micro-oxygenation and its impact on polyphenols and … · micro-oxygenation and its impact on...

November 18th, 2014ASVO Seminar Inputs to Outputs; is less, more?

Micro-oxygenation and its impact on polyphenols and sensory characteristics

of red wines

Dominik Durner1,2, Patrick Nickolaus1, and Hai-Linh Trieu1

1 Dienstleistungszentrum Ländlicher Raum (DLR) Rheinpfalz, Department of Viticulture & Enology, Breitenweg 71, 67435 Neustadt an der Weinstraße, Germany

2 Hochschule Kaiserslautern, Department of Applied Logistics and Polymer Sciences ,Carl-Schurz-Str. 10-16, 66953 Pirmasens, Germany


Viticulture in Germany

Neustadt

Source: Weingut Stephan Fischer

Source: Pfalzwein e.V.

Source: DWI


DLR Rheinpfalz - State Research and Education Center for Viticulture and Horticulture

© DLR Rheinpfalz © DLR Rheinpfalz

© DLR Rheinpfalz© DLR Rheinpfalz


Which one is your money on?

No Mox Post-MLF Mox(5/12wk)

Pre-MLF Mox(20/3wk) No Mox Post-MLF Mox

(5/12wk)

2013 Pinot noir (pre bottling)--------20 % Botrytis---------

Picture by Marlene Neser


0%

20%

40%

60%

80%

100%

120%

0 1 2 3 4 5 6 7 8 9

Per

cent

age

of m

ax. c

onet

ratio

n

Time (days)

Alcohol (89.4 g/L)Anthocyanins (182 mg/L)Tannins (103 mg/L)

Evolution of anthocyanins and tannins during fermentation

2003 Pinot noir; 50 hL fermentation on skins (28°C/8 d); punch downs (4-12-4 per day)

Markus Löchner (2004)


0

2

4

6

8

10

12

14

0

50

100

150

200

250

300

Pre AF AF(3 d)

Post AF(10 d)

Pre MLF(2 m)

Post MLF(3 m)

Post MOX(6 m)

SO2(7 m)

Bottling(9 m)

3 m post-b(12 m)

9 m post-b(18 m)

Col

or D

ensi

ty(A

U)

Ant

hocy

anin

s(m

g/L)

Anthocyanins (mg/L)

Colour Density (AU)

2012 Pinot noir; thermovinification (85°C/2 min); 3 hL juice fermentation (20°C/10 d); MLF (18°C/35 d); micro-ox (10 mg/L/m/12 wk); 90/35 mg/L total/free SO2; bottle storage (15°C/9 m)

Evolution of colour from must to bottle

Julia Willeke (2014)


Reasons for the loss of anthocyanins

1. Polymerisation/derivatisation reactions nice red pigments precipitation of polym. pigments

2. PPO catalysed oxidationof anthocyanins brownpigments or colourless adducts

3. Adsorption to yeast cells orfining agents loss in colour

4. ß-Glucosidase-catalyzedcleavage low solubility loss in colour

0

10

20

30

40

50

60

Rem

aini

ngan

thoc

yani

nsat

bottl

ing

(%)

DV 10IOC

Pinot noir from vintages 2013, 2010, and 1979

Picture by Patrick Nickolaus Julia Willeke (2014)


Cons

• Risk of over-dosing

• High SO2 demand

• Oxidised aroma

• Dry tannins

• Stuck MLF

• Fast aging

• Browning

• No effect

Pros

• Stable colour pigments

• Early drinking maturity

• No reductive flavour

• Higher fruit intensity

• Less green tannins

• Balanced tannins

• Microbial safety

• Less CO2

Micro-oxygenation


Barrels HDPE-tanks Synthetic cylindricals

New 225 L barrel:3-4 mg O2/L/month1

Used 225 L barrel:1-2 mg O2/L/month1

5000 L HDPE-tank:0.4 mg O2/L/month2

300 L HDPE-tank:2.1 mg O2/L/month2

Low OTR:0.01 mg O2/L/month3

High OTR:0.7 mg O2/L/month3

1 from Vivas (1997); 2 Flecknoe-Brown (2005); 3 product information Nomacorc Select Series

