis and the addition of processing accessories.The second part of the results showed that the contents of Al,Cr,Fe,Ba and Pb in propolis were higher than the standard,and the contents of other elements such as V,Mn and Co were lower.The content of metal elements in 10 kinds of capsule products purchased at random did not exceed the national standard,and the metal pollution was small.