摘要
产地固有的气候和土壤条件在很大程度上影响着葡萄酒的质量,使它们产生成分差异。对于不同产地的同品种葡萄酒来说,遗传没有影响葡萄的化学成分,且葡萄化学成分的变化也没有规律。因此想要识别不同品种的葡萄酒,只能通过综合考虑建立葡萄酒产地指纹图谱以及葡萄酿酒品种识别模式及数据库。由于葡萄酒中存在许多的花色素及其花色素衍生物,这些物质与葡萄酒的颜色密切相关,所以对花色素及其花色素衍生物的鉴定一直是葡萄酒的分析重点。将指纹图谱技术引入葡萄酒酿造品种鉴别研究,旨在建立一种鉴别葡萄酒酿造品种的简便、可操作性强的客观方法,从而实现指纹图谱方法在葡萄酒酿造品种上的应用。
在前人的研究基础上,本实验对利用指纹图谱技术鉴别葡萄酒酿造品种进行了研究,实验以高效液相色谱法为基础,通过分析大量样品的花色素及其花色素衍生物,筛选出液相色谱测定葡萄酒酿造品种花色素苷的方法和条件,通过研究不同品种葡萄的花色素在种类及含量上的区别,进而建立鉴别葡萄酒酿造品种标准HPLC指纹图谱,并结合数学统计方法建立酿造品种的识别模式。
研究结果:
(1)建立了高效液相色谱测定花色素苷的方法与条件:采用梯度洗脱,流速1.000 mL/min。流动相梯度如下,使用A、B两种流动相,其中A液为乙腈–磷酸缓冲液(v/v=5/95,);B液为乙腈–磷酸缓冲液(v/v=50/50)。进样体积为20μL。检测波长518 nm,柱温50℃。
(2)建立了不同酿造品种葡萄酒HPLC花色素指纹图谱。建立了不同品种葡萄酒HPLC指纹图谱和适合的识别模式,
(3)葡萄酒不同酿造品种的花色素反映了不同酿造品种的遗传信息,成功利用花色素HPLC指纹图谱对贺兰山东麓的几种红色酿造品种葡萄酒进行区分。
(4)应用HPLC花色素指纹图谱及识别模式,准确的对葡萄酒酿造品种、掺假(包括掺水、添加天然色素)葡萄酒进行鉴别。
To a large extend, nature climate and soil conditions affected the quality of wine and lead to different composition in it. But to the different origin wine of the same variety, inheritance do not influence the chemical composition and there is no changing discipline in the chemical composition. Hence, in order to identify the different variety wine, only through the establishment of origin fingerprint of wine, the identification mode and database. There are many anthocyanin and its ramification in wine, and it is closely related to wine color, so the identification to the anthocyanin and its ramification is a focusing part of the wine analysis. In order to establish a simple, efficient and objective method to identify wine grape variety,Fingerprint technology was introduced to the wine making industry.
Based on the previous study, the experiment is to identify wine making varieties by using fingerprint technology. The experiment is based on HPLC, analyzed a large number of samples’anthocyanin and found out the method and condition of HPLC to determine the anthocyanins in different variety’s wine. Based on the research of the distinction of anthocyanin’s variety and content in different wine, to establish the different wine variety’s HPLC fingerprint, and the identification mode of wine making variety also can be established by mathematical statistical method.
The main results were as follows:
(1)Established the method and condition of HPLC to determine the anthocyanins : the flow rate, 1.000 mL/min. The flowing gradient is as follows, uses A and B two kinds flows, solvent A is acetonitrile-phosphoric acid buffer solution (v/v=5/95,); solvent B is acetonitrile -phosphoric acid buffer solution (v/v=50/50). the inject volume, 20μL.; the detecting wavelength, 518 nm;The temperature is 50℃。
(2)Established the HPLC fingerprint of anthocyanins in different variety wine’s; established the HPLC fingerprint and the identification mode of wine making variety.
(3)The anthocyanins of different wine making variety reflect the genetic information of different variety, the different red varietl wine, produced in the east of Helan mountain, be successfully differentiated by the HPLC fingerprint of anthocyanins.
(4)Applied the HPLC fingerprint of anthocyanins and identification mode to evaluate wine quality. Using of HPLC fingerprint can accurately identify the varieties of wine brewing, adulterated wine (including water mixing, add the natural pigment).
引文
[1]李华.葡萄酒的原产地域命名与感官评价[A].李华.第二届国际葡萄与葡萄酒学术研讨会论文集[C].西安:陕西人民出版社.2001,29-34.
[2]耿兆林.为我国葡萄酒行业的持续、健康发展而努力[J].中外葡萄与葡萄酒.2004.4:3-6.
