Understanding and manipulating Pinot noir phenolic profiles Bob Dambergs
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1 Understanding and manipulating Pinot noir phenolic profiles Bob Dambergs Marlborough 17 June 2014
2 The history of Pinot Noir Pinot noir is an ancient variety that may be only one or two generations removed from wild vines The name is thought to relate its cone shaped bunches- pineon - pine cone First positive description was from when the Romans arrived in Gaul - the vines were already there Some thought that the original domestication of wild vines that became Pinot Noir occurred in what is now Burgundy
3 A preference for Pinot Noir is in our primate genes Baboons at the La Terra de Luc vineyard near Cape Town prefer Pinot to other grape varieties!
4 Pinot noir mutates if you give it a cross look.marq De Villiers Hundreds of clones - related to its age and selection pressure over the years? Pinot Meunier Pinot Gris Pinot Blanc
5 Some of the great wines of the world are made with Pinot Noir, but..
6 The downside of Pinot Noir Climate specific Shoots early then grows slowly, so it s a frost risk Poor fruit set Thin skinned Susceptible to downy mildew, powdery mildew, botrytis, viruses Needs exposure for colour, but burns easily Low anthocyanin concentrations and anthocyanins are unstable Grapes have high tannin but it is mostly seed tannin Wines have relatively low tannin and colour.etc, etc
7 What defines a red wine? Chateau La Bob 1952 Water Ethanol Organic acids, glycerol Phenolics - tannins, anthocyanins Minerals Flavour compounds
8 Phenolic maturation profiles in grapes Stem Seed Skin Robinson and Walker, 2006
9 Anthocyanin facts Anthocyanins normally have a glucose molecule attached via a glycosidic bond, making them more stable.. GG Anthocyanins also exist as acetyl and courmaryl derivatives which are even more stable At very low ph (<2) all anthocyanins have a positive charge and are coloured, at wine ph only a small proportion are charged If SO 2 binds to anthocyanins they are colourless React with tannin to form stable pigments
10 Some tannin facts Pigmented tannin colour is less sensitive to ph Pigmented tannin colour is not bleached by SO 2 Non-bleachable pigments Tannins bind to proteins, to produce astringency, drying chalky characters when they bind to saliva linings of the mouth.their sensory effect is tactile rather than taste Seeds tannins vary in structure to skin tannin so their chemical reactivity and sensory properties vary Seed tannin is more difficult to extract than skin tannin
11 High seed tannin does not mean high wine tannin High skin tannin grapes Wine tannin (mg/g) e q e q e q b b b Wine tannin (mg/g) b b b q e q e q e Skin tannin (mg/g) Seed tannin (mg/g) Shiraz grapes Renata Ristic University of Adelaide
12 Seed tannin extraction is more variable than skin tannin Cabernet Sauvignon 0.02 PC2 Scores SEED SEED SEED SEED SEED SEED SEED SEED 0.01 SEED SEED SEED 0 SEED SEED SEED SEED SKIN SKINSKIN SKIN SKIN SEED SEED SEED SEED SEED sg nm, X-expl: 89%,10% PC1 Helen Holt AWRI 12
13 The problem with Pinot phenolics Low total anthocyanins No acylated or coumarylated anthocyanins Grapes have high tannin but it s mostly seed tannin Wines have relatively low tannin and colour Tannin (g/l) Grape extracts Cab Sav Grenache Merlot Variety Pinot Noir Shiraz Tannin (g/l) Wine CAF CAS DUR GRE MER Variety NEB PIN SAN SHZ 13
14 No free anthocyanins in old winespigmented tannins dominate colour Vintage 2004 col dens hue 0.70 ma U g ( ) Del-3-G Cya-3-G Pet-3-G M3G Peo-3-G M-3-G "Vitisin A" Del-A-G anthocyanins Pet-A-G Peo-A-G M-A-G M3AG Pet-C-G Peo-C-G or Pinotin A M-C-G M3CG PP 2004 pigmented tannins Vintage 1954 col dens 5.50 hue 1.32 ma U g ( ) Peo-C-G or Pinotin A PPpigmented tannins
15 The physics of gravitation has Einstein s equation for general relativity
16 The chemistry of wine colour has the Dambergs-Herderich equation [PP] = 0.06 [ M3G ] [ T ] R 2 = 0.92 PP: pigmented polymers M3G: malvidin 3-glucoside T: tannins Can we compensate for low anthocyanins in Pinot noir by boosting soluble tannin to promote pigmented tannin formation?
