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INSTITUTO NACIONAL DE VITIVINICULTURA

INSTITUTO NACIONAL DE VITIVINICULTURA. www.inv.gov.ar. RECOGNITION OF THE GEOGRAPHICAL ORIGIN OF ARGENTINE WINES VARIETAL CLASSIFICATION LINES OF WORK. LINEAS DE TRABAJO. INVESTIGACION. CONTROL. CARACTERIZACION VARIETAL. ORIGEN GEOGRAFICO. CERTIFICACION VARIETAL. ACREDITACION.

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INSTITUTO NACIONAL DE VITIVINICULTURA

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  1. INSTITUTO NACIONAL DE VITIVINICULTURA www.inv.gov.ar

  2. RECOGNITION OF THE GEOGRAPHICAL ORIGIN OF ARGENTINE WINES VARIETAL CLASSIFICATION LINES OF WORK

  3. LINEAS DE TRABAJO INVESTIGACION CONTROL CARACTERIZACION VARIETAL ORIGEN GEOGRAFICO CERTIFICACION VARIETAL ACREDITACION ORIGEN ALCOHOL Y AGUA ESPECTROMETRIA POLIFENOLES HPLC RELACION ISOTOPICA ESPECTROMETRIA ADN AROMAS GC CATIONES POR ICP AMINOACIDOS ANIONES INORGANICOS ANALISIS SENSORIAL ANALISIS MULTIVARIADO ANALISIS MULTIVARIADO RECOGNITION OF THE GEOGRAPHICAL ORIGIN OF ARGENTINE WINES - VARIETAL CLASSIFICATION PESTICIDAS DEFECTOS DEL VINO OTROS

  4. EQUIPMENT • HPLC with diode arraydetector • HPLC with UV Detector with double furnace • Inductively coupled Plasma- axial and radial observation atomic emission spectrometer • Gas Chromatograph with FID detector • Ionic Chromatograph with electrochemical detectors, of Amperometric and Conductimetric cells • Mass Spectrometers

  5. MALBEC TEMPRANILLO BONARDA SYRAH / SHIRAZ CABERNET SAUVIGNON TANNAT MERLOT CHARDONNAY SAUVIGNON BLANC RIESLING TORRONTES RIOJANO SEMILLON PINOT GRIS VARIETIES

  6. MICROVINIFICATIONS • Approximately 600 samples per year since 2004.

  7. HUMAN RESOURCES • Taking part of these lines of work: • 8 professionals • ALL OF THEM WITH POSTGRADUATE DEGREES • 3 Ph.D. Candidates

  8. TRAININGS • Geographic Origin - Laboratoire Tálense de la Direction Genérale de la Concurrence de la Consommation et de la Represión des Fraudes de Bordeaux-France, belonging to the Ministère de l'Économie, des Finances et de l'Industrie from the French Government • Varietal Differentiation - Institut fur Lebensmittelchemie, Trier, Germany and in Laboratory Experimentale D’Asti, Italy • “Latest Technologies in the Application of Stable Isotopes” -Eurofins Research Centre, Nantes, France and in Station Experimental Zaidin (CSIC), Granada, Spain • Genetic Origin Determination of Vine Varieties - INSTITUTO DE FERMENTACIONES INDUSTRIALES, belonging to the Consejo Superior de Investigaciones Científicas from the Ministerio de Educación y Ciencia in Spain (CICS).

  9. NEW CONTROL ANALYTICAL TECHNIQUES FOR WINE GEOGRAPHIC ORIGIN

  10. THE STUDIES ARE ORIENTED TOWARDS • WINE DESCRIPTION • BY MEANS OF ROUTINE ANALYSIS

  11. METHODOLOGY • COMPARISONS • ANALYSIS ON SAMPLES EXTRACTED IN WINERY • ANALYSIS ON SAMPLES OF FREE CIRCULATION REQUEST OR FRACTIONED SAMPLES

  12. ROUTINE ANALYSIS • METHANOL • ACETALDEHYDE • ETHYL ACETATE • PROPANOL • ISOBUTANOL • CHLORIDES • 13C/12C • PHOSPHATES

  13. ORIGINAL WINE

  14. EXPECTED EVOLUTION OF SAME WINE

  15. UNEXPECTED EVOLUTION

  16. THE STUDIES ARE ORIENTED TO: • DETERMINING THE GEOGRAPHICAL ORIGIN OF A WINE BY MEANS OF THE MULTIVARIED ANALYSIS OF CATIONS AND ANIONS