[3] GB15038-2006,《葡萄酒果酒通用试验方法》[S].
[4] GB2758-81《发酵酒卫生标准》[S].
[5] GB2760-96《食品添加剂使用卫生标准》)[S].
[6]马佩选.中国葡萄酒质量体系的建立与展望[J].第二届国际葡萄与葡萄酒学术研讨会论文集,2001:18-22.
[7]李华.中国的原产地域产品与葡萄酒的地理标志及保护[A].李华.第二届国际葡萄与葡萄酒学术研讨会论文集[C].西安:陕西人民出版社.2001,1-5.
[8] Antonieta E M. The influence of climatic variability on the quality of wine[J]. International Journal of Biometeorology, 2001, 45:13–21.
[9]谢培山.色谱指纹图谱分析是中草药质量控制的可行策略[J].中药新药与临床药理,2001,24(3):141-154.
[10]谢培山.中药色谱指纹图谱质量控制模式的研究和应用[J].世界科学技术—中药现代化.2001,3(6):28-32.
[11]任德权.中药质量控制的里程碑-中药指纹图谱[J].中成药, 2001, 23(1):1-2.
[12]罗国安,王义明.中药指纹图谱的分类和发展[J].中国新药杂志, 2002,11(1):46-52.
[13]陈闽军,瞿海斌,程翼宇.一类用化学指纹特征鉴别中药材真伪的方法[J].高等学校化学学报, 2003,24(12):2181-2185.
[14]钱浩泉,李彩君,谢培山.高良姜及其近缘植物挥发油的气相色谱及其指纹图谱研究[J].中药新药与临床药理,2001,12(3):179-182.
[15]谢培山.中药色谱指纹图谱鉴别的概念、属性、技术与应用[J].中国中药杂志, 2001,26(10):653-658.
[16]陈闽军,程翼宇.中药色谱指纹图谱相似性计算方法的研究[J].中成药,2002, 24( 12):905-909.
[17]崔秀明,董婷霞.三七及其混淆品的HPLC指纹图谱鉴定[J].中草药, 2002,33(10):941-944.
[18]刘慧琼,郭书好.现代仪器与中草药指纹图谱[J].中医药学报, 2002,30 (2):35-38.
[19]陈勇,杨文清,蔡敏等.板蓝根药材的HPLC特征指纹图谱研究[J].中药研究,2003,31(3):18-20.
[20]王继国,雍克岚,陈旭等.中药血竭的化学模式识别研究[J].上海大学学报(自然科学版), 2004,10(1):75-78.
[21]孟庆华,刘永锁,王健松等.色谱指纹图谱相似度的新算法及其应用[J].中成药, 2003,25(1):4-8.
[22]徐永群,黄昊,周群等.红外指纹图谱和聚类分析法在赤芍产域分类鉴别中的应用[J].分析化学, 2003,31(1):5-9.
[23]周群,孙素琴,梁曦云.枸杞产地的红外指纹图谱与聚类分析法研究[J].光谱学与光谱分析[J], 2003,23(3):509-511.
[24]沈育杰,赵淑兰,李晓红等.山葡萄种质资源主要经济性状鉴定与评价[J].河北林业科技,2004,5:43-44.
[25]李华.葡萄酒品尝学[M].西安,陕西人民出版社.1994,28-30.
[26]王华,丁刚.葡萄酒中花色素苷研究现状[J].中外葡萄与葡萄酒,2002,2:25-29.
[27]李华.葡萄酒香味成分与感官质量.葡萄栽培与酿酒[J],1990,(3):35-47.
[28] Revilla I, Perez-Magarino S, Gonzalez-SanJose M L, Beltran S. Identification of anthocyanin derivatives in grape skin extracts and red wines by liquid chromatography with diode array and mass spectrometric detection[J]. Journal of Chromatography A, 1999, 847:83-90.
[29] Csomós E, Hèberger K, and Simon-Sarkadi L. Principal Component Analysis of Biogenic Amines and Polyphenols in Hungarian Wines[J]. Journal of Agricultural and Food Chemistry, 2002, 50: 3768-3774.
[30] Bednar P, Tomassi A V, and Presutti C. Separation of Structurally Related Anthocyanins by MEKC[J]. Chromatographia, 2003, 58: 283-287.
[31] Edelmann A, Diewok J, Schuster K C, and Bernhard L. Rapid Method for the Discrimination of Red Wine Cultivars Based on Mid-Infrared Spectroscopy of Phenolic Wine Extracts[J]. Journal of Agricultural and Food Chemistry, 2001, 49:1139-1145.
