Food Bioprocess Technol (2013) 6:719725

DOI 10.1007/s11947-012-0808-7

ORIGINAL PAPER

Effects of UV-C Light Processing on Some Quality

Characteristics of Grape Juices
idem Uysal Pala & Ayegl Krca Toklucu

Received: 26 July 2011 / Accepted: 9 February 2012 / Published online: 24 February 2012
# Springer Science+Business Media, LLC 2012

Abstract White and red grape juices (GJs) were subjected to

ultraviolet C (UV-C) light as a non-thermal preservation technology using a coiled tube UV-C reactor with nine lamps. The
effects of UV-C light on microbial (total aerobic count and
yeast and mould count) and some chemical quality characteristics (total phenolics, antioxidant capacity, anthocyanin and
polymeric colour, etc.) of white and red GJs were investigated.
The results were compared with control (untreated) and heattreated juice samples. Single-pass UV-C treatment (12.6 J/
mL) of white and red GJs resulted in 3.51 and 3.59 log
reductions in total aerobic count and, 2.71 and 2.89 log
reductions in yeast and mould counts, respectively. The microbial loads of both GJs were completely eliminated after
two passes through the reactor (25.2 J/mL). After UV-C and
heat treatments, there were no significant changes in antioxidant capacity, total phenolics, titratable acidity, soluble solids
and pH of white and red GJs (P>0.05). The losses in monomeric anthocyanins were 6.1% and 8.7% after UV-C treatment
of 12.6 and 25.2 J/mL doses, respectively. However, anthocyanin level of red GJ was significantly affected by the heat
treatment with an 11.8% loss (P<0.05). The percent polymeric colour of the red GJ with heat treatment was significantly
higher compared to the colour with the UV-C treatment.
Keywords Ultraviolet C light . Grape juice . Microbial
quality . Total phenolics . Antioxidant capacity .
Anthocyanins
This study was presented as a poster presentation (PS 2.88) at the 2010
EFFoST Annual Meeting Food and Health (1012 November 2010,
Dublin, Ireland).
. U. Pala (*) : A. K. Toklucu
Department of Food Engineering, Faculty of Engineering and
Architecture, anakkale Onsekiz Mart University,
Terzioglu,
anakkale 17020, Turkey
e-mail: cupala@gmail.com

Introduction
Grapes and their juices (GJs) are a rich dietary source of
polyphenols including phenolic acids, flavanols, flavonols,
resveratrol and anthocyanins (Makris et al. 2006; Xia et al.
2010). Due to their high content of polyphenols, they are also
an important source of antioxidants (Dani et al. 2007; Burin et
al. 2010) and have various health-promoting effects based on
their high antioxidant capacity (Park et al. 2009). Thus, there
is a great interest for consumption of both grapes and GJs.
GJ is commonly preserved with thermal pasteurization to
extend its shelf life. However, thermal treatment has adverse
effects on colour and flavour characteristics which are critical factors affecting consumer acceptance (Daoudi et al.
2002). For example, Pozo-Insfran et al. (2006) reported that
24% of anthocyanin pigments were lost during thermal
pasteurization of muscadine grape juice. Thus, new preservation methods are sought by the fruit juice industry. Sulphur dioxide (SO2) can be applied to GJs for controlling the
microbial growth during winemaking (Lorenzini et al.
2010). Since SO2 as a chemical preservative has potential
health risks, the winemaking industry also seeks a new
method instead of SO2 addition to ensure the microbial
stability of GJs and wine (Fredericks et al. 2011).
Emerging non-thermal technologies including high hydrostatic pressure, pulsed electric field, ultrasound (US) and
ultraviolet (UV) light have gained interest to produce microbiologically safe fruit juices and maintain fresh-like characteristics of juices (Patil et al. 2009; Tiwari et al. 2010;
Tikekar et al. 2011). UV processing is a promising nonthermal technology. UV lights, part of the electromagnetic
spectrum in range of 100 to 400 nm wavelengths, are
divided into four regions including UV-A (320400 nm),
UV-B (280320 nm), ultraviolet C (UV-C; 200280 nm)
and vacuum UV (100200 nm) (Koutchma 2009). UV-C
light has germicidal effect on microorganisms such as

720

bacteria, yeasts, moulds and viruses (Tran and Farid 2004;

