Dr. Christian E. BUTZKE Associate Professor of Enology ...Associate Professor of Enology Department of Food Science PURDUE UNIVERSITY [email protected] 1-765-494-6500 . ... Robert

Dr. Christian E. BUTZKEAssociate Professor of EnologyDepartment of Food Science

PURDUE [email protected] 1-765-494-6500 www.purdue.edu

IntroductionTCA History and StatisticsTCA Formation Manufacturing Process AnalysisTCA PreventionTCA Thresholds and Sub-ThresholdsCork Quality ControlTCA Analyses in Wine and CorksOxygen Permeability and Gas ExchangePersonal Outlook on TCA and Closures

IntroducciònTCA Historia e EstadísticaTCA FormaciónAnálisis Del Proceso De FabricaciónTCA PrevenciónTCA Umbrales de Detección e ReconocimientoControl de Calidad de Tapónes de CorchoTCA Análisis en Vino y Tapónes Permeabilidad del Oxígeno y Intercambio del GasPerspectiva personal en el TCA y los Tapónes


Mi 11ma Visita a Cataluña(desde 1967)

Palma de Mallorca, 1993

Robert Mondavi Robert Mondavi


1904 Lefebre et al: “Goût de Bouchon”1981 Tanner et al: TCA identified as cork taint1990 Simpson et al: TCA formation revealed1996 Butzke et al: TCA threshold variation1997 Butzke et al: SPME GS/MS-SIM for TCA1998 Butzke et al: Cork Sensory Quality Manual1998 Butzke et al: Automated SPME for TCA2000 Butzke et al: Supercritical CO2 Extraction 2002 Butzke et al: Sub-threshold TCA in wine2005 Butzke et al: ASEV “Science of Closure”

1996 Butzke et al: TCA threshold variation1997 Butzke et al: SPME GS/MS-SIM for TCA1998 Butzke et al: Cork Sensory Quality Manual1998 Butzke et al: Automated SPME for TCA2000 Butzke et al: Supercritical CO2 Extraction 2002 Butzke et al: Sub-threshold TCA in wine2005 Butzke et al: ASEV “Science of Closure”

All for € 2500!

Cl

Cl

Cl

OCH3

2,4,6-TriChloroAnisole

= Musty Taint

AWRI: 80+% of tainted wines

Producción De Vino

Gallons x 1000

010002000300040005000600070008000

1990 2000 2005

El MundoEuropaEspañaEEUU

Producción De Vino

Gallons x 1000

4500

5000

5500

6000

6500

7000

7500

8000

1990 2000 2005

El MundoEuropa

-8%

-18%

Producción De Vino

%

0102030405060708090

100

1981 2004

Tapónes decorchoROTE

-17%

¿Dirección De la Industria Del Corcho 1981-2005?

Stopper Imports 800 million/year

Import Value of Corks $100 million/year

Cork Taint Rate 1 - 8%

Tainted Wine $100 - 800 million/year

Total Cost $200 - 900 million/year

EEUU Cork Statistics


Cork Taint Formation

Penta/Tetra-Chlorophenol

Lignin Phenol

Chloroanisole

ChloroanisoleChlorophenol

MethylationChlorination

Lignin Phenol ChloroanisoleAnisole

Methylation Chlorination

Methylation


Tri-Bromophenol

Lignin Phenol

Bromoanisole

BromoanisoleBromophenol

MethylationBromination

Lignin Phenol BromoanisoleAnisole

Methylation Bromination

Methylation


Tri-Chatophenol

Lignin Phenol

Chatoanisole

ChatoanisoleChatophenol

MethylationChatonnetization

Lignin Phenol ChatoanisoleAnisole

Methylation Chatonnetization

Methylation


Tri-Iodophenol

Lignin Phenol

Iodoanisole

IodoanisoleIodophenol

MethylationIodination

Lignin Phenol IodoanisoleAnisole

Methylation Iodination

Methylation

TCA Formation

Always 3 circumstances:

• Phenolic Material Wood

• Chlorination Chlorine

• Methylation Mold


Soil exposure

Storage Post-Harvest

Storage Post-Boiling

Punching out

Moldy strips

Post-Processing Storage

Post-Processing Storage

(Post-) ShipmentStorage

TCA Formation Critical Processing Points

Air pollution/herbicide use in forestStorage in forestMold growth after boilingCross-contamination during punchingBleaching (hypochlorite still in use?!)Rinsing (Clorine residues, water quality)Cross-contamination during/after dryingMoisture level post-processingShipping/storage conditions



AutoclavingSulfur DioxideEthylen DioxidePlastic CoatingsUV LightGamma RadiationMicrowavesBeta RadiationEnzymesAntibioticsSymbios (?)Dream Taste (“Ionic Filteration”?)