[32] Revilla E, García-Beneytez E, Cabell F, Mart?n-Ortega G, Jose-Mar?a R. Value of high-performance liquid chromatographic analysis of anthocyanins in the differentiation of red grape cultivars and red wines made from them[J]. Journal of Chromatography A, 2001, 915: 53–60.
[33] Yokotsuka K and Nishino N. Extraction of anthocyanins from Muscat bailey A grape skin[J]. Journal of fermentation and bioengineering, 1990,69(6):328-334.
[34] Marx R, Holbach B, and Otteneder H. Determination of nine characteristic anthocyanins in wine by HPLC[J]. Office of International Vigneand Vin, 2000.
[35] Rosillo L, Salinas M R, Garijo J, and Gonzalo L A. Study of volatiles in grapes by dynamic headspace analysis Application to the differentiation of some Vitis vinifera varieties[J]. Journal of Chromatography A, 1999, 847:155-159.
[36] Ramos R, Andrade P B, Seabra R M, Pereira C, Ferreira M A, Faia M A. A preliminary study of non-coloured phenolics in wines of varietal white grapes (coódega, gouveio and malvasia fina) [J]. Food Chemistry, 1999, 67:39-44.
[37] Sakkiadi A V, Stavrakakis M N, and Haroutounian S A. Direct HPLC Assay of Five Biologically Interesting Phenolic Antioxidants in Varietal Greek Red Wines[J]. Lebensm.-Wiss. u.-Technol., 2001, 34:410-413.
[38] Pozo-Bayón A M, Hernández M T, and Martín-álvarez P J. Study of Low Molecular Weight Phenolic Compounds during the Aging of Sparkling Wines Manufactured with Red and White Grape Varieties[J]. Journal of Agricultural and Food Chemistry, 2003, 51: 2089-2095.
[39] González-Neves G, Charamelo D, Balado J, Barreiro L, Bochicchio R, Gatto G, Gil G , Tessore A, Carbonneau A, Moutounet M. Phenolic potential of Tannat, Cabernet Sauvignon and Merlot grapes and their correspondence with wine composition[J]. Analytica Chimica Acta, 2004, 513: 191–196.
[40] Soufleros E H, Bouloumpasi E, Tsarchopoulos C, Biliaderis C G. Primary amino acid profiles of Greek white wines and their use in classification according to variety, origin and vintage[J]. Food Chemistry, 2003, 80: 261-273.
[41] Juhász O, and T?rley D. Effect of Free Amino Acids of the Grape on the Development of Organoleptic Properties of Wine[J]. Acta Aliment,1985, 14(2): 101-112.
[42] Kiralp S, and Toppare L . Polyphenol content in selected Turkish wines, an alternative method of detection of phenolics[J]. Process Biochemistry,2005, 29: 125-129.
[43] Schwarz M, Hillebrand S, Habben S, Degenhardt A, Winterhalter P. Application of high-speed countercurrent chromatography to the large-scale isolation of anthocyanins[J]. Biochemical Engineering Journal, 2003,14:179-189.
[44] Kallithraka S,Arvanitoyannis I, El-Zajoulia A, and Kefalas P. The application of an improved method for trans-resveratrol to determine the origin of Greek red wines[J]. Food Chemistry,2001,75:355–363.
[45] ]Mu?oz-Espada A C, Wood K V, and Bordelon B. Anthocyanin Quantification and Radical Scavenging Capacity of Concord, Norton, and Marechal Foch Grapes and Wines[J]. Journal of Agricultural and Food Chemistry, 2004, 52: 6779-6786.
[46] Burns J, Mullen W, and Landrault N. Variations in the Profile and Content of Anthocyanins in Wines Made from Cabernet Sauvignon and Hybrid Grape[J]s. Journal of Agricultural and Food Chemistry, 2002,50:4096-4102.
[47] Gómez-M?guez M and Francisco J. H. Effect of the Maceration Technique on the Relationships between Anthocyanin Composition and Objective Color of Syrah Wines[J]. Journal of Agricultural and Food Chemistry, 2004, 52:5117-5123.
[48] Villiers A, Vanhoenacker G, Majek P, Sandra P . Determination of anthocyanins in wine by direct injection liquid chromatography–diode array detection–mass spectrometry and classification of wines using discriminant analysis[J]. Journal of Chromatography A, 2004, 1054:195-204.
[49] Brouillard R, Chassaing S, and Fougerousse A. Why are grape/fresh wine anthocyanins so simple and why is it that red wine color lasts so long[J]. Phytochemistry, 2003,64:1179-1186.
[50] Berente B , Garc?a D C, Reichenbacher M, and Klaus D. Method development for the determination of anthocyanins in red wines by high-performance liquid chromatography and classification of German red wines by means of multivariate statistical methods[J]. Journal of Chromatography A,2000, 871:95-103.