Keyser et al. 2008; Caminiti et al. 2012). Recent studies
demonstrated that UV-C treatments could be used in preserving liquid food products including various fruit juices
such as orange juice (Tran and Farid 2004; Keyser et al.
2008), apple juice (Caminiti et al. 2012; Keyser et al. 2008;
Franz et al. 2009), grape, cranberry and grapefruit juices
(Guerrero-Beltrn et al. 2009) and pomegranate juice (Uysal
Pala and Krca Toklucu 2011).
Although the antimicrobial effect of UV-C light on GJs
microflora including pathogen and spoilage microorganisms
has been revealed in a few studies (Fredericks et al. 2011;
Lorenzini et al. 2010; Guerrero-Beltrn et al. 2009), no
reports were found on essential quality characteristics of
GJs such as anthocyanins, polymeric colour, phenolics and
antioxidant capacity after UV-C treatment. Thus, objectives
in this study were to determine the effects of UV-C light on
microbial quality of red and white GJs, evaluate the changes
in some chemical quality characteristics (total phenolics,
antioxidant capacity, pH, soluble solids, titratable acidity,
monomeric anthocyanin and polymeric colour) of GJs, and
compare the UV-C and heat treatments in terms of juice
quality.

Materials and Methods

Food Bioprocess Technol (2013) 6:719725

quartz sleeve and nine germicidal UV-C lamps (254 nm,

28 W UV-C output, length 842.4 mm and diameter 16 mm;
Atlantic Ultraviolet Corporation, USA) was used. One of
the UV lamps is located in the quartz sleeve and the other
eight lamps were assembled around the corrugated Teflon
tube (Fig. 1). About 3-L volumes of clarified GJs were
passed through the reactor, and the time for one pass was
150 s (flow rate 20 mL/s). UV intensity per single pass
through the reactor was theoretically calculated using the
flow rate of GJs through the reactor and total output wattage
of UV-C lamps (28 W/lamp 9 lamps) with Eq. 1 suggested
by Geveke (2008) and Keyser et al. (2008):
UV intensityJ=mL Total UV  C output powerW=Flow rateL=s
28  9W=20mL=s;
W J=s
12:6 J=mL per single pass

1
Clarified GJs were passed two times through the reactor,
and sampling was made after each pass. The dosages applied to GJs were, therefore, 12.6 and 25.2 J/mL, respectively. The experiments were performed in duplicate.
For heat treatment, a thermostatic water bath (GFL,
Germany) was used (Turfan et al. 2011; White et al.
2011). Juice samples were filled into 30-mL well-capped
glass tubes and pasteurized at 85 C for 15 min. After
pasteurization, juice samples were immediately cooled to
room temperature.

Juice Preparation
Microbiological Analyses
GJs were prepared from two grape cultivars of avu
(white) and Karasakz/Kuntra (red) harvested from Bozcaada, anakkale, Turkey. After the harvest, grapes were
immediately brought to the laboratory and processed into
juice on the same day. Grapes were washed with tap water
and removed from their stems. About 8 kg of juice were
extracted from 10 kg of grapes by a juicer (Tefal, France)
and filtered using a fourfold cheesecloth. The resulting
juices were depectinized with 2 mL of pectolytic enzyme
(Rapidase C80 MAX) per litre. Depectinization was performed at room temperature for 24 h to increase the initial
microbial loads of the juices for UV-C treatments. Then, the
depectinized white and red GJs were clarified with 3 and
5 mL of 5% bovine gelatin (w/v) per litre at room temperature for 30 min, respectively. The clarified GJs were then
filtered using a filter paper.
UV-C and Heat Treatments
For the UV-C treatment of GJs, a UV reactor (Gentra Stock
Joint Company, Istanbul, Turkey) made of a pump (Jabsco
31295 Series Water Pressure System, USA) as a flow rate
regulator, a stainless steel reflector, a corrugated Teflon tube
(inner diameter 6.35 mm; length 60 m) coiled around

Total aerobic plate (AP) count was determined using serial

dilutions on plate count agar (Merck, Germany) with pour
plate method. Serial dilutions of GJs in the range of 101 to
105 were performed with sterile 0.1% peptone water. The
duplicate plates were incubated at 30 C for 48 h. Enumeration of total yeasts and moulds (YM count) with same
dilutions was carried out on yeast extract, glucose and
chloramphenicol agar (Merck, Germany) at 25 C for 5 days.
Results were expressed as colony-forming units (cfu) per
millilitre (AOAC 2000).
Total Monomeric Anthocyanin Content
Monomeric anthocyanin contents of red GJ samples were
determined using the pH differential method suggested by
Giusti and Wrolstad (2001). For this purpose, red GJ samples were diluted in buffers at pH 1.0 and 4.5, and allowed
to equilibrate for 30 min. The absorbances of each equilibrated
solutions were then measured at 520 nm (max) and 700 nm
for haze correction, using a UVVis spectrophotometer
(Shimadzu 1100, Japan). Calculations were based on malvidin
3-glucoside with molecular weight of 493.5 and extinction
coefficient of 28,000 (Giusti and Wrolstad 2001).