Anti-TCA Techniques1995-2005

Penetration 4 cm?

Vegetative cells, D12?

Mold spores?

Methylation enzymes?

TCA molecule?

Accelerated Electron Beam(Beta Radiation)

Accelerated Electron Beam(Beta Radiation)

(Post-) ShipmentStorage

en California

TCA in the Winery?

TCA in the Winery?

TCA in the Winery?

TCA in the Winery?

TCA in the Winery

TCA in the Winery?

TCA in the Winery?

TCA in the Winery?

TCA in the Winery?

Rehydration

Ideal cork moisture: 5.5 - 8%• > 8% mold growth• < 5.5% reduced recuperation

Raw cork shipped < 5.5%• Rehydration: ozonated (O3) water

¡Una Vieja Advertencia!


Comparison of 2,4,6-TCA Sensitivity inExperienced vs Inexperienced Subjects

0

1

2

3

4

5

6

71.

0

2.5

5.0

10 25 50 75 100

250

500

750

1000

2500

5000

7500

1000

0

2500

0

Concentration (ng/L)

Freq

uenc

y

2 2 2

6

3 3

1 1

3

ExperiencedPanelists (n=23)

1

2

1

4

2

1

2

1 1

InexperiencedPanelists (n=15)

TCA Sensory Thresholds Variation: 1 - 25,000 ng/L

Sub-threshold TCA

Detection Threshold

≈ ½ Recognition Threshold

=>Reduced varietal aroma noticeable at0.5 ng/L for experienced tasters

Wine Magazines & Cork Taint

Wine Spectator March 2004


Corks Problems

Taint

Oxidation

Leaking/Seepage

Irremovable Corks

Pushed-out Corks

• Sampling Procedures• Case Study• Taste Panel Evaluation• Taste Panel Training

UC Davis CorkSensory QC Manual

Butzke & Suprenant

• Testing every wine stopper economically impossible

• Method based on testing 50 corks in 10,000

• Testing method measures ‘releasable TCA‘

• 50 corks are soaked for 24 h in white wine with 10% alc

• Chemical TCA analysis of the group soak as low as 1 ng/L

• Group soaks reflect the average gained by individual soak

Cork QC Manual: 7 years later

“Amorim has introduced

sophisticated QC procedures”



Butzke & EbelerCork Research

SensorySensoryQCQC

SamplingSamplingProceduresProcedures

SPME/GCSPME/GC--MSMSTaint AnalysisTaint Analysis

SFE/ELISASFE/ELISATCA AnalysisTCA Analysis

Solid Phase Micro Extraction

SPME/GC-MSextracted ionchromatogram with stable isotope 2H5-TCA*as internal standard.

SPME

Limit of detection: 0.5 ng/L

*Courtesy ofDr. Mark Sefton, AWRI

Pressure(atm)

350 S O SUPERCRITICAL 300 L I FLUID 250 D LIQUID

200

150

50 Sublimation GAS Line 0

-60 -40 -20 0 20 40 60 80 (°C)

Pc=74.8atm Tc = 31.1°C

Phase (pressure-temperature) diagram for CO2 : CP= critical point, TP= triple point, Pc= critical pressure, Tc= critical temperature.

Supercritical Fluid Extraction

Super-criticalCO2

Goal• Rapid, sensitive, 'solventless’

method to quantitativelyextract TCA from corks

Application• Study transfer of TCA

from cork to wine• Determine TCA concentration

gradients within a cork• Determine source of TCA in

tainted corks and wines• Eliminate TCA from cork

Oven with extraction vessel (cell) Solvent Bottle Nitrogen

Solvent pump pump Restrictor SFE CO2 Cryogenic Trap Cooling CO2 Collection vial


Method Development• CO2 density of 0.837 g/mL (250 atm, 50°C)

provides optimal extraction efficiency

Spiked Samples• 98% of spiked recovery of ground cork• Limit of Detection: 5 ng• Limit of Quantitation: 10 ng

Analysis of 'Corked' Retail Wines• 200 to 2100 ng TCA per gram of cork



Oxygen Permeability of Corks

00.10.20.30.40.50.60.70.80.9

1

0.001 0.01 0.1 1 10

Permeability (CC O2 /Closure/Day)

Frac

tion

of

Clos

ures

(c

umul

ativ

e)

Synthetic Corks

Bark Corks

© Richard Gibson, Scorpex.net

Closure Testing - Mocon

© Richard Gibson, Scorpex.net

Oxygen Permeability of Corks

1000-fold variation in natural corks!

