Unesco conference “Vine, wine, food and health” (Heraklion, Crete, 2018). http://wineculture.chemistry.uoc.gr/wordpress/

1. An update on wine ageing and sulfonations
Panagiotis Arapitsas, Graziano Guella & Fulvio Mattivi

3. SO2

4. BackgroundAn update on wine ageing and sulfonations
ECAT-SO3H
epicatechin-SO3H
PROC-B2-SO3H
procyanidin B2-SO3H
O
OH
OH
OH
OH
OH
S
O
O
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
S
O
O
OH

5. An update on wine ageing and sulfonations
OH
N
H
S
O
O
OH
O
OH
N
H
OH
S
O
O
OH
TOL-SO3H
tryptophol-SO3H
ILA-SO3H
indole-lactic acid-SO3H
Background

6. AimAn update on wine ageing and sulfonations
OH
NH2
N
H
O
O
NH2
OH
O
OH
N
H
OH
O
OH
N
H
OH
N
H
TOL
N
OH
O
OH
N
H
O
NH O
N
H
OH
NH2
shikimic
OH
OH
OH
OOH
OOH
NH2
O
NH2
O
O
N
H
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
O
NH
N
H
O
O
O
NH2
N
H
O
epicatechin
procyanidin B2
phenylalanine
tryptophan melatoninserotonin
tryptophan ethyl ester
N-acetyl-tryptophan ethyl ester
kynurenic
anthranillic
kynurenine
indole-3-pyruvic acid
indole-3-lactic acid
indole-3-acetic acid
tryptophol
2-aminoacetophenone
OOH
NH2
anthranillic

7. An update on wine ageing and sulfonations
O
OH
N
H
OH
O
OH
N
H
OH
N
H
TOL
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
epicatechin
procyanidin B2 indole-3-lactic acid
indole-3-acetic acid
tryptophol
ECAT-SO3H
epicatechin-SO3H
PROC-B2-SO3H
procyanidin B2-SO3H
O
OH
OH
OH
OH
OH
S
O
O
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
S
O
O
OH
OH
N
H
S
O
O
OH
O
OH
N
H
OH
S
O
O
OH
TOL-SO3H
(tryptophol-SO3H)
ILA-SO3H
(indole-lactic acid-SO3H)
O
OH
N
H S O
O OH
IAA-SO3H
(indole-acetic acid-SO3H)
Aim

8. 1 11
2
3
4
5 6
7
8
9
1 10
2
3
4
5 6
7
8
9
1 10
An update on wine ageing and sulfonations Methods, synthesis
O
OH
N
H
OH
O
OH
N
H
OH
N
H
TOL
indole-3-lactic acid
indole-3-acetic acid
tryptophol
OH
N
H
S
O
O
OH
O
OH
N
H
OH
S
O
O
OH
TOL-SO3H
(tryptophol-SO3H)
ILA-SO3H
(indole-lactic acid-SO3H)
O
OH
N
H S O
O OH
IAA-SO3H
(indole-acetic acid-SO3H)
SO2 in water
indole in ethanol
+ 3 mol SO2
rt, 2 days
+ 18 mol SO2
rt, 14 days
+ 12 mol SO2
rt, 6 days

9. MethodAn update on wine ageing and sulfonations
Metabolites
ILA indole 3-lactic acid
ILA-GLU indole 3-lactic acid glucoside
ILA-SO3H indole 3-lactic acid 2-sulfonate
IAA indole 3-acetic acid
IAA-ASP indole 3-acetic acid aspartic acid
IAA-SO3H indole 3-acetic acid 2-sulfonate
IPA indole 3-pyruvic acid
ICA indole 3-carboxaldehyde
2AAP 2-aminoacetophenone
TRP tryptophan
N-TRP-EE N-acetyl-tryptophan ethyl ester
TRP-EE tryptophan ethyl ester
MEL melatonine
SER serotonine
N-SER N-acetyl serotonine
KYNA kynurenic acid
KYN kynurenine
TOL tryptophol
TOL-SO3H tryptophol sulfonate
TYR tyrosine
TYR-EE tyrosine-ethyl ester
N-TYR-EE N-acetyl-tyrosine-ethyl ester
TYL tyrosol
PHE phenylalanine
ABA abscisic acid
ABA-GLU abscisic acid glucoside
CAT catechin
ECAT epicatechin
PROC-B1 procyanidin B1
PROC-B2 procyanidin B2
ECAT-SO3H epicatechin 4-sulfonate
PROC-B2-
SO3H procyanidin B2 -sulfonate
O
NH2
N
H
OH
O
OH
OH
OH
OH
OH
O
NH2
OH
O
NH2
OH
OH
O
OH
O OH