Food Bioprocess Technol (2013) 6:719725

721

Fig. 1 Schematic drawing of

the UV-C reactor including
arrangements of 9 UV-C lamps
(a) and corrugated Teflon tube
(b)

Percent Polymeric Colour

Percent polymeric colour of red GJ was determined according
to the method outlined by Giusti and Wrolstad (2001). Red GJ
samples were diluted with 0.025 M potassium chloride solution to 0.40.6 absorbance value at 520 nm. Then, 0.2 mL of
20% (w/v) potassium metabisulphite solution and 0.2 mL of
distilled H2O were added separately to 2.8 mL of diluted juice
samples. After 15 min, absorbances of the samples were
measured at 420, 520 and 700 nm. Colour density was determined by summing the absorbances of untreated sample with
bisulphite at 420 and 520 nm, while polymeric colour was
determined as the sum of the absorbances of bisulphite-treated
sample at 420 and 520 nm. Turbidity was corrected by measuring absorbance at 700 nm. Percent polymeric colour was
calculated using Eq. 2:

solution was transferred into a cuvette, and increasing aliquots of juice (306090 L) were added to the diluted
ABTS+ solution. Absorbance values were recorded after a
6-min period, and the percentage inhibitions of absorbance
at 734 nm were calculated for each juice volume. Trolox
standard solutions (final concentration 015 mM) were prepared in PBS and assayed at the same conditions. All
measurements were carried out at least three times. The
percentage inhibition values were plotted as a function of
concentration of antioxidants (volume of juice) and Trolox
standards. The TEAC values were calculated from the
slopes of the plots and expressed as micromoles Trolox
per millilitre juice.

5
YMC of White GJ

% Polymeric colour Polymeric colour=Colour density  100

APC of White GJ

Trolox equivalent antioxidant capacity (TEAC) of GJs were

determined based on the ability of a compound to scavenge
the stable ABTS [2,2-azino-bis(3-ethylbenzo-thiazoline-6sulphonic acid)] radical (Re et al. 1999). ABTS radical
cation (ABTS+) was produced by reacting ABTS stock
solution (7 mM) with 2.45 mM potassium persulphate (final
concentration) and allowing the mixture to stand in dark at
room temperature for 1216 h before use. The radical
(ABTS+) solution was diluted with phosphate buffer saline
(PBS, pH 7.4) to an absorbance of 0.700 (0.020) at
734 nm, and the juice was diluted with PBS to give 20
90% inhibition of the blank absorbance. Diluted ABTS+

Log cfu/mL

Trolox Equivalent Antioxidant Capacity

0
Control

UV-C (12.6 J/mL)

UV-C (25.2 J/mL)

Treatment

Fig. 2 Log cfu per millilitre reductions of AP and YM counts in UVC-treated white GJ

722

Food Bioprocess Technol (2013) 6:719725

6
YMC of Red GJ
APC of Red GJ

Log cfu/mL

Ltd., England) and a pH meter (Sartorius PB-11, Germany),

respectively. Titratable acidity was determined according to
the potentiometric method outlined by IFU (1968) and
expressed as grams tartaric acid/100 mL juice.
Statistical Analysis
Results were evaluated using the MIXED procedure of SAS
V8.2 (SAS 1999). The repeated statement and a variance
covariance model (unstructured 1) were utilized to account
for the correlations of all the observations from GJ samples
(Everitt 1995; Jennrich and Schluchter 1986).