  17. DATA BANKExtraction of grape samples MENDOZA SAN JUAN LA RIOJA CATAMARCA SALTA RIO NEGRO CENTRE AND EAST OF THE COUNTRY

  18. CATIONS WHICH CONTRIBUTE TO “DETERMINE” THE WINE ORIGIN REGION Rb, Li, Cd, Pb, Mn, Mg, Zn, Ca, Na

  19. DISCRIMINATING ANALYSIS OF MENDOCINIAN DEPARTMENTS

  20. TUPUNGATO, LA PAZ AND SAN MARTIN DISCRIMINATING ANALYSES

  21. BY MEANS OF THE ROUTINE ANALYSIS GEOGRAPHIC ORIGIN CAN BE “CONTROLLED” BY MEANS OF THE DISCRIMINATING ANALYSIS OF CATIONS GEOGRAPHIC ORIGIN CAN BE “DETERMINED”

  22. VARIETAL CHARACTERIZATION VARIETAL WINE • Concept: it expresses the predominance of a variety in a wine as its fundamental component • Administratively it is defined asa minimal proportion, 85%, of a grape variety

  23. Red Wine Varietal Characterization

  24. Varietal Characterization Applying Chemical Methods • Anthocyanic monomer profile, shikimic acid profile, hydroxycinnamic acids linked to tartaric acid, resveratrol and flavonols, catequin and procyanidin profiles

  25. Red Wine Anthocyan Chromatogram

  26. Varietal Characterization Discriminating Analysis among Six Red Varieties

  27. Shikimic Acid Determination Wines by HPLC Sucrose (C6) ↕ Shikimic Acid ( C6-C1) ↨ Flavanes (C6-C3-C6) ↨ ↨ Tannins Anthocyan (C15)

  28. DETERMINATION OF PHENOLICCOMPOUNDS • 1-Galic Acid 6- Catequin 11- Epicatequin • 2-Protocatequic Acid 7- Vanillic Acid 12- Coumaric Acid • 3-Protocatequic Aldehyde 8- Caffeic Acid 13- Ferulic Acid • 4- Gentisic Acid 9- Syringic Acid 14- Sinamic Acid • 5-4-hydroxybenzaldehyde 10- Vanillin

  29. CONCLUSIONS • The application of chemical methods to determine the profile of anthocyans, hydroxycinnamic acid, resveratrol, shikimic acid, flavonols and flavanes is a powerful tool to ensure the varietal authenticity of wines. • Samples of unquestionable origin are required in order to obtain information and continue studying which are the variables that better fit the model to ensure characterization. • It is very important to take into account that data banks do not always reflect winery vinification and therefore it is necessary to consider those elements which vary of a microvinification with a large scale vinification. For that reason, the samples extracted in the winery will also be useful to correlate the results obtained with the INV data bank acquired from microvinifications.

  30. VARIETAL CHARACTERIZATION AMINO ACIDIC AND AROMATIC PROFILE Determination of Amino Acids by Anion-Exchange Chromatography with Integrated Pulsed Amperometric Detection (AAA-Direct™) High-performance anion-exchange chromatography resolves all common amino acids using NaOH and sodium acetate as eluyents. Integrated pulsed amperometric detection directly notices amino acids and carbohydrates.

  31. AMINO ACIDIC PROFILE • Studies indicate that free amino acids can be useful for the purposes of wine characterization (Vas Concelos and Das Neves, Chaves 1985). • Grape juice amino acids profile seems to be highly dependent on factors such as grape variety, soil type, and weather conditions (Flanzy and Poux, 1965).