Passive supply of oxygen


Micro-oxunit

O2

0.1 bis 180 mg O2/L/month

Active supply of oxygen (micro-ox)


• Exact und continuous dosage of oxygen• Production of small bubbles Slow ascent velocity of bubbles

• No sticking bubbles to diffuser Growing bubbles

• Quick dissolution of oxygen in the wine no accumulation of oxygen

• Good distribution of oxygen in the tank

Technical prerequisites

0 20 40 60 80 100

3 m

2 m

1 m

0.1 m

Dissolved oxygen (µg/L)

Dep

thfro

mw

ine

surfa

ceDiffuser at 3.5 m; n=3


Semiquinone

•

Fe2+

Fe3+

O2

HO2•

Fe2+ Fe3+

H2O2

adapted from Danilewicz (2003) Am J Enol Vitic 54: 73-85.

OH• Fe3+

Fe2+

Formation ofoxygen radicals

Fenton-Reaction

CH3CHOAcetaldehyde

CH3CH2OHEthanol

Semiquinone

Quinone

Catechol

•

H2O

Oxygen-induced formation of acetaldehyde


O+

OGlcOH

OH

OH

OCH3

OCH3

Anthocyan (Mv)

Vitisin B (R=H) Mv-mm-Cat

GlcO

OCH3

H3CO

OH

OH

OH

OH

OH

O

O+ OH

OH

OH

+ Acetaldehyde

Acetaldehyde-induced reactions of anthocyanins

O

OH

OH

OH

OH

OH

R

+

Catechin (Cat)

+ Acetaldehyde

Hadapted from Fulcrand (1998) Phytochem 47: 1401; Timberlake et al. (1976) Am J Enol Vitic 27: 97.


2013 Pinot noirVinyard Oberderdingen (Württemberg)Harvest Hand-picked on October, 12th, 2013 Gravity 20.9°BxpH 3.2YAN 240Chaptalising 110 g/L pot. Alc.

WinemakingTemperature (AF) 28°CPunch downs 4-12-4 per day (1 min)Yeast Mycoferm Cru 05Simultanious MLF Extremo IT14Pressing after 8 daysTemperature (MLF) 20°CRacking after 36 days

Winemaking protocol


Micro-ox experiment

2013 Pinot noir

Control

0 mg/L 10 mg/L

Grape seed tannins (20 g/hL)

20 mg/L 0 mg/L 10 mg/LOxygen supply per month for 12 weeks

Each experiment was carried out in duplicate.

Height: 3 mVolume: 100 L

Temperature: 15 ºC


30

40

50

60

70

80

90

100

110

120

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Nat

hocy

anin

s(m

g/L)

Time (weeks)

Control10 mg/L/month20 mg/L/month

Anthocyanins during micro-ox:Impact of different oxygen rates

End12 wk micro-ox


4

5

6

7

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Col

or D

ensi

ty (A

U)

Time (weeks)


Colour during Micro-ox:Impact of different oxygen rates

End12 wk micro-ox


End12 wk micro-ox

0

20

40

60

80

100

4

5

6

7

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Ace

tald

ehyd

e (m

g/L)

Col

or D

ensi

ty (A

U)

Time (weeks)


Total SO2: 109 mg/L (a)

Total SO2: 81 mg/L (b)

Total SO2: 84 mg/L (b)

4 wk after Mox: 35 ppm free SO2

Acetaldehyde during Micro-ox:Impact of different oxygen rates


4

5

6

7

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Col

or D

ensi

ty (A

U)

Time (weeks)

Control10 mg/L/monthTanninTannin + 10 mg/L/month

Colour during Micro-ox:Impact of tannins and different oxygen rates

Tannin effectw/o O2

Tannin effectwith Mox

End12 wk micro-ox


End12 wk micro-ox

0

20

40

60

80

100

4

5

6

7

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Ace

tald

ehyd

e (m

g/L)

Col

or D

ensi

ty (A

U)

Time (weeks)

