气质联用在不同茶叶品质鉴定中的应用
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
茶树为多年生常绿叶用作物,茶叶是茶树鲜叶经过工艺处理后制作而成。乌龙茶学名青茶,为我国六大茶类之一,主产于我国的福建、台湾和广东等省。乌龙茶属于半发酵茶,同一茶树品种因地域不同其品质也不一样,其中铁观音品种中在福建安溪县范围内的品质最佳。本论文基于气相色谱质谱联用(GC-MS)的非靶标代谢轮廓分析方法,开展安溪铁观音代谢组学研究,旨在从代谢组学的角度研究安溪县铁观音的品质与真伪。
     建立了用于茶叶研究的衍生化GC-MS的方法,对预处理条件进行了全面考察和优化,确定了样品预处理方法,用70%的甲醇水溶液作为最佳提取溶剂。同时对方法进行了表征(标准曲线、精密度实验、回收率等),该方法满足分析的需要。采用该方法对茶叶进行了分析,共鉴定了茶叶中69种代谢物,包括7种糖,14种氨基酸,20种有机酸。
     采用本论文建立的衍生化GC/MS方法,对安溪县境内8个乡镇种植的铁观音及其安溪县种植的三个乌龙茶品种(黄金桂、本山和毛蟹)进行代谢组学分析研究,PCA结果表明安溪县境内不同乡镇的铁观音表型有明显的差异,但相邻乡镇的茶叶代谢表型相似,为辨别安溪铁观音真伪奠定了基础,在国内尚未见过报道。
     用衍生化GC/MS方法还对绿茶、白茶、乌龙茶(铁观音和水仙)和红茶进行代谢组学研究,用主成分分析对数据进行处理,PCA结果表明不同发酵程度的四种茶叶代谢表型差异显著,同为乌龙茶的铁观音和水仙在PCA得分图上距离较近但也能清楚分类,发酵茶红茶与其它品种茶叶的差异最大。
Tea tree is an evergreen shrub whose leaves are used to produce tea through processing. Tea quality varied widely according to type of tea and geographical origin et al. Semi-oxidised oolong tea is called blue-green tea, which is one of the six main types of Chinese teas. Major Oolong tea growing regions are Fujian, Taiwan, and Guangdong provinces. Anxi Tieguanyin is one of the best brands of the Oolong teas. In this thesis, Anxi Tieguanyin teas were investigate based on GC-MS non-targeted profiling method. The main results were as follows:
     A GC-MS metabolomics method for tea analysis was established. Sample pretreatment method was developed. Extraction solvent and derivatization conditions were investigated and optimized. The optimal extraction solvent was 70% methanol solution. Validation of the method was carried out. Repeatability, recovery and linearity results showed that the developed method could meet the analytical requirement. Using the developed method, a typical Tieguanyin was analyzed. A total of 69 metabolites were tentatively identified, including 7 sugars,14 amino acids,20 organic acids.
     Then, Tieguanyin and three main Oolong tea varieties (Benshan, Mao Xie and Huang Jin Gui) originated from eight towns within Anxi country were analyzed using the developed method. The PCA score plot showed that the phenotype of Oolong tea from different towns varied widely. Tieguanyin from geographically adjacent regions were clustered.
     Finally, four main types of tea including green tea, white tea, Oolong tea (Tieguanyin and Shui Xian) and the black tea were investigated. The PCA score plot indicated obvious discrimination of teas according to their degrees of fermentation. Tieguanyin and Shui Xian belong to the Oolong tea, and they are clustered nearby on PCA score plot. The phenotype of black tea have the biggest difference from other types of tea.
引文
[1]Nicholson J.K, Lindon J.C, Holmes E.'Metabonomics':understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J]. Xenobiotica,1999,29(11):1181~1189.
    [2]Kanani H., Chrysanthopoulos P.K., Klapa M.I., Standardizing GC-MS metabolomics [J]. J Chromatogr B,2008,871:191~201.
    [3]Fiehn O., Extending the breadth of metabolite profiling by gas chromatography coupled to mass spectrometry [J]. Trends Analyt Chem,2008,27:261~269.
    [4]Wang T.J., Larson M.G., Vasan R.S., Cheng S., et al., Metabolite profiles and the risk of developing diabetes[J]. Nat Med,2011,17:448~453.
    [5]Chen, C.; Gonzalez, F.J.; Idle, J.R., LC-MS-based metablmics in drug metabolism J]. Drug Metabolism Review 2007,39 (2-3),581~597.