17 Tannin and pigment correlate with quality TANNIN (g/l) PIGMENT (AU) PRICE (USD) PRICE (USD)
18 Tannin, pigment and quality: wine show performance cluster analysis 4 Scores S B The judges got it wrong! PC-2 (35%) High pigment Low tannin S B N B N N B B B N N N B S S S N High pigment B G S High tannin B N B N GB N N N N B B B B S B G B N PC-1 (64%) N N N S SN B N Low pigment, Low tannin B N B B High Ntannin Low pigment Class 18, 2 yo Pinot, Tasmanian Wineshow Best wines have an ideal combination of tannin and pigment
19 Tannin and pigment analysis correlates with sensory PC-2 (21%) whole bunch Bi-plot 116 Herbal Stalky/sappy Tannin taste Spice Hue Tannin analysis 321 Fruit flavour Total phenolics Cherry destemmed Visual Colour, Plum Pigmented tannin Colour density Pinot Massif Total Pigment PC-1 (35%)
20 Things to do list (WineTasmania Field Day 2008) Benchmarking of phenolics in Pinot Noir grapes Benchmarking of phenolics in Pinot noir wines Examine maceration options during ferment Examine effects of additions during ferment Examine yeast/bacteria options during ferment Initially on a small-lot winemaking scale, but if promising expand to production level 20
21 Not for the faint hearted Hot maceration Rapid extraction of tannin and anthocyanins Cold fermentation Preserve fermentation volatiles Less cap work required 21
22 Optimising phenolic development during red wine maceration Need to synchronise extraction of anthocyanin extraction of tannin (skin, seeds and stems?) availability of active yeast metabolites promotion of anthocyanin/tannin reactions to form stable pigments
23 Control points during vinification (The fruit) Crushing destemming, whole bunches, whole berries Preferment maceration cold soak, enzyme, heating Co-fermentation with skins, other varieties Juice runoff Yeast type grape aroma release, fermentation esters, mouthfeel, colour/tannin stabilisation Cap management, temperature, fermentation time Seed removal or addition Post ferment maceration Malolactic ferment Oak
24 Small-lot winemaking The sorting table Fermentation monitoring The weighbridge The tank farm The crusher/destemmer The Bodum fermenter/filter/press Tall, narrow settling tank
25 10 kg submeged cap fermentation bucket
26 The flyscreen fermenter (200g)
27 Measurement of wine tannin and anthocyanin HPLC can be used to separate and quantify individual phenolic compounds BUT.complex, expensive and slow
28 The Somers measurements: Colour expression of free anthocyanins is sensitive to ph and SO 2 Malvidin-3-glucoside ph<1.5 ph 3.7 ph 3.7+SO 2 Polymerised pigments (PP) are less affected by ph and SO 2 Differential sensitivity of anthocyanins and PP forms the basis of the Somers measurements Use a combination of absorbance at 420 and 520nm with and without SO 2 and low ph treatment to estimate anthocyanins and PP plus colour density and hue 28
29 Bob s modified Somers analysis Samples are diluted in a wine-like buffer with with acetaldehyde added with high SO 2 added (read at 420 and 520 nm) provides control of ph, ethanol to minimise their effects on colour samples can be read in 10 mm cuvettes instead of 1 mm can be adapted for use in a plate reader spectrophotometer Samples diluted in 1M HCl (read at 280 and 520 nm)
30 Tannin and anthocyanins have distinct UV spectra Malvidin-Coumaryl-Glucoside Malvidin-3-Glucoside Tannin Variables M3G M3GCoum tannin
