Int.J.Curr.Microbiol.App.

Sci (2018) 7(10): 183-192

International Journal of Current Microbiology and Applied Sciences

ISSN: 2319-7706 Volume 7 Number 10 (2018)
Journal homepage: http://www.ijcmas.com

Original Research Article https://doi.org/10.20546/ijcmas.2018.710.019

Effect of Pretreatments on the Quality of Grape Wine

R. Navarasam*, N. Karpoora Sundara Pandian, B. Dhanalakshmi and M. Abdul Reiyas

Food Technology, Tamil Nadu Veterinary and Animal Sciences University, College of food
and Dairy Technology, Koduveli, Chennai-600052, Tamil Nadu, India

*Corresponding author

ABSTRACT

Keywords This research paper concentrated on studies on the effect of pretreatments on the quality of
grape wine. The changes due to pretreatments on the functional properties are such as total
Red wine, Conventional phenols, tannins and catechins. Hence, various thermal treatments such as microwave with
thermal Pasteurization,
Microwave treatment, a combination of 300 W for 2, 4 and 6 min and conventional thermal pasteurization at 50,
Tannin, Catechin 60, 70°C with the time interval of 30 and 60s were applied to the grape must to study the
extraction efficiency of polyphenols. The quality parameters of prepared wine from the
Article Info
grape must subjected to conventional thermal pasteurization at 70°C for 60 s revealed
Accepted: increase in total phenolic compound and tannin content with mean values of 210.43
04 September 2018 mg/100 ml of wine and 118.27 mg/ 100 ml of wine, respectively whereas microwave
Available Online: treatment for 6 min revealed increase in catechin content with mean value of 68.95 mg/
10 October 2018 100 ml of wine at the end of fermentation.

Introduction and high pressure processing (Liu and Casas,

2010). Since high pressure processing is
Pasteurization is a thermal process in which expensive, conventional thermal
heating of liquid food at particular time and pasteurization and microwave exposure may
successive cooling is performed with the be the choice of pretreatment on grape must in
intention of killing pathogens and deactivating preventing mixed culture fermentation and
enzymes. Grape juice pasteurization even kills extracting the bioactive compounds from the
the most resistant mold which is detrimental grapes towards the production of quality grape
for the production of quality wine (Pederson, wine.
1936).
The applied pre-treatments will inhibit such
Hence, pasteurization of grape must prior to kind of problems and maintain the product
fermentation (the inoculation of yeast) may quality and stability. By introducing the
facilitate the population of yeast in the different levels of dosages of pre-treatments
development of quality wine. like microwave and ultra-violet and blanching,
we can perceive the long shelf-life,
Pasteurization can be accomplished by maintenance of the nutritional content and
conventional heating, microwave exposure quality of grape wine.

183
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

By analyzing the impacts of pre-treatments on Chemicals

the quality of grape wine, we can provide
scientific knowledge on the quality aspects of The following chemicals were used for the
fermented beverages to food industries those quality analysis of grape must and wine
are undergoing fruit processing and includes standard sodium hydroxide,
preservation. Grapes contains high amount of phenolphthalein, methanol, sodium carbonate,
phenolic compounds that possess antioxidant FCR reagent (Folin Ciocalteau Reagent),
property which prevents cancer, it reduces the gallic acid, cuprous sulfate, sodium potassium
risk of strokes and also it boosts-up the tartrate, sodium iodide, sulphuric acid, starch,
immune system. sodium thiosulfate, methylene blue, Fehling A
and B solutions, glucose, lead acetate,
Hence, thermal treatments such as potassium chromate.
conventional thermal pasteurization and
microwave pasteurization on grape must were Glassware
designed to fulfill the targeted objective.
Glass wares viz., burette, pipette, volumetric
Materials and Methods flask, conical flask, measuring cylinder, test
tubes, beaker, petri plates, desiccators,
In this section, materials and methods used in weighing balance, round bottom flask,
the preparations of grape wine, pretreatments digestion flask and sterilized jars were used
and quality evaluation are discussed. for experimentation.

