Comparison of Sangiovese and Cabernet Franc Wines Made in Amphora and Stainless Steel

1. Comparison of the chemical/sensorialcharacteristics of Sangiovese and Cabernet Francwines made in amphora and stainlesssteel Tania Martellini Department of Chemistry «Ugo Schiff» – University of Florence Terracotta and Wine 2018 - Antica Fornace Agresti, Impruneta, 24-25 November2018

2. Preview of the project → Terracotta and Wine 2016 "Influence of fermentation and refining in amphora on the characteristics of wine»

3. In collaboration with: MULTIDISCIPLINARY RESEARCH TEAM Department of Chemistry «Ugo Schiff» – University of Florence Laboratory di Microanalisi di Agraria- DISPAA- Università degli Studi di Firenze Consorzio interuniversitario per lo Sviluppo dei sistemi a Grande Interfase (CSGI) – Firenze Centro di Ricerca di Risonanze Magnetiche (CERM) – Firenze Analytical FOOD - Firenze "Influence of fermentation and refining in amphora on the characteristics of wine»

4. Why use terracotta? • Image linked to oldtraditions • Use of a material of naturalorigin • Craftsmanship Characterization of the image of a company • Permeability of oxygensimilar to that of wood (micro-oxygenation) • Goodthermalinsulation • Minimum aromatictransfer to the winecompared to wood(respecting the wine’svarietalcharacteristics.) Interestingoenologicaltool

5. Aim of the Research Studyingthe potentialitiesand limits of vinification in amphora with a scientificapproach: • Evaluation of the influence of the firingtemperature of the amphora on the sensorialproperties of the wine • Evaluation of the influence of the micro-oxygenation of the amphora on the wine by monitoring the characteristics of porosity of the terracotta during the winemakingprocess • Evaluation of the effectivecontribution of terracotta to the aromatic and mineralprofile of the wineusinga stainlesssteelsteel container ("witness") for comparison.

6. The Project Clay from Impruneta «Light» Amphora T= 900°C ; t= 78h «Dark» Amphora T= 1050 °C ; t= 70h

7. Cabernet Sangiovese The Project «Dark» Amphora «Dark» Amphora «Light» Amphora «Light» Amphora Steel Steel 2016 Vintage Post-Fermentation (end of October2016) Mid-Process (earlyMay 2017) End-Process (end of June 2017)

8. Instruments and methods: analysisof winesamples SpectometerNMR BrukerBioSpin 400 MHz (CERM) ICP-OES Optima 2000 PerkinElmer (DISPAA) Analisis of metals via ICP-OES Analisisof the aromaticcomponents of the vino via HS-SPME-GC-MS NMR Analisis New metabolomicapproachwith winesamples Gas chromatographTrace GC + Trace DSQ ThermoFinnigan + CTC Analytics CombiPAL (AnalyticalFood)

9. Materials and methods: study of the characteristics of the amphora SAMPLING: 1.Before vinification 2.After first racking (October 2016) 3.After end of process(June 2017) Gas-porosimetry Capillaryascent Water vaporpermeability Dryingkinetics 𝑪𝑨=𝒂/√𝒕∗ Non-destructiveanalysis: preliminary information Gas-porosimeterCoulter SA 3100 (CSGI)

10. Materials and methods: study of the characteristics of the amphora SAMPLING: 1.Before vinification 2.After first racking (October 2016) 3.After end of process(June 2017) Samples from Jar post-vinification Averagecapillaryabsorptioncoefficient 0,005-0,006 g/cm2s1/2 Averagecapillaryabsorptioncoefficient 0,001 g/cm2s1/2 Dark Cabernet Dark Sangiovese Reduction of the coefficient → Manifestation of a change in the chemical-physicalcharacteristicsrelated to the porosity of the material Light Cabernet Light Sangiovese

11. Materials and methods: study of the characteristics of the amphora SAMPLING: 1.Before vinification 2.After first racking (October 2016) 3.After end of process(June 2017) Samples from Jar post-vinification The dryingkinetics and permeability to water vaporconfirmswhatisdetermined with the capillaryascent Dark Cabernet Dark Sangiovese Markeddifferencebetween the permeability of the jaraftervinification with Sangiovese and that with Cabernet Light Cabernet Light Sangiovese

12. Gas-porosimetry: specific area measurement (BET) MONOLAYER FORMATION MULTILAYER FORMATION Volume absorbed CAPILLARY CONDENSATION MICROPORE FILLING • BET methoddeveloped by Brauner, Emmett and Teller • Solid surface area correlated to a gas thatisadsorbedsuperficially • Typicalisotherms of the mesopores (150-2nm) with hysteresis • The hysteresis and the BET increasein particular for the Cabernet vinified in the light amphora

13. Gas-porosimetry: measurement of specific area (BET) Metodo BET multilayerabsorbtion: Information on the porosity of the material (specific area m2/g) from the volume of nitrogenabsorbed from the surface of the sample. DestructiveTecniques Coulter Gas-porosimeter SA 3100 (CSGI) Higher BET values => the residue of winegetinto the porescreating a surface Similar BET values Pre-vinification First Racking Post-Vinification BET valuessuperior in the light amphora => more porous More pronouncedeffect with Cabernet

14. Gas-porosimetria: misura di area specifica (BET) BET Method multilayerabsorbtion : Information on the porosity of the material (specific area m2/g) from the volume of nitrogenabsorbed from the surface of the sample. DestructiveTecniques Gas-porosimetroCoulter SA 3100 (CSGI) • BET valuesPre-vinification → higher for the more porouslight amphora • BET valuesBeforeracking and pre-vinification → are similar, rackingdoesnotaffectthem • BET valuesHigher post-aging → the wine residue getsintothe pores, creating a surfacedevelopment • Pronouncedeffectaboveall for Cabernet vinified in the light amphora