31 The spectral fingerprint of tannin is buried in wine UV spectra Chemometrics: a statistical method to extract chemical information from data systems Variables d1shz1 d1shz2 d1shz3 d1shz4 d1shz5 d1shz6 d1shz7 d1shz8 d1shz9 d1shz10 d1shz11 d1shz12 d1shz13 d1shz14 d1shz15 d1shz16 d1shz17 d1sh
32 Multiple Linear Regression (MLR) calibration to calculate tannin with 5 UV wavelengths Predicted Y Elements: Slope: Offset: Correlation: RMSEP: SEP: Bias: Spectral Tannin e-06 TCAS TCAS SHZ SHZ CAS CAS CAS SHZ SHZ SHZ SHZ SHZ SHZ SHZ SHZ CAS SHZ CAS DUR SHZ SHZ DUR CAS CASHZ SHZ SHZ SHZ CAS SHZ DUR CAS SHZ CAS DUR SHZ CAS SHZ SHZ CAS CAS SHZ CAS CAS SHZ SHZ CAS SHZ SHZ MER CAS CAS CAS SHZ SHZ SHZ SHZ CASCAS SHZCAS SHZ CAS MER CAS CAS SHZCAS CAS SHZ CAS SHZ CAS SHZ SHZ MER MER SHZ SHZ CAS SHZ SHZ CAS CAS SHZ SHZ SHZ MER MER SHZ SHZ SHZ MER CAS SHZ CASMER SHZ SHZSHZ SHZ SHZ SHZ MER CAS SHZ MER MER CAS CAS MER CAS SHZ SHZ CAS MER CAS SHZ PIN SHZ DUR PIN SHZ MER MER TCAS TCAS SHZ MER SHZ DUR CAS PIN CAS PIN MER TCAS TCAS TCAS TCAS PIN SHZ MER MER SHZ MER MER PIN TCAS TCAS TCAS MER TCAS TCAS TCAS PIN CAS PIN PIN PIN PIN PIN MER PIN PIN Methyl cellulose reference Tannin Measured Y tannin raw 250,, Y-var: tannin R 2 = 0.92 Same standard error as reference method: 0.2 g/l
33 Simplified wine analysis Modified Somers and tannin analysis Colour density Hue Anthocyanin* Total pigment* Total phenolics* Pigmented tannin* Total tannin* *can be analysed with the AWRI Tannin Portal
34 Correlation matrix Somers & HPLC Col_Den Anth_Som A520_SO2 Phen M3G PP Col. Density 1.00 Antho_Somers A520 in SO Phenolics M3G_HPLC PP_HPLC Tannin_HPLC KISS Somers total anthocyanins can replace HPLC anthocyanins Somers colour density or A 520 in high SO 2 can replace HPLC pigmented polymers (and new rapid tannin methods can replace HPLC tannin?) 34
35 Phenolics 101 Tannin Total tannin i.e. pigmented and non-pigmented Increases slowly while wine is on skins (extraction needs alcohol and heat) Skin tannin is more readily extracted than seed tannin Decreases slightly with aging Total phenolics Anything that absorbs UV at 280 nm All forms of tannin, anthocyanins, phenolic acids, flavonols etc Total Pigment Free anthocyanin and pigmented tannin Increases while wine is on skins then decreases gradually with age Free anthocyanin Increases quickly while on skins (freely soluble) Decreases quickly off skins (after 5 years all consumed) Pigmented tannin - pigmented polymers, non-bleachable pigment Formation starts during fermentation Gradual increase after wines taken off skins and during maturation Formation promoted by yeast metabolites Formation promoted by micro-oxidation and barrel maturation
36 Experimental winemaking treatments Control Cold macerate 4 days at 4 C Extended post-ferment maceration (45 days) 20% juice runoff before fermentation 20% juice runoff, returned in 2 stages near end of ferment Stems added back Oak powder added all innoculated with RC212 submerged cap ferments, 28 C
37 Tannin can be manipulated during fermentation 2.5 TANNIN 70 TOTAL PHENOLICS TANNIN (g/l) TOTAL PHENOLICS (AU) ExtMac45 cold_macer control oak TREATMENT run_return runoff stems 50 ExtMac45 cold_macer control oak TREATMENT run_return runoff stems Not equivalent to tannin!!
