Research On Traceability Method Of Nanfeng Mandarin And Functional Components In Young Nanfeng Mandarin

Nanfeng Mandarin is a kind of citrus,and a kind of luxury products in Jiangxi Province,its fruit can be used as traditional Chinese medicine raw materials,The young fruit can be used as a traditional Chinese medicine raw materials,The young fruit not only chemical composition is complex,and there are many different kinds of structure,but also contains several classes of different chemical properties of the material,the single detection method is difficult to realize effective analysis on its chemical composition,various analytical techniques combination is imperative.In this experiment,HPLC-ESI-QTOF-MS,SPME-GC-MS,HPLC-DAD and HPLC-QQQ-MS four kinds of analysis and detection technology analysis on the chemical constituents of Nanfeng Mandarin.Carries on the analysis to identify the components of the main substances and provide theoretical basis for the study of Nanfeng Mandarin.Based on the methods of metabonomics,The data will be processed to construct the model of metabolic fingerprint,principal component analysis(PCA)s and partial least square discriminant analysis(PLS-DA)to establish a new method for tracing the origin of Nanfeng Mandarin.Chapter 1: Details the chemical composition,separation methods and the medicinal valu of Nanfeng Mandarin and other citrus at home and abroad in recent years,Citrus fruit is rich in nutrition,not only contains abundant nutrients,such as pectin,oligosaccharides,organic acids and amino acids etc,but also contains a variety of non nutritional physiological active ingredients that have a good effect on physiological and prevent disease to human health,such as limonin,alkaloid,volatile oils,phenols etc.These trace elements make citrus fruits have unique flavor and nutritional value,At the same time,the research status of metabonomics at home and abroad is discussed.At present,there are a lot of applications of metabonomics in Botany,and most of them focus on the study of metabolic profiles and metabolic fingerprints.Chapter 2: Nanfeng Mandarin volatile constituents was extracted by the method of solid phase microextraction,the extraction was isolated and identified by the technique of combination of GC/MS,The experimental method is optimized.73 kinds of volatile substances were preliminary identified,The volatile components of Nanfeng Mandarin include alkenes,alcohols,ketones,aldehydes,esters,phenols and some hydrocarbons.The main volatile components of Nanfeng mandarin fruit is olefin hydrocarbon,accounted for 86.97% of the total volatile components extraction.The highest content was D-limonene,which was the most important component of most citrus fruits.Chapter 3: An ultra-performance liquid chromatography-photodiode array detector-quadrupole time-of-flight mass spectrometry(UPLC-ESI-QTOF-MS)method was established to analyze the chemical component in young Nanfeng Mandarin?Agilent-C18 column(2.1 mm × 100 mm,1.8 ?m,Agilent Technologies,USA)was used with aqueous solution(0.1% formic acid)as mobile phase A and methanol as mobile phase B in gradient elution.Mass spectrometry used ESI ion source,and mass scanning range was m/z100-1000.Through the analysis of the positive and negative ion mass spectra information,and the relevant reports,21 compounds were identified.Analysis of polyphenol content and antioxidant activities of the different origin of young Nanfeng Mandarin,The total phenolics,flavonoids and antioxidant capacity were better.and the total phenolic content and antioxidant capacity were significantly correlated.That indicted the polyphenols may be the main material of antioxidant.Studying on young Nanfeng Mandarin chemical composition and antioxidant activity provide a theoretical basis for the development and utilization of Nanfeng Mandarin,and promote the Nanfeng Mandarin industry development at the same time.Chapter 4: Phenolic acids have many physiological functions and Widely existed in plant tissues.Six phenolic acids in young Nanfeng Mandarin were determined by HPLC-MS in this study,which were: Caffeic acid,p-coumaric acid,ferulic acid,sinapic acid,gallic acid and p-hydroxybenzoic acid.Results show: there were more hydroxycinnamics than hydroxybenzoics both in young Nanfeng Mandarin,and contents of ferulic acid were highest among the six quantified phenolic acids,the second is caffeic acid,The content of soluble phenolic acid is much higher than the content of bound phenolic acid.Chapter 5: Volatile compounds of Nanfeng Mandarin were investigated by solid phase micro extraction(SPME)followed by gas chromatography-mass spectrometry(GC-MS),69 kinds of volatile aroma components were extracted,Principal components analysis was applied to find the main factors affecting the volatile components.The PCA results showed that 6 principal components could reflect most details on Nanfeng Mandarin with a cumulative contribution rate of 84.31%,Most of the information of volatile components in the Nanfeng Mandarin can be reflected.This provides a new idea for the study on volatile components in Nanfeng Mandarin.Chapter 6: Nanfeng Mandarin fingerprint by HPLC was established with comparison of similarity for 18 batches of samples,calibration of the 26 common chromatographic peaks.The 26 common chromatographic peaks are normalized by the SPSS22.0 statistical software program to make principal component analysis.The PCA results showed that 6 principal components could reflect most details on Nanfeng Mandarin with a cumulative contribution rate of 87.50%.The difference appearance of different origin Nanfengmijie is not obvious,but the quality of taste was quite different.The principal component analysis of the common chromatographic peaks make the results of the data analysis is more objective,and greatly reduce the error caused by former subjective judgment.Chapter 7: The Nanfeng Mandarin samples was analyzed and modeled used by significance analysis(ANOVA and Fold Change analysis)?clustering analysis?principal component analysis(PCA)and partial least square discriminant analysis(PLS-DA),Distinguish the different origin of Nanfengmijie as excepted,and 29 potential marker compounds are preliminary acquired.Based on HPLC-ESI-QTOF-MS detecting,and combined with multifarious chemometrics methods to analyze Nanfeng Mandarin,then to set up the fingerprint spectrogram and the model of Nanfeng Mandarin,and to find the characteristic potential marker compounds finally.The method in this study is all-purpose and common approach used in fingerprint spectrogram analysis of the regional characteristic food.