Materials List of devices or instruments used in

experimentation
Grapes
Magnetic stirrer (Make: Remi equipments),
The grapes used in the present study were of pH meter (Make: SUSIMA Technologies
good grade local variety, acidic and procured Private Limited, Chennai), Distillation flask,
from local market. Pycnometer, Hydrometer, Thermometer, Hot
plate, Microwave oven (make: Godrej), UV-
Yeast culture Visible spectrophotometer (Make: UV mini
1240 shimadzo), Distillation unit (Borosil).
To ferment the grape must, commercially
available dry granular brewers‟ yeast procured Methods
from Venus Essence, Chennai. The culture
was stored at 4ºC. Selection of fruits

Sugar Undamaged, healthy ripened grapes having

average pH of 3.0 to 3.6 were selected after
Sucrose was added during the process to manual sorting and discarding of defective
adjust the Total Soluble Solids (TSS). The bunches.
sugar content is expressed as per cent soluble
solids or °Brix. Initial Sugar content of grape Preparation of grape must
must was measured using a refractometer. The
TSS per cent was adjusted to 22°Brix by Undamaged, 20 kg of healthy ripened grape
adding sugar for effective fermentation. berries were de-stemmed and then washed

184
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

with boiled and cooled water. With the help of subjected to thermal exposure at 50, 60 and
fruit juicer (available at the department of 70°C for 30 and 60 s, respectively (Plate 2.2).
Food Plant Operation, CFDT) grape must was
prepared. Grape skins and seeds were kept in Microwave treatment
must itself for extraction of color and phenolic
compounds. The must was kept overnight The prepared grape must was subjected to
under aerobic fermentation. radiative treatments under microwave
exposure (Plate 2.3) at a power level of 300 W
Analysis of grape must with time intervals of 2, 4 and 6 minutes
before addition of starter culture to destroy the
The grape must was analyzed to estimate the harmful microorganisms and extract bioactive
sugar level, pH and amount of phenolic compounds from grape skin and seeds in the
compounds. must.

Preparation of sugar solution Preparation of starter culture

Commercial sugar was procured from the The starter culture was prepared by using 2%
local market. The stock sugar solution was of brewer‟s yeast. It was activated by adding
prepared by dissolving 50g of sugar in 50 ml 2g of yeast into 100 ml of warm water.
of water boiled for 10 minutes and then Activation of yeast was observed by rise in the
allowed to cool at room temperature and used volume of yeast in warm water.
for supplementation in grape must to adjust
the sugar level to 22 °Brix by using hand Alcoholic fermentation of grape must
refractometer.
The pretreated grape must (500 ml) was taken
Pre-fermentation treatments in 1000 ml Erlenmeyer flask and added with
sugar syrup (given in 2.2.4) to adjust the sugar
The extracted grape must was subjected to level at 22°Brix. The prepared must was
pretreatments before its fermentation by the inoculated with 2% starter culture and
yeast to produce ethanol. The grape must was incubated at 28°C. Steps involved in the
subjected to thermal treatments with alcoholic fermentation are presented in figure
conventional pasteurization and microwave 1.
radiation to destroy the pathogens (Table 1).
Post fermentation process
It could help to improve the color intensity
and phenolic contents in the final product. The finished wine contains impurities
including dead cells, protein hazes and
Pasteurization residual sugars and does not have complete
quality (sensory) attributes. Hence, it requires
Pasteurization is the effective heat treatment to post fermentative treatments to make wine
destroy all pathogenic organisms which causes potable. In fact, there are five goals of
spoilage to the product. The temperature is a “finishing” a wine: clarity, stability,
variable that directly affects the growth rate of compositional adjustment, style development
the microorganisms (Charoenchai et al., 1998) and packaging. It is also important to prevent
and the final composition of wine (Torija et unwanted microbial growth in the wine after
al., 2003). Hence, the grape must was primary fermentation (i.e., alcoholic

185
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

fermentation) is complete as this will affect Table 3 shows the tannin content (mg/100 ml)
the flavor and aroma profile. After alcoholic of control and pretreated wine samples. Mean
fermentation, post fermentation processes values of the tannin content (mg/100 ml) in
were carried out. control and pretreated wine samples
(79.88±.09, 61.50±.35, 76.20±.22, 109.98±.19,
Siphoning / Racking 86.18±.26, 94.83±.12, 95.05±.23, 105.70±.12,
100.20±.12 and 118.27±.17) were obtained. It
The flasks or bottles containing prepared was found that there was significant difference
wines were stored in refrigeration and debris (P≤0.01) in the values of the tannin content
called „lees‟ was allowed to settle by gravity. (mg/100ml) of pretreated wine samples with
No settling or fining agent was added. The control sample. The relative change in the
clear supernatant was siphoned and bottled tanning content of treatments with the control
separately. is given in figure 2.