    [6]Shulaev, V.; Cortes, D.;Mittler,R., Metabolomics for plant stress response [J]. Physiologia Plantrum,2008,132(2),199~208.
    [7]Giri S, Idle J R, ChenC, et al. A metabolomic approach to the metabolism of the areca Nut alkaloids arecoline and arecaidine in the mouse[J]. Chem Res Toxicol,2006,19(6): 818~827.
    [8]Antonio Moreda-Pineiro, Andrew Fisher, Steve J. Hill.The classification of tea according to region of origin using pattern recognition techniques and trace metal data[J]. Journal of Food Composition and Analysis,2003,16:195~211.
    [9]LindonJC, HolmesE, NicholsonJK. So what is the deal with metabonomics?[J]. Anal Chem,2003,75(17):384A~391A.
    [10]Fiehn O., Kopka J., Dormann P., Altmann T., et al., Metabolite profiling for plant functional genomics [J]. Nat Biotechnol,2000,18:1157~1161.
    [11]许国旺,路鑫,杨胜利。代谢组学研究进展[J].中国医学科学院学报,2007,29(6):701~711.
    [12]Nicholson J.K., Wilson I.D., Opinion:understanding 'global' systems biology: metabonomics and the continuum of metabolism [J].Nat Rev Drug Discov,2003,2:668~676.
    [13]彭超,黄和,肖爱华等,代谢组学分析技术平台及方法研究进展[J].食品安全与检测,2008,9:220~223.
    [14]Fiehn O, Combining genomics, metabolome analysis and biochemical modeling to understand Metabolic networks[J]. Comparative and Functional Genomics,2001,2:155~168.
    [15]Fiehn O, Kopka J, Dormann P, et al. Nat. Biotechnol.,2000,18(11):1157.
    [16]夏建飞,梁琼麟,胡坪等。代谢组学研究策略与方法的新进展[J],分析化学(FENXI HUAXUE)评述与进展,2009,37(1):136~143.
    [17]刘祥东,罗国安,王义明,代谢物组学应用的领域之二——植物代谢物组学[J],中成药,2006,28(10)1515.
    [18]李忠,杨洪明,徐世涛等,基于质谱的代谢组学方法在烟草化学中的应用研究,2007,z1期,分析测试学报,294-295.
    [19]李忠,杨洪明,徐世涛等。基于质谱的代谢组学方法在烟草化学中的应用研究[J].分析测试学报,2007,26卷增刊:294~296.
    [20]杨军,宋硕,Jose Castro Per等,代谢组学及其应用[J].生物工程学报,2006,21(1):1-5.
    [21]宛晓春,茶叶生物化学,第三版[M]。中国农业出版社,2003,39-47.
    [22]Wipawee Pongsuwan, Eiichiro Fukusaki, Takeshi Bamba, et al. Prediction of Japanese Green Tea Ranking by Gas Chromatography/Mass Spectrometry-Based Hydrophilic Metabolite Fingerprinting [J]. Agric. Food Chem.,2007,55:231~236.
    [23]Kanokwan Jumtee, Takeshi Bamba, Eiichiro Fukusaki. Fast GC-FID based metabolic fingerprinting of Japanese green tea leaf for its quality ranking prediction [J]. Sep. Sci.,2009, 32:2296~2304.
    [24]TATSUHIKO IKEDA, SHIGEHIKO KANAYA, TSUTOMU YONETANI, et al. Prediction of Japanese Green Tea Ranking by Fourier Transform Near-Infrared Reflectance Spectroscopy[J]. Agric. Food Chem.2007,55:9908~9912.
    [25]Wipawee Pongsuwan, Takeshi Bamba, Kazuo Harada, et al. High-Throughput Technique for Comprehensive Analysis of Japanese Green Tea Quality Assessment Using Ultra-performance Liquid Chromatography with Time-of-Flight Mass Spectrometry (UPLC/TOFMS)[J]. Agric.Food Chem.,2008,56:10705~10708.
    [26]王丽鸳,成浩,周健,茶树DNA分子标记及基因工程研究进展[J],茶叶科学,2004,24(1),12-17.
    [27]Pedro L, Fernandez-Caceres, Maria J etal. Differentiation of Tea (Camellia sinensis) Varieties and Their Geographical Origin According to their Metal Content [J], J. Agric. Food Chem.2001,49,4775~4779.
    [28]Guoxiang Xie, Mao Ye, Yungang Wang, etal. Characterization of Pu-erh Tea Using Chemical and Metabolic Profiling Approaches [J]. Agric. Food Chem.,2009,57:3046~ 3054.