⇒ unpredictable “breathing” of winethrough the cork

Corks and Bottle Aging

New 60 Years25 Years3 Years

¡ Sideways !

The Physics of Wine Bottle Closures

Professor Roger BoultonDepartment of Viticulture and Enology

University of CaliforniaDavis, CA 95616-8749

Presented atAmerican Society for Enology and Viticulture

56th Annual Meeting, Seattle, WA

ASEV Science of Closures Seminar24th June 2005

Permeation– Pressure driven

Diffusion– Concentration driven

Fick’s Lawdn/dt = Deff[Ci –C0] / L

Henry’s LawKH = [C]i / Pi

Darcy’s LawdV/dt = k A [Pi-P0] / L / μ

2 Fenómenos

Permeation– Pressure driven

Henry’s LawKH = [C]i / Pi

Darcy’s LawdV/dt = k A [Pi-P0] / L / μ

Diffusion– Concentration driven

Fick’s Lawdn/dt = Deff[Ci –C0] / L

Permeation– Due to Pressure Difference, Pi – Po

– Headspace P exponential with Temperature– Independent of Concentration Gradient

Diffusion– Due to Concentration Difference, [Ci] – [Co] – Affected weakly by Temperature– Independent of Pressure Gradients

Permeation vs Diffusion

Darcy’s LawPermeation

k A ΔPµ L

k PermeabilityΔP Pressure DifferenceA Flow AreaL Depth of Mediumµ Viscosity

Fick’s LawDiffusion

- D (Ci – Co)L

J Molecular FluxD Molecular DiffusivityCi Inside ConcentrationCo Outside ConcentrationL Diffusion length

Permeation vs Diffusion

J = q =

Flow Through Porous Media(e.g. Cork)

• Gas permeability of the cork

• Layers of alternative media

• Variability in porosity* from cork to cork(*fraction of volume which is occupied by gas)

• Distribution of pore sizes

• Gas viscosity is 1/100th that of water

Important Factors:

Oxygen Radical Reactions

Vino blanco

Vino rojo

O2 +

O2 +

SO2 Loss: Natural Cork

SO2 Loss: Synthetic Cork

SO2 Loss: Screw Cap

Closure Evaluations

Oxygen Radical Reactions≠ Diffusion Loss

Nota bene:

5 .0

10 .0

15 .0

20 .0

25 .0

30 .0

35 .0

0 4 8 12 16 20 24

T im e o f D ay (H o urs )

Win

e Te

mpe

ratu

re (C

)

0 .70

0 .90

1 .10

1 .30

1 .50

1 .70

Bot

tle P

ress

ure

(Atm

)

B o ttle P ressure

W ineT em p era ture

3 m L s

4 m L s

5 m L s

6 m L s

Effect of Headspace Volume on Bottle Pressure

(and Permeation)

WineTemperature

Container Shipment-American B.D.216400 Combined (shipped 8/7/00 - arrived 8/17/00)

6065707580859095

100

-08-

07

-08-

07

-08-

08

-08-

08

-08-

09

-08-

09

-08-

09

-08-

10

-08-

10

-08-

11

-08-

11

-08-

12

-08-

12

-08-

13

-08-

13

-08-

14

-08-

14

-08-

14

-08-

15

-08-

15

-08-

16

-08-

16

-08-

17

-08-

17

Date

Top*

Tail*

Nose*

Temperature Fluctuationduring Wine Shipments

Temperature (°F)

* container position

Bottle Aging Reactions

O2 uptake:

Browning:

EC formation:

SO2 decline (W):

3.8ºC

7.8ºC

37ºC

(Ribereau-Gayon 1933)

(Berg & Akiyoshi 1956)

(Ough 1985)

Doubling of Rate

(CEB 2001)

16ºC


¿¿El El ExtremoExtremo de de CorchosCorchos??

$150 per bottle!Alcoa

Vino-Lok

AlcanStelvin


¿Un Futuro para los Corchos?


TCA Filter

Derived from ELISA-TCA test

Monoclonal antibodies for TCA

Polymer-embedded antibodies

Filter for “corked” bottle of wine

=> TCA removal on demand

¿Un Futuro para los Corchos?

Prof. Susan EbelerProf. Roger BoultonProf. Tom YoungMr. Adam SuprenantMr. TJ EvansMs. Marisa Taylor-HuffakerMr. Michael MoyerMr. Gordon BurnsCalifornia Enological Research Association

Documents

Dr. Christian E. BUTZKE Associate Professor of Enology ...Associate Professor of Enology Department of Food Science PURDUE UNIVERSITY [email protected] 1-765-494-6500 . ... Robert