Bottling and Storage Table 4 shows the catechin content (mg/100

ml) of control and pretreated wine samples.
The cleared wine was stored in glass bottles Mean values of the catechin content (mg/100
(sterile bottles) under refrigerated conditions. ml) in control and pretreated wine samples
The sequential operations followed in grape (48.07±.10, 31.03±.20, 44.50±.26, 68.95±.22,
wine production is given in plate 2.4. 30.07±.22, 30.75±.12, 33.33±.20, 36.20±.70,
35.15±.18 and 37.02±.10) were obtained. It
Results and Discussion was found that there were significant (P≤0.01)
changes in values of the catechin content of
Table 2 shows the total phenol content pretreated wine samples with control sample.
(mg/100ml) of control and pretreated samples The relative change in the catechin content of
during wine making. Mean values of total treatments with the control is given in figure
phenol content in control and pretreated 3.
samples (452.28±0.26, 445.58±0.25,
449.27±0.35, 454.88±0.28, 445.23±0.2, Effect of pretreatments on the quality of
446.03±0.26, 450.07±0.23, 450.42±0.27, grape wine
452.60±0.34 and 458.17±0.22) were obtained
during 0th day of wine making. Mean values of Effect on total phenols
total phenol content in control and pretreated
samples (175.12±0.12, 176.62±0.17, The effect of microwave and conventional
178.98±0.12, 190.73±0.22, 176.35±0.19, thermal pasteurization on the quality
178.40±0.17, 182.07±0.22, 190.77±0.3, parameters of grape wine such as total
195.85±0.3 and 210.43±0.3) were obtained phenols, tannin and catechin was studied. It
during 20th day. was observed from table 2 that there was
significant change in the total phenol content
It was found that there was significant of grape must and wine among the treatments
(P≤0.01) difference in the values of total at 1 per cent level. The control recorded the
phenol content of control and pretreated total phenol contents of 452.28mg/100ml of
samples during wine making. There were grape must and 175.12mg/100ml for wine,
significant changes (P≤0.01) in the total respectively. The treatments exposed at 300 W
contents in the treatments during the power levels for 6 min expressed the mean
successive intervals of wine making. total phenol content of 454.88mg/100ml of

186
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

grape must and 190.73 mg/100ml of wine. It mg/100ml, respectively in the total phenolic
was noticed that the microwave exposure for 6 content. The treatments were on par (T6 and
min extracted the phenolic content higher than T7) before fermentation whereas grape wine
the control in the grape must and wine and showed a significant difference at 1 per cent
also interpreted that the increase in microwave level and the increased exposure time
exposure increases the phenolic extraction in improved the total phenolic extraction.
grape must and wine. The results observed
were on par with the results of carew et al., The thermal exposure of grape must at 70°C
(2014). for 30 and 60s recorded 452.60 mg/100 ml
and 458.17 mg/100 ml, respectively whereas
The intervention of conventional thermal grape wine recorded 195.85 mg/100 ml and
pasteurization on the extraction of total phenol 210.43 mg/100 ml, respectively in the total
content from grapes was evaluated at 50, 60 phenolic content and the treatments were on
and 70°C for 30 and 60s exposure time. The par (T8 and T9) before fermentation whereas
thermal exposure of grape must at 50°C for 30 grape wine showed a significant difference at
and 60 s showed a slight change in the total 1 per cent level of significance.
phenolic extraction and the treatments were on
par (T4 and T5) before fermentation whereas Among the treatments, higher total phenolic
grape wine showed a significant difference at extraction was observed on the treatments that
1 per cent level of significance on the phenolic exposed to conventional thermal
content. The increased exposure time pasteurization than the treatments exposed to
improved the total phenolic extraction. The microwave. This may be because of the low
thermal exposure of grape must at 60°C for 30 power level of the microwave considered in
and 60s recorded 450.07 mg/100ml and this study. It was also observed that the total
450.42 mg/100ml, respectively whereas grape phenolic extraction due to the pretreatments
wine recorded 182.07 mg/100ml and 190.77 was superior to control.

