本文選題：代謝組學(xué) + 產(chǎn)地溯源　； 參考：《中國(guó)農(nóng)業(yè)科學(xué)院》2016年碩士論文

【摘要】：茶葉有著悠久的歷史,是中國(guó)重要的經(jīng)濟(jì)作物。在傳統(tǒng)的名優(yōu)綠茶中,龍井茶,特別是西湖龍井的地位舉足輕重。目前,除西湖龍井外,新昌大佛龍井、嵊州越鄉(xiāng)龍井、磐安生態(tài)龍井、淳安千島湖龍井、蕭山湘湖龍井等龍井茶“區(qū)域子品牌”已呈現(xiàn)快速發(fā)展的勢(shì)頭,形成了茶葉界獨(dú)一無(wú)二的龍井茶“品牌板塊”,也引起了業(yè)界高度關(guān)注。同時(shí),以西湖龍井為例的部分名優(yōu)茶市場(chǎng)依然廣泛存在“以假亂真”的現(xiàn)象,為了給消費(fèi)者一個(gè)明確的判斷標(biāo)準(zhǔn)以及完整的科學(xué)依據(jù),本研究以對(duì)扁形茶產(chǎn)地判定為目的進(jìn)行化學(xué)指紋圖譜的分類(lèi)和判別研究。綠茶是世界主要的健康飲品之一,其內(nèi)在品質(zhì)是評(píng)判綠茶質(zhì)量的主要因子,專(zhuān)家感官審評(píng)是當(dāng)前主要的品質(zhì)等級(jí)評(píng)定方法,然而感官審評(píng)或多或少受審評(píng)者的身體精神狀態(tài)或人為主觀等因素影響,不同的審評(píng)者對(duì)同一茶葉的品質(zhì)評(píng)判可能會(huì)有所差異。在研究了龍井茶原產(chǎn)地判別問(wèn)題的基礎(chǔ)上,建立扁形茶品質(zhì)的預(yù)測(cè)模型和扁形茶等級(jí)的判別模型,對(duì)比影響扁形茶品質(zhì)的特征化合物和影響扁形茶產(chǎn)地的特征化合物,最終確定了不同產(chǎn)地的扁形茶樣品并未與其品質(zhì)有明顯的相關(guān)性。究其根本原因是影響扁形茶品質(zhì)的代謝化合物主要為兒茶素、氨基酸、植物堿、茶黃素以及其他兒茶素類(lèi)聚合物等茶葉中含量較高的化合物,而不同產(chǎn)地間因海拔、緯度與小氣候因素所形成的特征化合物主要為黃酮類(lèi)、多糖類(lèi)與肽類(lèi)等茶葉中含量較低的化合物。本實(shí)驗(yàn)主要研究結(jié)果如下:1對(duì)浙江、四川、貴州、山東四個(gè)產(chǎn)茶省的扁形茶樣品的分析通過(guò)對(duì)浙江、四川、貴州、山東四個(gè)產(chǎn)茶省的扁形茶樣品的代謝指紋圖譜進(jìn)行多元統(tǒng)計(jì)分析,PCA分析結(jié)果R~2X=0.643,Q~2X=0.363,散點(diǎn)圖中各產(chǎn)茶省樣品各自呈現(xiàn)聚合的趨勢(shì);PLS分析結(jié)果R~2X=0.396,R~2Y=0.874,Q~2X=0.898。共鑒定了32種不同產(chǎn)茶省間的特征化合物。包括Cysteinyl-Glycine、3-Nitrotyrosine、Mevalonic acid-5P、N-Carbamoyl-2-amino-2-(4-hydroxyphenyl)acetic acid、3-O-p-Coumaroylquinic acid、Gallic acid4-O-(6-galloylglucoside)等。2對(duì)西湖、錢(qián)塘、越州、縉云四個(gè)產(chǎn)茶區(qū)的扁形茶樣品的分析通過(guò)對(duì)西湖、錢(qián)塘、越州、縉云四個(gè)產(chǎn)茶區(qū)的扁形茶樣品的代謝指紋圖譜進(jìn)行多元統(tǒng)計(jì)分析,PCA分析結(jié)果R~2X=0.730,Q~2X=0.388,散點(diǎn)圖中各產(chǎn)茶省樣品各自呈現(xiàn)聚合的趨勢(shì);PLS分析結(jié)果R~2X=0.437,R~2Y=0.784,Q~2X=0.845。共鑒定了32種不同產(chǎn)茶省間的特征化合物。包括5-Amino-6-(5'-phosphoribitylamino)uracil、4-Methyl-2-phenyl-2-pentenal、Ethyl aconitate、1-(2-Hydroxyphenylamino)-1-deoxy-beta-D-gentiobioside 1,2-carbamate、Imidazoleacetic acid ribotide等。3對(duì)西湖龍井茶保護(hù)區(qū)內(nèi)個(gè)主要產(chǎn)茶村的扁形茶樣品的分析通過(guò)對(duì)西湖龍井茶保護(hù)區(qū)內(nèi)個(gè)主要產(chǎn)茶村的扁形茶樣品的代謝指紋圖譜進(jìn)行多元統(tǒng)計(jì)分析,PCA分析結(jié)果R~2X=0.786,Q~2X=0.464,散點(diǎn)圖中各產(chǎn)茶省樣品各自呈現(xiàn)聚合的趨勢(shì);PLS分析結(jié)果R~2X=0.453,R~2Y=0.848,Q~2X=0.895。共鑒定了44種不同產(chǎn)茶省間的特征化合物。包括4-O-Methylgallic acid、3,4,5-Trimethoxycinnamic acid、Ethyl 2-phenyl-3-furancarboxylate、3-(3,4-Dihydroxyphenyl)lactic acid、Dimethyl tetrasulfide等。4影響扁形茶品質(zhì)的特征化合物的鑒定通過(guò)三個(gè)等級(jí)來(lái)鑒定了水提取結(jié)合UPLC-Q-TOF/MS測(cè)到的1568種代謝化合物中的60種化合物。通過(guò)t檢驗(yàn)得到其中23種化合物在A、B、C三個(gè)區(qū)域間具有顯著性差異且三個(gè)區(qū)間的樣品審評(píng)得分遞減,其中L-Arginine與L-Glutamine等氨基酸類(lèi)化合物在區(qū)域A與區(qū)域C之間具有顯著性差異,得出該類(lèi)化合物與扁形茶品質(zhì)成正相關(guān)。與氨基酸類(lèi)化合物相反,Serinyl-Serine等三種多肽類(lèi)化合物在A區(qū)域最低C區(qū)域最高,表現(xiàn)出了與品質(zhì)的負(fù)相關(guān)性。在兒茶素類(lèi)化合物中Epigallocatechin表現(xiàn)出了與綠茶品質(zhì)的負(fù)相關(guān)性,Catechin僅在B區(qū)域的含量顯著低于C區(qū)域,其余Gallocatechin,Gallocatechin gallate,Catechin gallate,Epigallocatechin gallate等四種化合物都在不同程度上表現(xiàn)出了與品質(zhì)成正相關(guān)性。包括Theaflavin與Catechin-(4beta-8)-Epigallocatechin在內(nèi)的茶黃素類(lèi)物質(zhì)和黃烷醇類(lèi)化合物的聚合物均表現(xiàn)出了與綠茶品質(zhì)的負(fù)相關(guān)。因此,可初步認(rèn)為,這些特征化合物可用來(lái)進(jìn)行扁形綠茶品質(zhì)的預(yù)測(cè)。5甲醇提取條件與水提取條件下扁形茶品質(zhì)與等級(jí)預(yù)測(cè)模型的對(duì)比分析在高通量色譜質(zhì)譜技術(shù)中,水提取條件下得分預(yù)測(cè)模型(PLS回歸模型)的RMSEP值小于甲醇提取條件,而R~2、Q~2值大于甲醇提取條件。在等級(jí)預(yù)測(cè)模型中,決策樹(shù)、神經(jīng)網(wǎng)絡(luò)和貝葉斯網(wǎng)絡(luò)等模型的準(zhǔn)確度都在在水提取條件下達(dá)到90%以上,而預(yù)測(cè)度均達(dá)到80%以上,遠(yuǎn)高于甲醇提取條件下準(zhǔn)確度72%的均值和預(yù)測(cè)度62%的均值。這表明,相對(duì)傳統(tǒng)的甲醇提取方法,在UPLC-Q-TOF/MS平臺(tái)中水提取方法更適合進(jìn)行綠茶審評(píng)得分或等級(jí)的預(yù)測(cè)。