38 An example of rapid colour stabilisation 600 ANTHOCYANIN 4 PIGMENTED TANNIN ANTHOCYANIN (mg/l) PIGMENTED TANNIN (AU) more stabilised colour!! 100 ExtMac45 cold_macer control oak TREATMENT run_return runoff stems 0 ExtMac45 cold_macer control oak TREATMENT run_return runoff stems
39 Cluster analysis (PCA)- all samples Scores 3 2 stems runoff Phenolics Tannin stems 1 ExtMac45 ExtMac45 run_return stems run_return runoff cold_macer PC-2 (21%) ExtMac45Tannin Hue Pigmented tannin control oak control run_return control runoff cold_macer cold_macer PC-1 (68%) oak oak Colour density Anthocyanin Total pigment cold_macer control ExtMac45 oak run_return runoff stems
40 Putting it in commercial context The Pinot Shop...Avancé is a very charming pinot. The fruit has a sweet suppleness. It is quite delicate but there s nice liveliness and freshness with florals and red berries.. $29.50 Tannin 1.31 g/l Total Pigment 8.11 AU Non-bleachable pigment 1.14 AU % non-bleachable 10.0 The Pinot Shop Reveur ( dreamer ) Pinot Noir is a pure but weighty wine with distinct charry-cherry characters. That s from some good oak wrapped around vintage 2008 s ripe Coal Valley fruit. The tannins are round and chewy I suspect that the ripe tannins will smooth out into silky, luscious pinot.. $46 Tannin 1.84 g/l Total Pigment AU Non-bleachable pigment 1.56 AU % non-bleachable 13.7
41 Moving it to the next level RC 212 control EC1118 Bayanus: AWRI 1176, AWRI 1375 Hybrids: AWRI Fusion, AWRI 1503 TD Carbonic maceration White skins Cold soak, then wild ferment Cold soak, wild primary, delayed wild malo Co-fermentation with Pinot Gris or GT Stalks Transfusion (run off juice and return near end of ferment) ICCS Workshop 2- Taming the Pinot noir terroir with Nick Glaetzer and Jenny Bellon
42 Taming tannin 1.7 TANNIN (g/l) _Fusion 2TD Amarone Carbonic Chard+GT Cold soak Delayed cold GT Pinot Gris Stalks Transfusion X_Control White skins or stalks added had the highest tannin Alternative yeasts lower than RC212 Transfusion higher than control
43 Taming pigmented tannin PIGMENTED TANNIN (AU) _Fusion 2TD Amarone Carbonic Chard+GT Cold soak Delayed cold GT Pinot Gris Stalks Transfusion X_Control Cold soak/delayed malo had highest but it was an unusual plummy colour (Hue_SO 2!!) Elevated with most alternative yeasts except for 1176 Elevated when white skins and stalks added Elevated with transfusion
44 The ratio of pigmented tannin and total tannin PigTannin/Tannin _Fusion 2TD Amarone Carbonic Chard+GT Cold soak Delayed cold GT Pinot Gris Stalks Transfusion X_Control Highest were alternative yeasts and cold soak/delayed malo Does it affect mouth-feel?