    [29]DunnWB, BaileyNJ, JohnsonHE. Measuringthemetabolome:current analytical technologies[J]. Analyst,2005,130(5):606~625.
    [30]唐惠儒,王玉兰。代谢组学:一个迅速发展的新兴学科[J].生物化学与生物物理进展,2006,33(5):401-417.
    [31]Carsten Denkert, Jan Budczies, Tobias Kind.Mass Spectrometry-Based Metabolic Profiling Reveals Different Metabolite Patterns in Invasive Ovarian Carcinomas and Ovarian Borderline Tumors[J], Cancer Res,2006,66(22):10795~10804.
    [32]黄瑛。代谢组学在药物毒理学中的应用[J],药学实践杂志,2009,27(3):165-167.
    [33]许国旺,杨军。代谢组学及其研究进展[J].色谱,2003,21(4):316~320.
    [34]蔡爽,孙博,李发美。代谢组学及其在药学中的应用[J].沈阳药科大学学报,2007,24(7):445-450.
    [35]尹恒,李曙光,白雪芳等。植物代谢组学的研究方法及其应用[J].植物学通报,2005,22(5):532~540.
    [36]黄强,尹沛源,路鑫等。色谱-质谱联用技术在代谢组学中的应用[J],色谱(Chinese Journal of Chromatography,2009,27(5):566~572.
    [37]施钰琳,王小琴。代谢组学及其应用研究新进展[J],医学综述,2007,13(20):1543~1545.
    [38]Schaub J,Schiesling C, Reuss M. Integrated sampling procedure for metabolome analysis[J], Biotechno Progr,2006,22(5):1434~1442.
    [39]Dettmer K, Aronov PA, Hammock BD. Mass spectrometry—based metabolomics[J]. Mass Spectrometry Reviews,2007,26(1):51~78.
    [40]Goodacre R, Vaidyanathan S, Dunn W B, etal. Metabolomics by numbers:acquiring and understanding global metabolite data[J]. Trends Biotechnol,2004,22(5):245~252.
    [41]Villas-Boas SG, Mas S, Akesson M, et al. Mass spectrometry in metabolome analysis [J]. Mass Spectrometry Reviews,2005,24(5):613~646.
    [42]Brown M, Dunn W B, EHis DI, etal. A metabolome pipeline:from concept to data to knowledge[J], Metabolomics,2005,1(1):39~51.
    [43]Yang J, Xu G, Zheng Y, etal. Strategy for metabonomics research based on high-performance liquid chromatography and liquid chromatography coupled with tandem mass spectrometry[J], J Chromatogr A.2005。1084(1-2):214~221.
    [44]郭宾,戴仁科。代谢组学及其研究策略和分析方法进展[J].中国检验卫生杂志,2007,17(3):554~563.
    [45]淡漠,高先富,谢国祥。代谢组学在植物代谢研究中的应用[J],中国中药杂志,2007,32(22):2337-2341.
    [46]董登峰。代谢物组学方法及其在植物学研究中的应用[J],广西植物,2007,27(5):765~769.
    [47]卢红梅,梁逸曾。代谢组学分析技术及数据处理技术[J],分析测试学报,2008,27(3):325~332.
    [48]焦旭雯,赵树进。药用植物代谢组学的研究进展[J],广东药学院学报,2007,23(2):228~231.
    [49]焦玉红,刘志友,王发善。代谢组学在医药领域的应用和前景展望[J],现代企业文化,2009,23:9-9.
    [50]Cloarec O, Campbell A, Tseng L H,etal. Virtual Chromatographic Resolution Enhancement In Cryoflow LC-NMR Experiments via Statistical Total Correlation Spectroscopy[J],2007,79(9):3304~3311.
    [51]Ong E S, Chor C F, Zou L, etal. A multi-analytical approach for metabolomic profiling of zebrafish (Danio rerio) livers[J], Mol Biosyst,2009,5(3):288~298.
    [52]Lin Y Q, Schiavo Susan, Orjala J, etal. Microscale LC-MS-NMR Platform Applied to the Identification of Active Cyanobacterial Metabolites[J], Anal Chem.,2008,80(25): 8045~8054.
    [53]J. William Allwood, David I. Ellis and Royston Goodacre. Metabolomic technologies and their application to the study of plants and plant-host interactions [J]. Physiologia Plantarum,2008,132:117~135.
    [54]刘昌孝。代谢组学与中药现代研究[J].河南大学医学学报(医学版),2006,25(3):1-7.
