The Vine is an area within the WBWE, conceived to encourage new businesses that evolve around wine and vine industries. Using every resource, and expanding every possibility, such an initiative shows the great potential that can be developed in the sector. High Pressure Processes Group, Department of Chemical Engineering and Environmental Technology, School of Engineering, University of Valladolid, Spain Instituo de Biologia Experimental Tecnológica, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Portugal corresponding author: amatias@ibet.pt Further information about "The Vine Science Award" on http://www.worldbulkwine.com/thevine/ing/posters.html

1. 8thWorldBulkWineExhibition.Amsterdam,TheNetherlands,21-22November2016
t
(s)
Amplitude
(%)
1 30 10
2 30 55
3 30 100
4 60 10
5 60 55
6 60 100
7 90 10
8 90 55
9 90 100
10 60 55
11 60 55
EXTRACTION KINETICS OF ANTHOCYANINS FROM WINE LEES: IMPACT OF MICROWAVE
AND SONICATION PRETREATMENTS
R. ROMERO-DÍEZ A,B,C, L. RODRIGUES B,C, S. RODRÍGUEZ-ROJO A, C.M.M. DUARTEB,C, MARÍA JOSÉ COCERO A, A. A MATIAS * B,C
A High Pressure Processes Group, Department of Chemical Engineering and Environmental Technology, School of Engineering, University of Valladolid, Spain
B Instituo de Biologia Experimental Tecnológica, C Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Portugal
*corresponding author: amatias@ibet.pt
% H2O
(v/v)
RS-L
(mg/mL)
t
(s)
AC
(mg/gLEES)
Tª
(ºC)
1 10 0.2 90 2.39 106
2 100 0.2 90 0.08 117
3 50 0.15 60 3.12 73
4 10 0.1 30 1.48 53
5 100 0.2 30 0.02 105
6 50 0.15 30 2.42 60
7 100 0.1 90 0.04 68
8 50 0.15 60 2.90 72
9 10 0.2 30 1.49 65
10 50 0.15 60 2.86 73
11 50 0.1 60 2.00 68
12 50 0.2 60 2.18 80
13 10 0.1 90 2.64 77
14 10 0.15 60 1.83 75
15 50 0.15 90 3.74 98
16 100 0.1 30 0.02 38
17 100 0.15 60 0.03 88
Average MP 50 0.15 60 2.96 72
18 37 0.17 90 3.40 115
19 37 0.17 90 3.46 114
Average OP 37 0.17 90 3.46 115
High Pressure Processes Group
WineSense Project has received funding from the People Programme (Marie Curie Actions) of the European Union's Seventh Framework Programme FP7/2007-2013/under REA Grant Agreement nº 612208
Wine industry is an important sector for the EU economy and the development of new wine-related products could lead to sustainable growth and development of the sector.
Winemaking processes generates huge amounts of wastes and by-products, including vine pruning, grape stalks, grape pomace and wine lees. The latter are water-waste residues
generated during maceration and fermentation steps of red and white wine production. These dregs constitute a rich source of bioactive compounds namely (poly)phenols, such as
anthocyanins, which are very useful in industries such as cosmetics, pharmaceutics and food.
This work is part of a major project whose main goal is the development of effective green strategies and processes for the selective recovery of bioactive from winemaking waste
streams. The aim of this study is trying to maximize the extraction of anthocyanins from theses wastes. For this purpose, extraction kinetics have been explored using different
biosolvents . In addition, the influence of two pretreatment (microwaves and sonication) on the final anthocyanin concentration have been studied before solid-liquid extraction.