[Abstract]:Tea has a long history and is an important economic crop in China. In the traditional famous green tea, Longjing tea, especially West Lake Longjing, is very important. At present, besides West Lake Longjing, Xinchang Grand Buddha Longjing, Shengzhou Yue Township Longjing, Panan ecological Longjing, Chunan Qiandao Lake Longjing, etc. The momentum of rapid development has formed the unique "brand plate" of Longjing tea industry in the tea industry, which has also aroused great concern in the industry. At the same time, some famous tea markets in Longjing, West Lake, still exist widely in the "false and true" phenomenon, in order to give consumers a clear standard of judgment and a complete scientific basis, the research is a complete scientific basis. The purpose of the study is to classify and discriminate the chemical fingerprint with the purpose of determining the producing area of the flat tea. Green tea is one of the main health drinks in the world. Its intrinsic quality is the main factor to judge the quality of green tea. The expert sensory evaluation is the main quality evaluation method at present, but the sensory evaluation is more or less subject to the body essence of the reviewers. Different judges may vary the quality of the same tea. Based on the study of the identification of Longjing tea origin, the prediction model of the quality of the flat tea and the discriminant model of the level of the flat tea are established, and the characteristic compounds and the flat tea which affect the quality of the flat tea are compared. The fundamental reason is that the metabolic compounds that affect the quality of the flat tea are mainly catechins, amino acids, alkaloids, theaflavins, and other catechin polymers, and the different compounds in the tea. The main research results are as follows: 1 the analysis of flat tea samples from four tea producing provinces in Zhejiang, Sichuan, Guizhou and Shandong was analyzed through the analysis of four tea producing tea in Zhejiang, Sichuan, Guizhou and Shandong. The metabolic fingerprint of the sample of the flat tea of the province was analyzed by multivariate statistical analysis. The results of PCA analysis were R~2X=0.643, Q~2X=0.363, and each tea producing province in the scatter plot showed the trend of polymerization. The results of PLS analysis, R~2X=0.396, R~2Y=0.874, Q~2X=0.898., identified the characteristic compounds of 32 different tea producing provinces, including Cysteinyl-Glycine, 3-Nitrotyro Sine, Mevalonic acid-5P, N-Carbamoyl-2-amino-2- (4-hydroxyphenyl) acetic acid, 3-O-p-Coumaroylquinic acid, Gallic acid4-O- (6-galloylglucoside), etc. Analysis of the flat tea samples from four tea producing areas in West Lake, Qian Tang, Yue Zhou and Jinyun through the metabolic fingerprint of the flat tea samples from four tea producing areas in West Lake, Qian Tang, Yue Zhou and Jinyun. Multivariate statistical analysis of the atlas, PCA analysis results R~2X=0.730, Q~2X=0.388, each tea producing province sample in the scatter plot showed the trend of polymerization; PLS analysis