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The Impact of Commercial Tannin Additions and Addition Timing on Finger Lakes Red Wine Color A Thesis Presented to the Faculty of the Graduate School of Cornell University in Partial Fulfillment of the Requirements for the Degree of Master of Science by Robert George Gallasch August 2012  2012 Robert George Gallasch ii Abstract In winemaking, the addition of enological tannins is intended to improve color extraction, color stability during aging, or various sensory properties. Many different commercial tannin preparations are available to winemakers, but caution in selection and use are called for, as product composition can vary by batch, manufacturer, plant source and extraction method used. Likewise, little definitive guidance on use or addition rate is available to winemakers beyond manufacturer recommendations or anecdotes from other winemakers. This experiment was designed to determine whether the same relative dose of tannin material added at different times—before, during or after fermentation—affected wine color development and stability after two years of bottle aging. Red wines were made from Finger Lakes Pinot Noir and Lemberger, two cultivars with relatively low tannin content, and treated with two different types of commercial tannins. Samples were analyzed by direct spectrophotometric readings, protein precipitation assays and high performance liquid chromatography (HPLC) to assess differences in phenolic fractions, co-pigmentation factors and visible color characteristics. Analysis revealed few significant differences between treatments. Variations occurred during fermentation, and some significant differences were noted after three months in the bottle; but few significant differences were apparent after two years. Keywords: tannin, enological tannin, additives, anthocyanin, copigmentation, red wine color, Pinot Noir, Lemberger, wine aging, polymeric pigments, LPP, SPP iii Biographical Sketch Major Robert G. Gallasch, USAF Retired, earned his Bachelor of Arts degree in History in 1984 from the University of Rochester, Rochester NY, and a Masters of Arts degree in International Relations from Creighton University, Omaha NE, in 1999. After twenty years of service, he retired from the United States Air Force and then attended Cornell University to study viticulture and wine making. He lives with his family on a small farm in Moravia, New York. iv Dedication This work is dedicated to my family. My beloved wife and partner in life, Gail; and my wonderful children, Robert and Samantha. Without their patience, love and support this project could never have been undertaken, much less completed. v Acknowledgements Cornell University, Department of Food Science Enology Extension Lab, NYSAES, Geneva NY Vinification & Brewing Lab, NYSAES, Geneva NY Dr. Anna Katharine Mansfield Dr. Gavin L. Sacks Dr. David C. Manns Mr. Chris Gerling, Ms. Rebecca Nelson, Mr. Mark Nisbet, Ms. Céline Coquard- Lenerz, Mr. Ben Gavitt, Ms. Luann Preston-Wilsey, Ms. Pamela Raes Cornell Orchards, Lansing NY Sheldrake Point Winery, Ovid NY The New York Farm Viability Institute United States Department of Veterans Affairs vi Table of Contents Abstract ......................................................................................................................................................................................... iii Biographical Sketch ................................................................................................................................................................. iv Dedication ..................................................................................................................................................................................... v Acknowledgements .................................................................................................................................................................. vi Table of Contents .................................................................................................................................................................... vii List of Figures .............................................................................................................................................................................. x List of Tables ............................................................................................................................................................................... xi Literature Review ................................................................................................................... 1 Introduction ................................................................................................................................................................................. 1 Wine Color Chemistry .............................................................................................................................................................. 3 Phenolics ........................................................................................................................................................................................ 3 Anthocyanins ............................................................................................................................................................................... 5 Co-‐pigmentation ......................................................................................................................................................................... 6 Tannins ........................................................................................................................................................................................ 12 Role of Tannins in Wine ....................................................................................................................................................... 13 Hydrolysable Tannins ........................................................................................................................................................... 13 Condensed Tannins ................................................................................................................................................................ 14 Analysis of Phenolic Compounds ..................................................................................................................................... 16 Associative assessments of wine ...................................................................................................................................... 16 Early instrumental analysis of wine tannin and color ............................................................................................ 17 Direct Spectrophotometric Methods .............................................................................................................................. 18 Photometric methods (direct measurement of color) ............................................................................................. 18 Oxidation-‐Precipitation methods .................................................................................................................................... 18 Indirect measurement of phenolic fractions ............................................................................................................... 18 Folin-‐Denis method (total phenolics) ............................................................................................................................. 19 Folin-‐Ciocalteu method (total phenolic content) ...................................................................................................... 19 Current spectrophotometric methods ........................................................................................................................... 20 High Performance Liquid Chromatography (HPLC) ................................................................................................ 23 Polymeric nature of wine pigments ................................................................................................................................ 24 Measuring polymeric pigments ......................................................................................................................................... 24 Measuring anthocyanins (monomeric pigments and copigmentation) .......................................................... 25 Pyranoanthocyanin color compounds ............................................................................................................................ 27 Protein Precipitation methods .......................................................................................................................................... 29 Assessing total tannins and tannin fractions .............................................................................................................. 29 Iron-‐reactive phenolics method ........................................................................................................................................ 31 Polysaccharide (methyl cellulose) precipitation methods ................................................................................... 31 Improving phenolic extraction from grapes ................................................................................................................ 32 Phenolic concentration by cultivar ........................................................................................................................................... 32 Maceration effects ............................................................................................................................................................................ 33 Aging ...................................................................................................................................................................................................... 