45 Colour density Corrected for SO 2 High colour density = high visual colour 8 COLOUR DENSITY (AU) _Fusion 2TD Amarone Carbonic Chard+GT Cold soak Delayed cold GT Pinot Gris Stalks Transfusion X_Control Cold soak/delayed malo had highest Next were white skins and stalks!! Most alternative yeast higher than control but not 1176
46 Anna Carew: microwave maceration and yeast effects TIAR research development extension education training
47 Yeast strain and total tannin 0.6 b Total tannin (gm/l) a* a c c EC1118 RC212 AWRI1176 w-ec1118 T.d-EC1118 Yeast treatment *Means with the same letter are not significantly different at the p 0.05 level according to Tukey s Test. TIAR research development extension education training
48 Microwave maceration TIA/AWRI have conducted 30+ replicated trials of microwave maceration for since Compared with control fermented on skins for 8 days, microwave wines showed more rapid and effective extraction of phenolics. Thus far, applied to Pinot noir and Shiraz musts, at laboratory scale (1kg). 1. Intermittent microwave & stirring 2. Monitor for peak temperature (70 C) 3. Hold time in 70 C waterbath 4. Cool to ~24 C, inoculate with ADY for AF
49 Microwave maceration decreases total yeast count more effectively than SO 2 : Pasteurisation effect means no need for SO 2 addition at crushing SO 2 Microwave C&D(no SO 2 )...and Botrytis laccase!! 1.5hr Post-mac 3.0hr Post-mac
50 Options for winemakers Rapid extraction by microwave of phenolics offers two options: EARLY PRESS-OFF: Press-off, cool and ferment juice ( mpr ) FERMENT ON SKINS: Cool the must and conduct alcoholic ferment on skins ( msk )
51 Wine phenolics On skins Early press off Anthocyanins (mg/l) Pigmented tannin (AU) Total tannin (g/l) CTL MSK MPR 292 a 412 b 272 a 0.31 a 0.50 b 0.45 b 0.25 a 0.72 b 0.27 a Different letter denotes significant among treatments within trial (P<0.05)
52 Phenolic Release by Microwave control heat microwave % cells with phenolic material 40% 35% 30% 25% 20% 15% 10% 5% 0% a a b control heat microwave TIAR research development extension education training b Histology: Dr Warwick Gill, TIA
53 Taste? control microwave Informal tasting (Godden, Wilkes, Dambergs) Microwave: AROMA more (red & dark) fruit, pinotlike MOUTH more palate weight, nice length, soft and mouth-coating, not hard tannin Control: AROMA and MOUTH no faults but pissy
54 Aroma Differences by GCMS Ethyl acetate ctl & msk ~1.4 mpr 2.3 Ethyl octanoate ctl & msk ~1.7 mpr 3.7
55 Commercial tunnel microwave Semi-commercial scale - 15 kw pentagonal microwave unit, ~200kg/hr (with Dr Kai Knoerzer, CSIRO Animal, Food and Health Sciences, VIC)
56 Angela Sparrow: The contribution of grape compartments to wine tannin and colour TIAR research development extension education training
57 Increasing grape pulp reduces wine tannin Double bunch part: wine tannin Tannin (g/l) berries 2Xskins 2Xseeds 2Xpulp green stalks Bunch part mature stalks
58 Removing grape pulp increases wine tannin Missing bunch part: wine tannin Tannin (g/l) whole berries no skins no seeds no pulp skin only pulp only seeds only juice only Bunch part
59 Does seed tannin produce stable colour? (a) 15 % Non-bleachable pigment LSD 0.05 (b) 2 Non-bleachable pigment (AU) LSD Bottling 6 months 12 months seeded seedless 0 Bottling 6 months 12 months PN Control PN & Ferm seed PN & GS tannin
60 Selective skin maceration to enhance extraction of skin tannin LSD Tannin g/l Non-bleachable pigment (AU) LSD Colour Density (AU) LSD Bottling 6 months 0.0 Bottling 6 months 0.0 Bottling 6 months cut cap D1 cap not cut cut cap D3 cut cap D5
61 Adding fresh white pomace
62 The winemaking control points Extended maceration wines have high tannin and a high degree of colour stabilisation Cold maceration favours colour extraction Juice runoff (saignee) results in higher tannin and colour but favours colour more Running off juice and returning it later during ferment (transfusion) increases tannin and stable colour The selection of yeast strain has dramatic effects on tannin and colour Boosting tannin with a non-pigmented source (eg stems, white skins) can also increase colour but is it stable long-term? Is there a place for microwave maceration? Can this create a new winemaking paradigm? Do we really want seed tannin? How can we enhance skin tannin extraction?
63 THE FINAL WORDS Pinot noir tannin, total colour and colour stabilisation can be strongly influenced by maceration/vinification methods, including the choice of yeast For a given parcel of grapes, tannin in particular, can be doubled through manipulating vinification : Bob Dambergs 13 March 2009 turning on the tap
64 Acknowledgments The ICIP Consortium AWRI, Croplands, Flextank, Tamar Ridge, TIA, Wine Tasmania Industry contributors Clover Hill, Frogmore Creek, Jansz, Josef Chromy, Meadowbank, Moorilla, Pooley, Tamar Ridge, Tolpuddle, Winemaking Tasmania The funding bodies
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