  32. 80 70 60 50 40 30 20 10 0 0 5 10 15 20 25 30 35 40 45 Chromatogram of Amino Acid Standards Sample: 12–29 µM NIST amino acid standards with 25 µM tryptophan and norleucine Peaks: 1. Arginine (Arg) 6. Valine (Val) 11. Methionine (Met) 16. Glutamate (Glu) 2. Lysine (Lys) 7. Serine (Ser) 12. Norleucine (Norleu) 17. Aspartate (Asp) 3. Alanine (Ala) 8. Proline (Pro) 13. System 18. Cystine 4. Threonine (Thr) 9. Isoleucine (Ile) 14. Histidine (His) 19. Tyrosine (Tyr) 5. Glycine (Gly) 10. Leucine (Leu) 15. Phenylalanine (Phe) 20. Tryptophan (Trp) Eluent % 500 14 Deionized water 250 mM NaOH 1.0 M NaOAc 19 1 nC 18 16 15 2 20 17 4 13 5 7 10 3 11 9 8 6 12 0 Minutes

  33. Peaks: 1. Void 2.Arg 3. Lys 4. Gln 5. Asn 6. Ala 7. Thr 8. Gly 9. Val 10. Ser 11. Pro 12. Ile13. Leu14. Met15. System16. His17. Phe18. Glu19. Asp20. Cystine21. Tyr22. Trp 0 5 10 15 20 25 30 35 40 44 Minutes Perfil de Aminoacidos en Vino Moscatel 240 2 11 16 nC 1 17 6 5 10 7 19 3 8 13 4 18 9 15 20 14 12 22 21 30 22743

  34. AROMATIC PROFILE The aromatic profile analysis of 56 wineodorous components is carried out by means of solid-liquid extraction and subsequent concentration of volatile compounds, purification of extracts, identification and quantification.

  35. The 56 studied volatile compounds were classified according to sensory perception into: alcoholic, floral, fruity, herbaceous, lactic, vegetal, animal, resinous and spicy. The obtained results were statistically processed with the Package Statgraphics plus 6.0 so as to find ponderable relations between the volatile composition and concentration with the variety and geographic origin.

  36. 2-methyl-1-propanol 3-buten-1-ol 1-butanol ethyl propanoate acetal 2-methyl-1-butanol 3-methyl-1-butanol 4-methyl-2-pentanol 1-pentanol 3-methyl-1-butanol acetate ethyl butanoate Buthyl acetate + ethyl lactate propanal E-3-hexenol Z-3-hexenol E-2-hexenol hexanol 3-methyl-1-butanol acetate Pentyl acetate Gamma-butyrolactone Alpha-pinene ethylhexanoate Hexyl acetate D-limonene Cineol Benzylic alcohol Gamma-terpinene Guaiacol 3-methoxy-2-isopropilpirazin Linalol Alpha-terpinene 2-phenylethanol AROMATIC PROFILE

  37. 1-phenyl-2-propanol + E-limonene oxide Z-limonene oxide Citronelal 4-terpineol 4-ethylphenol 2-isobutyl-3-metoxipirazin Diethyl fumarate Ethyl octanoate Nerol Citronelol Neral Myrtanol Geraniol + linalyl acetate Z-cinnamaldehyde Geranial Decanol Methyl anthranilate Eugenol Neryl acetate Geranyl acetate Ethyl decanoate Ethyl anthranilate Alpha-ionone Beta-ionone S-nerolidol R-nerolidol Ethyl laurate AROMATIC PROFILE

  38. STUDY PROGRESS • 18 samples of 5 Vitis vinifera varieties have been analysed in duplicate: Syrah, Cabernet Sauvignon, Tempranillo, Merlot and Bonarda, from different geographical regions and different vintages.

  39. DISCRIMINATING ANALYSIS • The objective was to show the varietal differences among the five groups (Merlot, Cabernet, Syrah, Tempranillo and Bonarda) and the geographical differences among the groups (Tupungato, Junin, Tunuyán and San Juan), by means of the “discriminating analysis”, using the determinations of the 56 aromatic compounds in each sample.

  40. CONCLUSIONS • By means of this study, significant differences have been established in the tested samples among varieties and geographical regions with regard to the group of measured variables (volatile compounds), and the systematic classification for new determinations of samples of unknown origin is explained.

  41. Plot of Discriminant Functions 4.3 GEOGRAPHIC ORIGIN JUNIN LUJAN 2.3 SAN JUAN TUNUYAN 0.3 TUPUNGATO Centroids Function 2 -1.7 -3.7 -6 -3 0 3 6 9 12 Function 1

  42. ¡ M U C H A S G R A C I A S ! INSTITUTO NACIONAL DE VITIVINICULTURA Sede Central: Av. San Martín 430 (5500) – Mendoza – REPUBLICA ARGENTINA Tel/Fax (0261) 521-6604/06 www.inv.gov.ar

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