BACKGROUND
Raw material: - Freeze-dried Porto wine lees
Analytical methods:
- AC (Anthocyanin Content)  extraction curves
- HPLC-DAD-UV (High Performance Liquid Chromatography)
300 rpm
pH = 2.6
180 minutes
Variables of study
Solid-liquid ratio: 0.1, 0.05, 0.03, 0.025 (g/mL)
Solvent: EtOH, three mixtures of EtOH:H2O (v/v)
(75:25, 50:50, 25:75)
Extraction temperature: 25, 35, 45 ºC
Conditions
EXTRACTION KINETICS: SOLID-LIQUID EXTRACTIONS Microwave pretreatment
Three main parameters:
• S-L ratio: 0.20, 0.15, 0.10 (g/mL)
• Solvent mixture: 90:10, 50:50, 0:100 EtOH: H2O
• Time of microwaves pre-treatment: 30, 60, 90s
Sonication pretreatment
A central composite design (CCD) 2^3 with start points
(17 experiments):
• Three levels (-1, 0, 1)
• Triplicate of the central point
Fixed parameters:
• S-L ratio: 0.1 (g/mL)
• Solvent: EtOH:H2O 50:50 (v/v)
Two main variables:
• Amplitude (10, 55,
100%)
• Time of sonication pre-
treatment (30, 60, 90s)
PRETREATMENT + S-L EXTRACTIONS
MATERIALS AND METHODS
Solvents: - Ethanol ( 99.9%) - Distilled water
Microwave pretreatment
AC was measured just after the MW pretreatment. The center
point was repeated in triplicate. The optimum (37% H2O, RS-L:
0.17, t: 90s) was calculating using the corresponding statistical
software and it was also performed in triplicated.
37 % H20 RS-L : 0,17
t : 90 s
Figure 4: principal cause-effect graph for the experimental design for the
MW pretreatment
Table 1: experimental design for MW pretreatment.
RESULTS
0.00
0.20
0.40
0.60
0.80
1.00
1.20
0 25 50 75 100 125 150 175
AC(mgMALDIVIN/gDRYLEES)
Time (min)
Rs-l: 1/10 Rs-l: 1/20 Rs-l: 1/30 Rs-l: 1/40
Figure 1: Study of the influence of the solid-liquid ratio on the final
anthocyanin concentration (Tª:25ºC, 180 min, 100% EtOH)
Solid-liquid extraction curves
Final AC increases when RS-L decreases, but the
use of solvent is increased four times.
0.00
0.50
1.00
1.50
2.00
2.50
3.00
0 25 50 75 100 125 150 175
AC(mgMALDIVIN/gDRYLEES)
Time (min)
100%EtOH 75:25 (v/v) EtOH:H2O
50:50 (v/v) EtOH:H2O 25:75 (v/v) EtOH:H2O
Figure 2: Study of the influence of the solvent mixture on the final
anthocyanin concentration (Tª:25ºC, 180 min, RS-L: 1/10)
Mixtures with less content in water improve
the anthocyanin extraction.
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
0 20 40 60 80 100 120
AC(mgMALDIVIN/gDRYLEES)
Time (min)
Tª: 25ºC Tª: 35ºC Tª: 45ºC
Figure 3: Study of the influence of the temperature on the final
anthocyanin concentration(50:50 (v/v) EtOH:H2O, 180 min, RS-L: 1/10)
Temperature decreases the time of extraction
and increases the final AC of the extracts.
Sonication pretreatment
No significant differences were found between all the experiments. Even
between experiment 1 and 9 which has the minimum and maximum conditions
of the studied variables, respectively, there are not notable differences
(1.49·10-3 and 1.80·10-3, respectively)
Table 2: experimental design
for sonication pretreatment.
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
0 15 30 45 60 75
AC(mgMALDIVIN/gDRYLEES)
Time (min)
Rs-l: 1/10, Tª: 25ºC, 50:50 (v/v) EtOH:H2O
Sonication Pretreatment + S-L Extraction (25ºC)
Optimum MW Pretreatment + S-L Extraction (25ºC
Figure 5: Comparison between a conventional solid-liquid extraction and using microwave and
sonication pretreatments
The use of the pretreatments decrease notably
the time of extraction. MW increase the AC twice.
Solid-liquid extraction curves
Solid-liquid ratio: no significant differences were found between using a ratio of 1/10 or 1/40 (g/mL) that can support the use of four times more solvent. Mixture solvent: using mixtures
with high contents of water do not favor the extraction of anthocyanins. No notable differences were found between using a 50:50 or a 75:25 mixture (v/v) EtOH:H2O, so 50:50 was a better
choice due to the lower consumption of ethanol. Temperature: temperature reduces the time to achieve a determined AC, however 25ºC was selected to perform the S-L extractions after
the pretreatments in order to avoid the degradation of anthocyanins.
Pretreatments influence on the final AC
The use of a microwave pretreatment increases the final AC of the extract approximately two times and decreases the time to obtain a determined AC from 45 minutes to less than 5
minutes. Meanwhile the use of a sonication pretreatment only reduces the time of extraction considerably but the final AC achieved is the same as the one obtain with a conventional S-L
extraction.
CONCLUSIONS