    [55]S.Kravic, Z.Suturovic, J.Svarc-Gajic, Z. Stojanovic etal. Determination of trans fatty acids in foodstuffs by gas chromatograpy-mass spectrometry after simultaneous microwave-assisted extraction-esterification.[J] Journal of the Serbian Chemical Societ.2010, 75(6),803~812.
    [56]X. F. Yue, Y. N. Zhang, Z. Q. Zhang. Free fatty acids profile analysis of alcohol extract of Aconitum taipeicum Hang-Mazz.with gas chromatograpy-mass spectrometry. [J] Analytical Methods,2010,2(6),668~672.
    [57]M. Petrovic, N. Kezic, V. Bolanca. Optimization of the GC method for routine analysis of the fatty acid profilein several food samples. [J] Food Chemistry,2010,122(1),285~291.
    [58]Z. Y. Yang, B. P. Hollebone, Z. D. Wang, etal. Determination of polar impurities in biodiesels using solid-phase extraction and gas chromatograpy-mass spectrometry. [J] J Sep. Sci.,2011,34(4),409~421.
    [59]Little,J. L., Artifracts in trimethylsilyl derivatization reactions and ways to avoid them[J], Journal of Chromatogram A 1999,844(1-2),1~20.
    [60]Huskova, R.; Bartak, P.; Cap, L. etal. Analytical derivatization as a tool for determination of orotic acid [J], Journal of Chromatogram B-Analytical Technologies in the Biomedical and Life Sciences.2004,799,(2),303~309.
    [61]Proestos, C.;Sereli, D.; Komaitis, M., Determination of phenolic compounds in aromatic plants by RP-HPLC and GC-MS [J], Food Chemistry 2006,95(1),44~52.
    [62]Zs. F"uzfai, I. Boldizs'ar, I. Moln'ar-Perl, Characteristic fragmentation patterns of the trimethylsilyl and trimethylsilyl-oxime derivatives of various saccharides as obtained by gas chromatography coupled to ion-trap mass spectrometry. [J] Journal of Chromatography A, 2008,1177,183~189.
    [63]Jan Szopa, Grzegorz Wilczyn'ski, Oliver Fiehn etal.Identification and quantification of catecholamines in potato plants (Solanum tuberosum) by GC-MS [J]. Phytochemistry,2001,58,315~320.
    [64]Oliver Fiehn. Metabolic networks of Cucurbita maxima phloem [J]. Phytochemistry, 2003,62,875~886
    [65]Tom Shepherd, Gary Dobson, Susan R. Verrall,etal. Potato metabolomics by GC-MS: what are the limiting factors? [J]. Metabolomics,3(4),475~488.
    [66]Zs. F"uzfai, I. Boldizs'ar, I. Moln'ar-Perl. Characteristic fragmentation patterns of the trimethylsilyl and trimethylsilyl-oxime derivatives of various saccharides as obtained by gas chromatography coupled to ion-trap mass spectrometry [J]. Journal of Chromatography A, 2008,(1177),183~189.
    [67]Horvath,K., MolnarPerl, Simultaneous quantitation of mono-,di-,and trisaccharides by GC-MS of theirTMS ether oxime derivatives. Chromatographia, [J].1997,45,328~335.
    [68]Sangster T., Major H., Plumb R., etal.A pragmatic and readily implemented quality control strategy for HPLC-MS and GC-MS based metabonomic analysis [J]. Analyst,2006, 131,1075~1078.
    [69]Zelena E., Dunn W.B., Broadhurst D.,etal. Development of a robust and repeatable UPLC for the long-term metabolomic study of human serum[J]. Anal Chem.,2009,81, 1357~1364.
    [70]H. M. Lu, W. B. Dunn, H. L. Shen etal. Comparative evaluation of software for deconvolution of matabolomics data based on GC-TOF-MS [J]. Trac-Trend Anal Chem., 2008,27(3),215~227.
    [71]H. R. Norli, A. Christiansen, B. Holen. Independent evaluation of a commercial deconvolution reporting software for gas chromatograpy-mass spectrometry analysis of pesticide residues in fruits and vegetables [J]. Journal of Chromatogram A,2010,1217(13), 2056~2064.
    [72]Smith C.A., Want E.J., O'Maille G., Abagyan R., etal., XCMS:Processing mass spectrometry data for metabolite profiling using Nonlinear peak alignment, matching, and identification [J]. Anal Chem,2006,78:779~787.
    [73]P.Y. Yin, P.Mohemaiti, J.Chen, etal. Serum metabolic profiling of abnormal savda by lipid chromatograpy-mass spectrometry. J Chromatogr B,2008,871(2),322~327.

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700