Results and Discussion

0
Control

UV-C (12.6 J/mL)

UV-C (25.2 J/mL)

Microbial Quality of UV-C-Treated GJs

Treatment
Fig. 3 Log cfu per millilitre reductions of AP and YM counts in UVC-treated red GJ

Total Phenolic Content

Total phenolic contents of GJs were determined according
to the FolinCiocalteu method suggested by Spanos and
Wrolstad (1990). The juice (100 L) was mixed with
distilled water (900 L), 0.2 N FolinCiocalteu reagents
(5 mL) and 4 mL of saturated sodium carbonate (75 g/L),
respectively. The mixture was vortexed for 1 min and
incubated at room temperature for 2 h in a dark place.
After the incubation process, absorbance of the solution
was measured at 765 nm. Results were expressed as milligrams of gallic acid per litre of juice.
pH, Soluble Solids and Titratable Acidity
Soluble solids (in degrees Brix) and pH were measured at
20 C using an Abbe 5 Refractometer (Bellingham-Stanley

Initial microbial load of clarified white GJ including AP and

YM counts were 4.06 and 3.22 log cfu/mL, respectively. As
seen in Fig. 2, single-pass UV-C treatment (12.6 J/mL UV-C
dose) resulted in a 3.51 log (86.5%) reduction in AP count
and a 2.71 log (84.2%) reduction in YM count, while the AP
and YM counts were totally inactivated after two passes
through the reactor (25.2 J/mL UV-C dose). Initial AP and
YM counts of clarified red GJ were higher (5.59 and 5.24
log cfu/mL, respectively) than white GJ. Single-pass UV-C
treatment of red GJ resulted in a 3.59 log (64.3%) reduction
in AP count and a 2.89 log (55.2%) reduction in YM count
(Fig. 3). AP and YM counts of red GJ were also completely
inactivated after two passes through the UV-C reactor.
The microbial inactivation performance of UV-C treatment
was affected by the red colour of GJ. In a previous study, we
applied UV-C light to clarified pomegranate juice which had
8.3 times higher monomeric anthocyanins than red GJ used in
this study and found only a 0.79 log (23.4%) reduction of AP
count and 0.2 log (10.3%) reduction of YM count after a single
pass through the same reactor with 12.5 J/mL UV-C dose
(Uysal Pala and Krca Toklucu 2011). Guerrero-Beltrn et al.

Table 1 Physico-chemical properties of control, UV-C-treated and heated GJs

Treatment

White GJ

Red GJ
Titratable aciditya

Soluble solids (Bx)

3.9750.07a
3.9800.07a
3.9750.07a

0.2270.01a
0.2270.01a
0.2350.01a

19.150.26a
19.010.26a
19.190.26a

3.390.023a
3.360.023a
3.380.023a

0.3570.021a
0.3630.021a
0.3660.021a

3.9700.07a

0.2310.01a

19.130.26a

3.410.023a

0.4060.021a

Soluble solids

pH

Control
UV-C (12.6 J/mL)
UV-C (25.2 J/mL)

21.880.32ab
21.710.32a
21.630.32a

Heat (85 C, 15 min)

21.810.32a

Column values with different lowercase letters differ (P<0.05)

a

Expressed as grams tartaric acid/100 mL GJ

Results were presented as meansstandard error (n02)

pH

Titratable aciditya

Food Bioprocess Technol (2013) 6:719725

723

Table 2 Total phenols and antioxidant capacity of control, UV-C-treated and heated GJs
Treatment

Control
UV-C (12.6 J/mL)
UV-C (25.2 J/mL)
Heat (85 C, 15 min)

White GJ

Red GJ

Total phenolicsa

Antioxidant capacityb

Total phenolicsa

Antioxidant capacityb

496.2581.04ac
493.7581.04a
488.7581.04a
491.1381.04a

0.6940.1a
0.6740.1a
0.6470.1a
0.6790.1a

820.6035.97a
793.4535.97a
794.0035.97a
809.7535.97a

6.040.41a
5.960.41a
5.900.41a
6.150.41a

Column values with different lowercase letters differ (P<0.05)

a

Expressed as milligrams gallic acid/L GJ

Expressed as micromoles Trolox equivalent per millilitre GJ

Results were presented as meansstandard error (n02)