15. Aroma Analysis Extraction in solidform from the top HS-SPME-GC-MS Fibra SPME Supelco 50/30 µm DVB/Carboxen/PDMS SatbleFlex Method of acquisition: Full Scanof mass rangefrom 30 to 350 Relative Area OverThreshold [Zhuet al 2015] Area of compositionin % of totaldivided by relative Odor Detection Threshold

16. Aroma Analysis: Post Fermentation(October 2016) Notmuchdifferencebetweensteel and amphora Common varietalaromas: Terpenes and Norisoprenoids (floreal) LowODT Aroma Enhancer β- Damascenone Linalol β- Citronellol SecondaryAromas (fermentative): estersalcohols and acids (fruity notes) Ethyloctanoate Ethylbutanoate Isoamyl acetate Ethylhexanoate (caporato) OnlyCabernet OnlySangiovese α- terpineol Methyloctanoate

17. Aroma analysis: Mid-Process(May 2017) • Lesserdifined RAOT trends productstill in phase of evolution • Sangiovese has a greaternumber of compounds (butyricacid and fufural SOUR, BITTER and TOASTED • Fatty Acid Aroma Description • octanoic acid almond butter • non -anoic acid herbaceous, fat • decanoic acid rancid • dodecanoic acid metallic Fatty AcidsAroma Descriptions Hexanoicacid Grassy, Woody Octanoicacid Almond butter Non-anoicacid Herbaceous, fatty Decanoicacid Rancid Dodecanoicacid Metallic Presence of short-chainalcohols (pungentodor) and fattyacidssuccessivelyalienatedduringwinemakingprocess

18. Aroma Analysis: Post end of process(June 2017) • Increases the number of aromas (esters) • Higher RAOT for varietalaromas and esters Hexanoicacid (GRASSY) Phenylethanol(PEONY) “CULTIVAR DIPENDENZA” • RAOT are higher for: • Volatile phenols: 4-ethyl guaiacol and 4 ethyl-phenol No substantialdifferencebetween Light and dark amphorae Welldefined RAOT trend Greater differencebetween the RAOT in amphora and steel in the Cabernet comparedwith Sangiovese The Aromaticprofile of Cabernet is more enhanced in amphora

19. Analysis of metals The totalcontent of metalsbecomesgreaterduring the winemakingprocess Little difference in the wine made in light and dark amphorae Steel Light Amphora Dark Amphora Steel Light Amphora Dark Amphora Post fermentationMid– Termprocess End process Per il Cabernet: Inferiortotalcontent of metals in anfora compared to steel The contribution of micro-oxygenation in cabernet in amphorawas more evidentcomparedto Sangiovese

20. Analysis of metals : micro-oxygenation Steel Light Amphora Dark Amphora Trend of Sulphur: Example of the micro-oxygenation in Amphora Mid-Term process End process Steel Light Amphora Dark Amphora Mid-Term process End process

21. NMR Analyses Wine Profiling Report Metabolitesquantified with the Bruker Wine Profilingtool • formicacid • Methanol • 2,3 butanediol • 2-phenylethanol • 3-methyl butanol • Acetalde • hydegalacturonic acid • succinic acid • glycerol/ ethanol • Proline • caftaric acid • Epicatechin • Gallic acid • shikimic acid • trigonellina • totalalcohol • Table • Ethanol • Ethanolvol. • Glycerol • Fructose • Arabinose • totalsugars (beforeinversion) • non-reducingsugars • tartaric acid • malic acid • lactic acid • energyvalue • aceticacetoin acid • ethylacetate The analyses of the metabolitesconfirm the valuesobtained from routine analysis on the wine made with classicalmethods

22. NMR analysis: statistical treatment PCA (Principal Component Analysis) • Discriminationbetweenamphora and steel: verygood (accuracyof 87.5%) • Discriminationbetween light amphora and steel and between dark amphora and steel: verygood(accuracy 86.6% and 92.3%) • Poordiscriminationbetween light amphora and dark amphora (accuracy 15.4%) OPLS (OrthogonalProjections to LatentStructures) Amphora vs Steel Light Amphora vs Dark Amphora

23. Conclusions: Effects on porosity • First rackingdoesnotaffect the porosity of the amphora • Variationof porosityin the amphoraappearsafterrefinement, especially with Cabernet • Micro-oxygenationseemsto have a greatereffect on Cabernet compared to Sangiovese

24. Conclusions: Effects on the AromaticCharacteristicsof the Wine • The firing temperature of the amphoradoesnotsignificantlyinfluence the aromatic, mineral and metabolomicprofile of the wine • The composition of the wineafter the first racking (October 2016) and Mid-termprocess (May2017) shows no differencesbetweenamphora and steel • Importantdifferences are notedbetweenamphora and steelafter the end of process(June 2017) which are dependenton cultivars

25. THANK YOU TO EVERYONE Wethankallthosewhoparticipated in the realization of thisresearch In particular Cincinelli, L. Sposato, C. Guerranti, C. Marinelli, M. Fortini, R. Giorgi, S. Pucci, A. Ciaccio, L. Tenori, G. Meoni, F. Bartoletti, C. Luchinat, P. Baglioni, L. Lepri Il Castello del Trebbio L. Parisi - Artenova

26. Grazie per l’attenzione!!!!!!

28. NMR Analyses

29. Analisi NMR The NMR Report isquitecomplex…..only a fewparts…..

30. Il progetto