Fig.1 Flowchart for alcoholic fermentation of grape must

Pre-treated grape must

↓
Adjust TSS 22°Brix by adding cane sugar
↓
Addition of starter culture
↓
Fermentation at 28°C in BOD incubator under anaerobic condition for about 20 days
↓
Filtration/clarification
↓
Racking and Siphoning
↓
Bottling and Storage

187
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

Fig.2 Comparison of tannin content (mg/100 ml) of wine between control and treatments

Fig.3 Comparison of catechin content (mg/100 ml) of wine between control and treatments

Table.1 Experimental design

Dosages (Independent variables) Quality parameters (Dependent variables)

300 W 2 min(T1)
Microwave 4 min(T2) 1. Total phenols
Treatment 6 min(T3) 2. Tannins
50 °C 30s(T4),60s(T5) 3. Catechins
Pasteurization 60 °C 30s(T6),60s(T7)
70 °C 30s(T8),60s (T9)

188
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

Table.2 Total phenol content of pretreated and control samples during wine fermentation
(Mean±SE)

Treatment 0th day 20th day F-Value

d a
Control 452.28±0.26 175.12±0.12 955710.014**
T1 445.58±0.25a 176.62±0.17d 771444.569**
T2 449.27±0.35b 178.98±0.12c 558303.391**
T3 454.88±0.28e 190.73±0.22f 599834.362**
a b
T4 445.23±0.2 176.35±0.19 749070.011**
a d
T5 446.03±0.26 178.40±0.17 984846.176**
T6 450.07±0.23c 182.07±0.22d 473784.817**
T7 450.42±0.27c 190.77±0.3e 764747.191**
T8 452.60±0.34d 195.85±0.3g 371291.804**
T9 458.17±0.22f 210.43±0.3h 604723.206**
F-Value 241.999** 264.751**
**- Highly significant (P≤0.01) at intervals, a, b, c and ABCDE – superscripts with same letter indicates the treatments are
on par

Table.3 Tannin content (mg/100 ml) of wine (Mean±SE)

Treatment Wine
Control 79.88±.09b
T1 61.50±.35a
T2 76.20±.22e
T3 109.98±.19h
T4 86.18±.26c
T5 94.83±.12d
T6 95.05±.23d
T7 105.70±.12g
T8 100.20±.12f
T9 118.27±.17i
F-Value 6317.294**
a, b, c, d, e, f, g, i
**Highly significant (P≤0.01) at interval, – superscripts with same letter indicates the treatments are on
par.

Table.4 Catechin content (mg/100 ml) of wine (Mean±SE)

Treatment Wine
Control 48.07±.10h
T1 31.03±.20b
T2 44.50±.26g
T3 68.95±.22i
T4 30.07±.22a
T5 30.75±.12ab
T6 33.33±.20c
T7 36.20±.70c
T8 35.15±.18d
T9 37.02±.10f
F-Value 1769.973**
**Highly significant (P≤0.01) at intervals, a, b, c, e, f, g, h, i – superscripts with same letter indicates the treatments are on
par

189
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

Plate.1 Conventional thermal pasteurization treatment

Plate.2 Microwave treatment

Plate.3 Wine making

a) Grapes, b) Blanching, c) grape must, d) Brewer‟s Yeast, e) BOD incubator, f) wine

Effect on tannins was noticed that the microwave exposure for

6 min extracted the tannin content higher than
It was observed from table 4.9 that there was the control in the grape wine. It was also
significant change in the tannin content of interpreted that the increase in microwave
grape wine among the treatments at 1 per cent exposure increases the tannin extraction in
level. The control recorded the tannin grape wine.
contents of 79.88 mg/100ml of grape wine.
The treatments exposed at 300 W power The intervention of conventional thermal
levels for 6 min expressed the mean tannin pasteurization on the extraction of tannin
content of 109.98 mg/100ml of grape wine. It content from grape wine were evaluated at 50,