Control10 mg/L/monthTanninTannin + 10 mg/L/month





4 wk after Mox: 35 ppm free SO2

Acetaldehyde during Micro-ox:Impact of tannins and different oxygen rates


Vitisin B and anthocyanin-catechinoligomers after micro-ox

0

5

10

15

20

25

30

Control 10 mg/L/month 20 mg/L/month Tannin Tannin+ 10 mg/L/month

Con

cent

ratio

n (m

g/L)

Trimer

Dimer

Vitisin B


0.0

0.2

0.4

0.6

0.8

1.0

Monomer Dimer Trimer

Abs

orpt

ion

pH 1pH 3.5pH 5

Absorption at 520 nm(1 g/L pure substance in model wine)

0

1

2

3

4

5

6

Monomer Dimer Trimer

Abs

orpt

ion

AbsorptionAbs. after SO2

Absorption at 620 nm(1 g/L pure substance in model wine)

Color properties of malvidin vs. anthocyanin-catechin oligomers

εpH1= 2200 L mol-1 cm-1

εpH1= 3137 L mol-1 cm-1

εpH1= 3892 L mol-1 cm-1

Molar absorptivity at 520 nm Ethylidene bridges:Batochromic shift

at low pH

Weber et al. (2014) Food Res Int (in press)


PC

2(3

2%

)

-4

-2

0

2

4

-4 -2 0 2 4PC 1 (49 %)

No tannin addition20 g/hL tannins

Control

210mg O /L/month

220 mg O /L/month

10 mg O2/L/month

Control

Sensory impact of tannins and Micro-ox in 2013 Pinot noir 3 month after bottling

Descriptive analysisin June 2014;

n = 18 × 2; Scale: 0-10; Significance levels:

* p ≤ 0,05** p ≤ 0,01;

*** p ≤ 0,001


Pro

porti

on o

fant

hocy

anin

sin

pol

ymer

icpi

gmen

ts(m

g/g)

0

20

40

60

80

100

14 polym. phenolic fractions separated by Sephadex LH20

Incorporated anthocyanins in polym. phenolicfractions in a micro-oxed 2005 Cab. Sauv.

Weber et al. (2013) Am J Enol Vitic 64: 15

Descending molecular weight


Weber et al. (2013) Am J Enol Vitic 64: 15

0

2

4

6

8

10A

strin

genc

yin

tens

ity

fgg

efg defgcde

efgcdef cdef

bcd bc bcd

aab

a

Descriptive analysisIso-concentrations in watern = 13 × 2; Scale: 0-10;

Values sharing the same letter are not significantly different at

p < 0.05.

14 polym. phenolic fractions separated by Sephadex LH20

Descending molecular weight

Astringeny of polym. phenolic fractions in a micro-oxed 2005 Cab. Sauv.


• Micro-ox only if laccase-free [and microbial safe]

• Micro-ox as early as possible: After AF (not during MLF)

• For Pinot some 5-10 mg O2/L/month after MLF for 3 months help for colour stabilisation and lower green tannin perception[Pre-MLF equivalent: 20-30 mg O2/L/month for 4 weeks]

• Slight accumulation of acetaldehyde is OK, as long as it doesn’t cause a higher SO2 demand

• A good proportion of tannins is needed for the formation of polymeric pigments

• Polymeric pigments does not only seem to improve colour, but also to soften astringency perception

Conclusions


Patrick Nickolaus, Hai-Linh Trieu, Fabian WeberJulieta Luna Zullo, Annie-Michelle Nno-Essaa,Julia Willeke, Uli Fischer, Hans-Georg Schmarr,Anette Schormann, Sascha Wolz,Peter Winterhalter, Eva Schmalfuss

This research project is supported by the German Ministry of Economics and Technology (via AiF) and the Research Association of the German Food Industry (FEI) in cooperation with the Association of German Wine-growers (DWV) and the Association of Cooperative Wineries Baden Württemberg (BWGV). Project AiF-FV 17719 N

Industry partners for co-funding, providing equipment, and enological supplements: Nomacorc, Ever Intec, Lallemand, WZG

Thanks to

micro-oxygenation and its impact on polyphenols and … · micro-oxygenation and its impact on...

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