34 Cap management .............................................................................................................................................................................. 34 Combination techniques ............................................................................................................................................................... 35 Enological tannin additions ................................................................................................................................................ 35 Use and availability of enological tannins .................................................................................................................... 35 Types of commercial enological tannin products ..................................................................................................... 37 The effect of tannin additions on color formation and stability ......................................................................... 37 Impact of fruit quality on the tannin addition efficacy ........................................................................................... 40 vii Composition of enological tannins ................................................................................................................................... 40 Sensory impact of tannin additions ................................................................................................................................. 43 Negative impact on wine color ................................................................................................................................................... 43 Impact on taste (sweetness, sourness, bitterness, and astringency) ........................................................................ 44 Impact on wine aroma ................................................................................................................................................................... 45 Chapter 2 .............................................................................................................................. 47 Introduction .............................................................................................................................................................................. 47 Materials and Methods ......................................................................................................................................................... 49 Grapes ........................................................................................................................................................................................... 50 Direct Spectroscopic Measurement ................................................................................................................................. 53 Phenolic analysis ...................................................................................................................................................................... 54 Reagents ...................................................................................................................................................................................... 56 Sample preparation ................................................................................................................................................................ 57 Calculations ................................................................................................................................................................................ 57 Statistics ...................................................................................................................................................................................... 57 Results and Discussion ......................................................................................................................................................... 60 Wine chemistry ......................................................................................................................................................................... 60 Varietal differences during fermentation ..................................................................................................................... 60 HPLC results ....................................................................................................................................................................................... 61 Phenolic and color development, during fermentation .......................................................................................... 61 Pinot Noir, during fermentation. ............................................................................................................................................... 61 Lemberger, during fermentation ............................................................................................................................................... 63 Phenolic and color development, post-‐fermentation ............................................................................................... 64 Pinot Noir, post-‐fermentation. .................................................................................................................................................... 64 Volutan initial treatment, Pinot Noir, post-‐fermentation. ........................................................................................ 65 Volutan incremental treatment, Pinot Noir, post-‐fermentation. ........................................................................... 66 Volutan final treatment, Pinot Noir, post-‐fermentation. ........................................................................................... 66 SR-‐Terroir initial treatment, Pinot Noir, post-‐fermentation. .................................................................................. 67 SR-‐Terroir incremental treatment, Pinot Noir, post-‐fermentation. ..................................................................... 68 SR-‐Terroir final treatment, Pinot Noir, post-‐fermentation. ..................................................................................... 69 Pinot Noir, Summary ................................................................................................................................................................. 69 Lemberger, post-‐fermentation ................................................................................................................................................... 70 Volutan initial treatment, Lemberger, post-‐fermentation. ....................................................................................... 71 Volutan incremental treatment, Lemberger, post-‐fermentation. .......................................................................... 72 Volutan final treatment, Lemberger, post-‐fermentation. .......................................................................................... 73 SR-‐Terroir initial treatment, Lemberger, post-‐fermentation. ................................................................................. 73 SR-‐Terroir incremental treatment, Lemberger, post-‐fermentation. .................................................................... 73 SR-‐Terroir final treatment, Lemberger, post-‐fermentation. .................................................................................... 74 Lemberger, Summary ............................................................................................................................................................... 75 Sensory screening results. ............................................................................................................................................................ 75 Practical Applications ........................................................................................................................................................... 77 Chapter 3: Future Work ........................................................................................................ 81 After-‐Action: Experiment Design Changes .................................................................................................................. 81 Streamlined Experiment Design ....................................................................................................................................... 81 Methodology Issues ............................................................................................................................................................... 82 Recommendations ................................................................................................................................................................... 84 Modifications to direct spectrophotometry ................................................................................................................. 85 Readings at 280 nm ................................................................................................................................................................ 86 Somers-‐Boulton Method ....................................................................................................................................................... 87 Adams-‐Harbertson .................................................................................................................................................................. 88 HPLC .............................................................................................................................................................................................. 88 viii Dose Rate Selection ................................................................................................................................................................. 89 Sampling Protocol ................................................................................................................................................................... 