190
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

60 and 70°C for 30 and 60s exposure time. control in the grape wine. The increased
The thermal exposure of grape must at 50°C exposure time of microwave improved the
for 30 and 60 s showed a slight change in the catechin content in wine.
tannin extraction and the treatments (T6 and
T7) showed a significant difference. The The intervention of conventional thermal
increased exposure time improved the tannin pasteurization on the extraction of catechin
extraction. The thermal exposure of 60°C for content from grape wine were evaluated at 50,
30 and 60 s recorded 95.05 mg/100 ml and 60 and 70°C for 30 and 60s exposure time.
105.70 mg/100 ml of grape wine, respectively
and the treatments (T6 and T7) showed a The thermal exposure of 50°C for 30 and 60s
significant difference. The thermal exposure showed a slight change in the catechin
of 70°C for 30 and 60 s recorded 100.20 extraction and the treatments (T4 and T5) were
mg/100ml and 118.27mg/100ml of grape on par. The increased exposure time
wine, respectively in the tannin extraction. decreased the catechin extraction than the
control. The thermal exposure of 60°C for 30
Among the treatments, higher tannin and 60s recorded 33.33 mg/100 ml and 36.20
extraction was observed on the treatments that mg/100 ml of grape wine, respectively and
exposed to conventional thermal the treatments were on par (T6 and T7). The
pasteurization than the treatments exposed to increased exposure time improved the
microwave. This may be because of the low catechin extraction.
power level of the microwave considered in
this study. It was also found that increased The thermal exposure of 70°C for 30 and 60s
exposure time and dosage level improved the recorded 35.15 mg/100 ml and 37.02 mg/100
tannin extraction. It was also observed that ml of grape wine, respectively in the catechin
the tannin extraction due to the pretreatments extraction and the treatments (T8 and T9)
was superior to control. showed a significant difference. Among the
pretreatments, microwave treated samples
Effect on catechins exposed for 6 min exhibited higher catechin
extraction whereas among the conventional
It was observed from table 4.10 that there was pasteurization treatments 70°C for 60 s
significant change in the catechin content of showed maximum extraction than the others.
grape wine among the treatments at 1 per cent It was also observed that the catechin
level. The control recorded the catechin extraction due to the microwave pretreatments
contents of 48.07mg/100L of grape wine. The was superior to control and conventional
treatments exposed at 300 W power level for thermal pasteurization. The results were
6 min expressed the mean catechin content of confirmed by Clodoveo et al., (2016).
68.95mg/100ml of grape wine whereas
treatments exposed at 300 W power level for While considering the quality parameters of
2 min and 4 min expressed the mean catechin wine such as total phenol content, tannin and
content of grape wine 31.03mg/100ml and catechins, the conventional thermal
44.50mg/100ml of grape wine, respectively. It pasteurization recorded maximum values for
was noticed that the microwave exposure for the total phenols and tannins than the control
6 min extracted the catechin content higher and microwave samples whereas catechin was
than the control in the grape wine. It was found higher in microwave samples than the
found that the microwave exposure for 2 and control and samples exposed to thermal
4 min decreased the catechin content than the pasteurization.

191
 Int.J.Curr.Microbiol.App.Sci (2018) 7(10): 183-192

Acknowledgement growth rates and cell biomass of wine

yeasts. American Journal of Enology
The authors acknowledge the help provided and Viticulture, 49(3): 283-288.
by the Department of Food Plant Operations, Clodoveo, M. L., T. Dipalmo, C. G. Rizzello,
College of Food and Dairy Technology and F. Corbo and P. Crupi, (2016).
Department of Dairy Science, Madras Emerging technology to develop novel
Veterinary College and TRPVB, red winemaking practices: An
Madhavaram, TANUVAS for utilizing the lab overview. Innovative Food Science &
facilities for red wine production and analysis. Emerging Technologies, 38: 41-56.
Liu, J. L., J. F. Yuanand and Z. Q. Zhang,
References 2010. Microwave‐ assisted extraction
optimised with response surface
Carew, A. L., W. Gill, D. C. Close and R. G. methodology and antioxidant activity of
Dambergs, 2014. Microwave polyphenols from hawthorn (Crataegus
maceration with early press off pinnatifida Bge.) fruit. International
improves phenolics and fermentation journal of food science and
kinetics in Pinot Noir. American technology, 45(11): 2400-2406.
Journal of Enology and Viticulture, Pederson, C. S., E. A. Beavens and H. E.
ajev-2014. Goresline, (1936). Preservation of
Casas, L., Mantell, C., Rodriguez, M., Grape Juice. Iv. Pasteurization Of
Martinez, D.L.O., Roldan, E.J., De ory, Juices Or Musts Prepared From Several
I., et al., 2010.Extraction of resveratrol Varieties of Grapes 1‐ 2. Journal of
from the pomace of Palomino fino Food Science, 1(4): 325-335.
grapes by supercritical carbon dioxide. Torija, M. J., N. Rozes, M. Poblet, J. M.
Journal of Food Engineering 96: 304- Guillamón and A. Mas, 2003. Effects of
308. fermentation temperature on the strain
Charoenchai, C., G. H. Fleet and P. A. population of Saccharomyces
Henschke, 1998.Effects of temperature, cerevisiae. International journal of food
pH, and sugar concentration on the microbiology, 80(1): 47-53.

How to cite this article:

Navarasam, R., N. Karpoora Sundara Pandian, B. Dhanalakshmi and Abdul Reiyas, M. 2018.
Effect of Pretreatments on the Quality of Grape Wine. Int.J.Curr.Microbiol.App.Sci. 7(10):
183-192. doi: https://doi.org/10.20546/ijcmas.2018.710.019

192