results R~2X=0.437, R~2Y=0.784, Q~2X=0.845. have identified 32 different tea producing provinces, including 5-Amino-6- (5'-phosphoribitylamino) uracil, 4-Methyl-2-phenyl-2-pent Enal, Ethyl aconitate, 1- (2-Hydroxyphenylamino) -1-deoxy-beta-D-gentiobioside 1,2-carbamate, Imidazoleacetic acid Ribotide and so on, the analysis of the flat tea samples from the main tea producing village in West Lake Longjing tea protection area was carried out by the metabolic fingerprint of the flat tea samples from the main tea producing village in the West Lake Longjing tea reserve. Multivariate statistical analysis, PCA analysis results R~2X=0.786, Q~2X=0.464, the scatter plot of the tea production samples each showed the trend of polymerization; PLS analysis results R~2X=0.453, R~2Y=0.848, Q~2X=0.895. identified 44 different tea producing provinces, including 4-O-Methylgallic acid, 3,4,5-Trimethoxycinnamic acid, Ethyl Boxylate, 3- (3,4-Dihydroxyphenyl) lactic acid, Dimethyl tetrasulfide and other.4 that affect the quality of flat tea, identified 60 compounds of the 1568 metabolic compounds detected by water extraction combined with UPLC-Q-TOF/MS by three grades. By t test, 23 of these compounds were found in A, B, and three regions. The score of sample review in the three interval is decreasing, in which the amino acid compounds such as L-Arginine and L-Glutamine have a significant difference between the regional A and the regional C. It is concluded that the compounds are positively related to the quality of the flat tea. Contrary to the amino acids, the three kinds of polypeptide compounds such as Serinyl-Serine are the lowest in the A region C. In the catechin compounds, Epigallocatechin showed a negative correlation with the quality of green tea, and the content of Catechin only in the B region was significantly lower than that in the C region. The remaining Gallocatechin, Gallocatechin gallate, Catechin gallate, Epigallocatechin gallate and other four compounds were different. The quality is positively correlated with the quality. The theaflavins and the flavanin compounds, including Theaflavin and Catechin- (4beta-8) -Epigallocatechin, are negatively correlated with the quality of green tea. Therefore, it is preliminarily believed that these compounds can be used to predict the quality of flat green tea,.5 a Comparison and analysis of the quality and grade model of flat tea under the conditions of alcohol extraction and water extraction, in high flux chromatography-mass spectrometry, the RMSEP value of the prediction model (PLS regression model) under the water extraction condition is less than the methanol extraction condition, while the R~2, Q~2 value is greater than the methanol extraction strip. The accuracy of the Bias network model is more than 90% under the water extraction condition, and the prediction degree is above 80%, which is far higher than the mean value of the accuracy of 72% and the mean of 62% in the methanol extraction condition. This shows that the water extraction method in the UPLC-Q-TOF/MS platform is more suitable for green tea review than the traditional methanol extraction method. A prediction of a score or grade.
【學(xué)位授予單位】：中國(guó)農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S571.1

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