[51] Darias-Martín J, Carrillo M, and Díaz E. Enhancement of red wine colour by pre-fermentation addition of copigmentsp[J]. Food Chemistry, 2001,73:217-220.
[52] Nú?ez V, Monagas M, Gomez-Cordovés M C, and. BartoloméB. Vitis vinifera L. cv. Graciano grapes characterized by its anthocyanin profile[J]. Postharvest Biology and Technology,2004,31:69-79.
[53]丛浦珠,苏克蔓.化学手册(第2版)第九组质谱分析[M ].北京:化学工业出版社, 2000:27-51.
[54] Vázquez L C, Pérez-Coello M S, and Cabezudo M D. Effects of enzyme treatment and skin extraction on varietal volatiles in Spanish wines made from Chardonnay, Muscat, Airén, and Macabeo grapes[J]. Analytica Chimica Acta. 2002, 458: 39-44.
[55]李记明,贺普超.中国野生葡萄酒风味成分分析.果树学报,2004,21(1):11-16.
[56] Isabelle S. Gilles R, Yorgos K,and. Alain B. Determination of Volatile Compounds in Grenache Wines in Relation with Different Terroirs in the Rhone Valley[J]. Journal of Agricultural and Food Chemistry, 2002,50,6341-6345.
[57] Selli S, Canbas A, Cabaroglu T, Huseyin E., Ziya G. Aroma components of cv. Muscat of Bornova wines and influence of skin contact treatment[J].Food Chemistry,2006,94:319-326.
[58] Calleja A and FalquéE. Volatile composition of Mencía wines[J]. Food Chemistry, 2005,90:357-363.
[59] Peinado R A, Moreno J A, Mu?oz D, Manuel M. and Juan M. Gas Chromatographic Quantification of Major Volatile Compounds and Polyols in Wine by Direct Injection[J]. Journal of Agricultural and Food Chemistry, 2004,52:6389-6393.
[60] Sabon I, Derevel G, Kotseridis Y, and Bertrand A . Determination of Volatile Compounds in Grenache Wines in Relation with Different Terroirs in the Rhone Valle[J]. Journal of Agricultural and Food Chemistry, 2002, 50:6341-6345.
[61] Boido E, Lloret A, and Medina K. Aroma Composition of Vitis vinifera Cv. Tannat the Typical Red Wine from Uruguay[J]. Journal of Agricultural and Food Chemistry, 2003,51:5408-5413.
[62] FalquéE, Fernández E and Dubourdieu D. Volatile Components of Loureira, Dona Branca, and Treixadura wines[J]. Journal of Agricultural and Food Chemistry, 2002, 50:538-543.
[63] Pazourek J, Gajdo?ováD, SpaniláM, FarkováM, NovotnáK, Havel J. Analysis of polyphenols in wines: Correlation between total polyphenolic content and antioxidant potential from photometric measurements Prediction of cultivars and vintage from capillary zone electrophoresis fingerprints using artificial neural network[J]. Journal of Chromatography A,2005,951:98-104.
[64]李华.现代葡萄酒工艺学[M].陕西人民出版社,2000.
[65]国家药品监督管理局.中药注射剂指纹图谱研究的技术要求(暂行) [S].中成药,2000,20(10):671-675.
[66] Mateus N, Silva A M S, and Vercauteren J. Occurrence of anthocyanin-derived pigment in red wines[J]. Journal of Agricultural and Food Chemistry, 2001,49: 4836-4840.
[67] García-Beneytez E, Cabello F, and Revilla E. Analysis of Grape and Wine Anthocyanins by HPLC-MS[J]. Journal of Agricultural and Food Chemistry, 2003, 51:5622-5629.
[68] García-Beneytez E, Revilla E, and Cabello F. Anthocyanin pattern of several red grape cultivars and wines made from them[J]. European Food Research and Technology,2002,215:32-37.
[69]曾志,杨东辉,宋力飞等.高效液相色谱指纹图谱应用于板蓝根的鉴定[J].分析化学,2002,30(7):849-852.
[70]卢纹岱.SPSS for windows统计分析[M].北京,电子工业出版社,2004,311-327.
[71] Tsanova-Savova S, Dimovw S and Ribarova F. Anthocyanins and Color Variables of Bulgarian Aged Red Wines[J]. Journal of Food Composition and Analysis, 2002,15: 647-654.
[72]张军翔,冯长根,李华.不同品种红葡萄酒花色苷高效液相色谱指纹图谱识别[J],农业工程学报, 2006,22(10):216-221.
[73] Bridle P and Garcia-Viguera C. A simple technique for the detection of red wine adulteration with elderberry pigments[J].Food Chemistry,1996,55(2):111-l13.