Metabonomics Study And Preliminary Construction Of Metabolites Database Of Pine Pollen

Pine pollen is rich in nutrients,including protein,polysaccharide,amino acid and other substances.It is known as complete nutrition.Will pine pollen affect its nutritional components during storage and sterilization? Are there differences in pine pollen treated by different storage periods and sterilization methods? In many studies,the determination of nutrients in pine pollen is based on national standards or other methods to determine a single substance.From the point of view of metabonomics,the metabolites of pine pollen were comprehensively analyzed by liquid continuum technology,and the differential metabolites of pine pollen were found by multivariate statistical analysis in different areas,different storage periods and different sterilization methods.At the same time,the differences of metabolites were studied by comparing other fresh pine flowers.Finally,a statistical analysis of the metabolites of fresh pine pollen was made,and a database of the metabolites of pine pollen was preliminarily constructed.The first part is the metabonomics of pine pollen.Fresh Yunnan pine pollen(YNS),fresh Pinus tabulaeformis pollen(YS),irradiated Pinus yunnanensis pollen(IR),ultra-high temperature instantaneous sterilization of Pinus yunnanensis pollen(UHT),one-year Pinus yunnanensis pollen(OY)and two-year Pinus tabulaeformis pollen(TY)were obtained by ultra-high performance liquid chromatography tandem four-bar orbital ion trap high resolution mass spectrometry(UHPLC-Q Exactive Obitrap LC-MS).The original data of water extracts of pollen and pine pollen were imported into Compound Discoverer software to extract the total ion flow graph chromatographic peaks of the samples,and multivariate statistical analysis was used to distinguish the different samples to find possible differential metabolites.The second part is the preliminary construction of pine pollen metabolites database.Similarly,fresh Yunnan pine pollen were detected by liquid chromatography.Compound Discoverer software was used to preliminarily process the original data,and then combined with secondary mass spectrometry information to achieve the preliminary qualitative analysis of compounds.The first part of the results show that the experiments can be well distinguished between different samples.14 kinds of differential metabolites were found between fresh Pinus yunnanensis pollen and Pinus tabulaeformis pollen.Pinus yunnanensis pollen contains more kaempferol,N-(2-hydroxypropyl)methacrylamide,androsterol,tyramine,etc.The differential metabolites distributed in the pollen of Pinus tabulaeformis pollen are abietic acid,methyl ricinoleate,oleoylglycine and the like.Irradiation sterilization and ultra-high temperature instantaneous sterilization of pine pollen found 11 different metabolites,including(1Z,5Z)-1,5-cyclooctadiene,ionene,naringenin,methyl ricinole,etc.4 different metabolites were found between one-year pine pollen and two-year pine pollen,namely cumene,1-cyclohexyl-4-piperidone,oleoyl glycine,and 4-octadecylaniline.Mainly concentrated in the pine pollen for the biennium.In the samples compared with fresh pine pollen,6 differential metabolites were found between irradiated sterilized pine pollen and fresh pine pollen,and 12 differences were found between ultra-high temperature instantaneous sterilized pine pollen and fresh pine pollen.9 differential metabolites were found between one-year pine pollen and fresh pine pollen,and 11 differential metabolites were found between two-year pine pollen and fresh pine pollen.In these four comparisons,Fresh pine pollen contains more kaempferol,N-(2-hydroxypropyl)methacrylamide,irradiation sterilization group,ultra-high temperature instant sterilization group,one-year and two-year group pine pollen Contains 2-formylpyridine.8kinds of differential metabolites were found between pine pollen water extract and fresh pine pollen,and 5 of them were found in pine pollen aqueous extracts and 3 in fresh pine pollen.It indicated that the pine pollen water extract had more metabolites,and the fresh pine pollen was extracted directly with water,and its metabolites were less.In the second part,107 metabolites were found in fresh Pinus yunnanensis pollen.The information of molecular ions and fragment ions of these metabolites,including English names,molecular formulas,molecular weights,CAS numbers and so on,was analyzed.These data were input into Thermo Trace Finder LC software,and a database of pine pollen metabolites was preliminarily established.