Searching For The Marker Metabolites Of Honey Based On Metabonomics And Exploring Its Application

Honey is rich nutritional value,popular food.Due to the long production cycle and low yields,honey is often in short supply,coupled with its complex composition,make it become a major object of food adulteration.Criminals often adulterate honey through various means to seek benefits.Because the main component of honey is glucose and fructose,the incorporation of cheap way to make syrup adulteration of honey is commonly used means of honey adulteration.Existing method for detecting honey adulteration are: the analysis of stable carbon isotope ratio(SCIRA),thin layer chromatography(TLC),high performance anion exchange pulse current detector chromatography(HPAEC-PAD),gas chromatography-mass spectrometry(GC-MS),high performance liquid chromatography(HPLC),coupled with high performance liquid chromatography isotope mass spectrometer(HPLC-IRMS)and magnetic resonance imaging(NMR)and near infrared spectroscopy(NIR),but these methods all have their own defects,and identifying the adulteration of honey with syrup from C3 plants did not break.This study uses liquid chromatography-mass spectrometry technology to conduct a comprehensive acquisition of metabolites for real honey(chaste honey?acacia honey?date Honey)and adulterated honey,combining with multivariate statistical analysis software to distinguish true honey and honey adulteration,finding maker metabolites of true honey samples,and then using to discriminate true honey and adulterated honey.This study is of great significance to control the quality of honey in the honey market in the future.This experiment is divided into four parts,the four parts are matebolomic study of looking for real honey marker metabolites,distinguishing true honey and syrup used in honey adulteration,distinguishing ripe honey and immature honey,distinguishing ture honey and known inferior honey.The four part of the experiment use ultra high performance liquid chromatography coupled triple quadrupole obitrap high resolution mass spectrometry(UHPLC-Q Exactive Obitrap LC-MS)to obtain the original sample data,the total ion flow chromatographies analyse by Compounds Discoverer software,to extract chromatographic?align peak peak?analysis the unknown biomarker substances,through multivariate statistical analysis,PCA can discriminating comparison samples.Then we find fragment ions of maker matebolites by ms/ms analysis,completing characterization of the marker metabolites preliminary.The first part results showed that the real honey samples of chaste honey?acacia honey?date Honey,could distinguish with various syrup which used in honey adulteration,we found possible maker metabolites of three typies of honey and conducted qualitative analysis.Potential maker matebolimates of chaste honey cantains phenylalanine?leucine?pantothenic acid?p-coumaric acid?cinnamic acid?chrysin?tyrosine,phenylalanine and pantothenic acid are specific marker of chaste honey;Potential maker matebolimates of acacia honey cantains tryptophan?leucine?tyrosine?proline?retinoic acid?fragrant bean acid?abscisic acid?cinnamic acid?chrysin,tryptophan?retinoic acid and abscisic acid are specific markers of acacia honey;Potential maker matebolimates of date honey cantains melatonin ?N-acetylserotonin leucine?tyrosine?proline?p-coumaric acid?cinnamic acid?4-Methoxycinnamic acid ? chrysin,melatonin ? N-acetylserotonin leucine ?4-Methoxycinnamic acid are specific markers of chaste honey.The second part results show: The effection of distinguishing with the three kinds of honey and the syrup used in honey adulteration is obvious,Chaste honey?acacia honey?and date honey adulterated 1% rice syrup?1% corn syrup?1%sugar beet syrup and 1% honey used syrup mixed can really distinguish between honey.In this study,the limit of the matebolomic method of syrup adulteration detection for honey can reach 1%,and with the increase of the proportion of syrup,adulterated honey group and real honey group were getting farther and farther away in the PCA score plot.Mature and immature honey of chaste honey ?acacia honey?date Honey have a obvious separation effect on the first principal componentin in the PCA score plots in the third part.At the end of the test for the identification of true and inferior honey,real acacia honey and known inferior acacia honey can be distinguished by the metabonomics method we used in the research,and real date honey and known inferior date honey have the same result.When we put different varieties of known inferior honey in the comparation with real chaste honey?real acacia honey?real date honey,we found that the three different varieties of known inferior honey are classified as one group,and separated with three varieties of real honey.Combined with the analysis of the results,the effect of identification of honey adulteration is obvious by the metaboomic methods used in this study,and is better than the other methods used in adulterated honey identification.The matebolomic method is not necessary to carry out the qualitative and quantitative determination of one or some target substances,and it is a kind of method of macroscopic non target species differentiation.The method is convenient and fast,and its pretreatment method is simple and easy to operate.Processing data by Compounds Discoverer software,we carried out multivariate statistical analysis to build PCA model,then the real honey and adulterated honey can be seperated.