89 Equipment Issues ..................................................................................................................................................................... 90 Sensory Trials ............................................................................................................................................................................ 91 Future Research Projects .................................................................................................................................................... 91 Methodology improvement ................................................................................................................................................. 91 Commercial tannin characterization by HPLC ........................................................................................................... 91 Copigmentation experiment ............................................................................................................................................... 92 High Dose Rate Trials ............................................................................................................................................................ 92 Initial vs. Final Addition Trial ............................................................................................................................................ 93 Sensory Evaluation during aging ..................................................................................................................................... 93 Aroma Trials .............................................................................................................................................................................. 93 Tables ................................................................................................................................... 94 Figures ................................................................................................................................ 104 General ...................................................................................................................................................................................... 104 Pinot Noir ................................................................................................................................................................................. 107 Lemberger ................................................................................................................................................................................ 121 ix List of Figures Chapter 1: Figure 1-‐1: Simple Phenol ................................................................................................................................................................................. 3 Figure 1-‐2: Hydroxycinnamic Acid ............................................................................................................................................................... 4 Figure 1-‐3: Anthocyanidins ............................................................................................................................................................................... 4 Figure 1-‐4: Flavonoid Example (Malvidin) ................................................................................................................................................ 4 Figure 1-‐5: Flavonoid Ring System ................................................................................................................................................................ 5 Figure 1-‐6: Anthocyanins ................................................................................................................................................................................. 5 Figure 1-‐7: Hydrolysable Tannin sub-‐units ............................................................................................................................................ 12 Figure 1-‐8: An Ellagitannin ........................................................................................................................................................................... 12 Figure 1-‐09: Flavan-‐3-‐ol Structure ............................................................................................................................................................ 13 Figure 1-‐10: Pyranoanthocyanin ………………………………………………………………………………………………………………… ... 24 Chapter 2: Figure 2-‐1: Vineyard locations on Cayuga Lake, NY ......................................................................................................................... 110 Figure 2-‐2: Varietal differences during fermentation ..................................................................................................................... 111 Figure 2-‐3: HPLC trace comparison ......................................................................................................................................................... 112 Figure 2-‐P–1: Pinot Noir, visible color .................................................................................................................................................... 107 Figure 2-‐P–2: Pinot Noir, blueness ........................................................................................................................................................... 108 Figure 2-‐P–3: Pinot Noir, hue ..................................................................................................................................................................... 109 Figure 2-‐P–4: Pinot Noir, Visible Anthocyanins .................................................................................................................................. 110 Figure 2-‐P-‐5: Pinot Noir, color density ................................................................................................................................................... 111 Figure 2-‐P–6: Pinot Noir, color due to copigmentation .................................................................................................................. 112 Figure 2-‐P–7: Pinot Noir, polymeric pigments .................................................................................................................................... 113 Figure 2-‐P-‐8: Pinot Noir, total phenolics by direct spectrophotometry ................................................................................... 114 Figure 2-‐P–9:Pinot Noir, Total Phenolics by Folin-‐Ciocalteau ..................................................................................................... 115 Figure 2-‐P–10: Pinot Noir, Iron-‐reactive tannins .............................................................................................................................. 116 Figure 2-‐P–11: Pinot Noir, large polymeric pigments ..................................................................................................................... 117 Figure 2-‐P–12: Pinot Noir, small polymeric pigments ..................................................................................................................... 118 Figure 2-‐P–13: Pinot Noir, monomeric pigments .............................................................................................................................. 119 Figure 2-‐P–14: Pinot Noir, total polymeric pigments (color at pH 4.9) ................................................................................... 120 Figure 2-‐L-‐1: Lemberger, visible color .................................................................................................................................................... 121 Figure 2–L–2: Lemberger, blueness ......................................................................................................................................................... 122 Figure 2–L–3: Lemberger, Hue ................................................................................................................................................................... 123 Figure 2–L–4: Lemberger, visible anthocyanins ................................................................................................................................. 124 Figure 2–L–5: Lemberger, color density ................................................................................................................................................ 125 Figure 2–L–6: Lemberger, color due to copigmentation ................................................................................................................ 126 Figure 2–L–7: Lemberger, polymeric pigments .................................................................................................................................. 127 Figure 2–L–8: Lemberger, total phenolics by direct spectrophotometry ................................................................................ 128 Figure 2–L–9: Lemberger, total phenolics by Folin-‐Ciocalteau ................................................................................................... 129 Figure 2–L–10: Lemberger, Iron Reactive Tannins ........................................................................................................................... 130 Figure 2–L–11: Lemberger, large polymeric pigments ................................................................................................................... 131 Figure 2–L–12: Lemberger, small polymeric pigments ................................................................................................................... 132 Figure 2–L–13: Lemberger, monomeric pigments ............................................................................................................................ 133 Figure 2–L–14: Lemberger, total color at pH 4.9 .............................................................................................................................. 134 x

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by Robert Gallasch

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