10. winesAn update on wine ageing and sulfonations
195 wines
vintage: 1986-2016
93 white wines
35 Chardonnay
32 Pinot gris
24 Verdicchio
…
(2001-2016)
37 sparking wines
Chardonnay, Pinot noir
White, Rosé, Riserva
5 rosé still wines 60 red wines
18 Sagrantino
13 Tannat
12 Sangiovese
11 Amarone
…
(1986-2015)

11. An update on wine ageing and sulfonations
red
Results, flavanols
sparkling
white
sparkling
white
red
ECAT-SO3H
epicatechin-SO3H
PROC-B2-SO3H
procyanidin B2-SO3H
O
OH
OH
OH
OH
OH
S
O
O
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
S
O
O
OH

12. Results, flavanolsAn update on wine ageing and sulfonations
Amarone Tannat Sagrantino
6 8 10 14 16 19 24 26 31 5 6 7 14139
Age
10
Age
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
epicatechin
procyanidin B2
136 7 11 12
Age

13. Results, flavanolsAn update on wine ageing and sulfonations
ECAT-SO3H
epicatechin-SO3H
PROC-B2-SO3H
procyanidin B2-SO3H
O
OH
OH
OH
OH
OH
S
O
O
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
S
O
O
OH
6 8 10 14 16 19 24 26 31 5 6 7 14139
Age
10
Age
136 7 11 12
Age
Amarone Tannat Sagrantino

14. Results, flavanolsAn update on wine ageing and sulfonations
Amarone Tannat Sagrantino
% PROC-B2
% PROC-B2-SO3H
% ECAT
% ECAT-SO3H

15. An update on wine ageing and sulfonations Results, indoles
O
OH
N
H S O
O OH
OH
N
H
TOL
tryptophol
red
sparkling
white
r= 0,62
Verdicchio
Age Age

16. An update on wine ageing and sulfonations Results, indoles
O
OH
N
H S O
O OH
OH
N
H
TOL
tryptophol
red
sparkling
white
O
OH
N
H
OH
indole-3-lactic acid
O
OH
N
H
OH
S
O
O
OH
red
sparkling
white
r = 0,78
Verdicchio

17. An update on wine ageing and sulfonations Results, indoles
O
OH
N
H S O
O OH
OH
N
H
TOL
tryptophol
red
sparkling
white
O
OH
N
H
OH
indole-3-lactic acid
O
OH
N
H
OH
S
O
O
OH
red
sparkling
white
O
OH
N
H
indole-3-acetic acid
O
OH
N
H S O
O OH
red
sparkling
white

18. An update on wine ageing and sulfonations Results, indoles
Amarone SagrantinoTannat
5 6 7 1413109 6 7 1312116 8 10 14 16 19 24 26 31
O
OH
N
H
OH
OH
N
H
TOL
indole-3-lactic acid
tryptophol
OH
N
H
S
O
O
OH
O
OH
N
H
OH
S
O
O
OH
TOL-SO3H
(tryptophol-SO3H)
ILA-SO3H
(indole-lactic acid-SO3H)

19. Results: overviewAn update on wine ageing and sulfonations

20. ConclusionsAn update on wine ageing and sulfonations
Synthesis of TOL-SO3H, ILA-SO3H, IAA-SO3H
Identification of ILA-SO3H, IAA-SO3H
Quantification of TOL-SO3H, ILA-SO3H, IAA-SO3H, ECAT-SO3H, PROC-B2-SO3H
UPLC-MS/MS method development and validation – 32 metabolites
OH
N
H
S
O
O
OH
O
OH
OH
OH
OH
OH
S
O
O
OH
O
OH
N
H S O
O OH
O
OH
N
H
OH
S
O
O
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
S
O
O
OH
TOL-SO3H
tryptophol-SO3H
IAA-SO3H
indole-acetic acid-SO3H
ILA-SO3H
indole-lactic acid-SO3H
ECAT-SO3H
epicatechin-SO3H
ECAT-SO3H
Procyanidin B2-SO3H
Red wines were characterized by ECAT-SO3H, PROC-B2-SO3H
White wines and sparkling was characterized by TOL-SO3H, ILA-SO3H, IAA-SO3H
Aged wines were rich in sulfonates metabolites