(2009) reported only a 0.53 log reduction of Saccharomyces

cerevisiae ATCC 10274 in grape juice with deep violet colour
after 30 min of UV-C treatment at 1.02 L/min flow rate using a
UV-C reactor including one lamp with 450 kJ/m2 UV-C dose.
Also, Fredericks et al. (2011) observed a similar inactivation
trend in terms of S. cerevisiae VIN13 for white and red GJs
from cultivars of Chenin Blanc and Shiraz, and indicated that
the efficacy of UV-C light was affected by the colour of juices.
These authors reported 5.38 log and 3.14 log reductions in S.
cerevisiae VIN13-inoculated (about a 6 log level) white and
red GJs after subjecting them to 1.377 J/mL of UV-C dose by a
pilot-scale UV-C system including one lamp, respectively.
Effects of UV-C and Thermal Processing
on Physico-chemical Properties of GJs
pH, soluble solids and titratable acidity values of GJs treated
by UV-C light and heat are presented in Table 1. As seen in
the table, no significant (P>0.05) differences were observed
in these values after UV-C and heat treatments (P>0.05)
compared to the untreated sample. Similarly, Lorenzini et al.
(2010) observed no changes in pH and total acidity of UVC-treated GJ with a SurePure UV reactor (Milnerton, South
Africa). Also, our results are in agreement with other studies
reporting no significant changes in physico-chemical properties of apple juices (Caminiti et al. 2012; Falguera et al.

2011; Noci et al. 2008; Walkling-Ribeiro et al. 2008) and

pomegranate juice (Uysal Pala and Krca Toklucu 2011)
treated with UV-C light.
Effects of UV-C and Thermal Processing on Total Phenolics
and Antioxidant Activity of GJs
Total phenolic levels and TEAC values of GJ samples are
shown in Table 2. As seen, red GJ samples had higher total
phenolics and antioxidant capacity values compared to the
white ones. The higher antioxidant capacity of red GJ may
be attributed to its higher total phenolic content and anthocyanins. A previous study showed that commercial red
grape juices exhibited higher antioxidant capacity than
white grape juices (Davalos et al. 2005).
UV-C and thermal treatments did not affect the total
phenolic levels and TEAC values of juice samples significantly (P>0.05) compared to the untreated juice samples
(Table 2). To our knowledge, there is no report on the effects
of UV-C treatment on total phenolics and antioxidant capacity of GJ. However, there are several reports concerning
the effects of UV-C light on phenol content and antioxidant
capacity of pomegranate and apple juices. Similar to our
results, no significant changes were observed in total phenolic content and TEAC values of pomegranate juice subjected to UV-C light (Uysal Pala and Krca Toklucu 2011).

Table 3 Total monomeric anthocyanin and polymeric colour of control, UV-C treated and heated red GJ
Treatment

Anthocyanina

Colour density

Polymeric colour

Polymeric colour (%)

Control
UV-C (12.6 J/mL)
UV-C (25.2 J/mL)
Heat (85 C, 15 min)

51.371.34ab
48.241.34a
46.881.34a
45.301.34b

1.36130.09a
1.30500.09a
1.30380.09a
1.47380.09a

0.45130.032a
0.43330.032a
0.43250.032a
0.59500.032b

33.110.31a
33.180.31a
33.260.31a
40.370.31b

Column values with different lowercase letters differ (P<0.05)

a

Expressed as milligrams malvidin 3-glucoside per litre GJ

Results were presented as meansstandard error (n02)

724

However, Noci et al. (2008) found a significant decrease in

total phenol content of apple juice after UV-C treatment,
while antioxidant capacity of the juice was not affected
compared to fresh juice. On the other hand, Caminiti et al.
(2012) reported that there were no significant changes in
total phenol content of apple juice exposed to UV-C light,
while TEAC values were decreased as the UV-C dosage was
increased.
Effects of UV-C and Thermal Processing on Anthocyanin
Content and Polymeric Colour of Red GJ
Monomeric anthocyanin content and polymeric colour value
of red GJ treated by UV-C and thermal processes are shown
in Table 3. Monomeric anthocyanin content of red GJ did
not change significantly after UV-C treatment (P>0.05).
The losses in monomeric anthocyanins were only 6.1%
and 8.7% after UV-C treatments of 12.6 and 25.2 J/mL
doses, respectively. Similarly, no significant changes were
found in total anthocyanin content of pomegranate juice
after UV-C treatment (Uysal Pala and Krca Toklucu
2011). The losses in monomeric anthocyanins of pomegranate juice were 3.89% and 8.4% after 37.41 and 62.35-J/mL
doses of UV-C treatments, respectively. However, GuerreroBeltrn et al. (2009) reported that longer residence time of
deep violet-coloured GJ in the UV-C reactor negatively
affected the colour parameters of the juice. For example,
the total colour difference (E) of GJ increased depending
on the treatment time of UV-C light and flow rate of juice in
the reactor. Since photons as radiation energy of UV-C light
are absorbed by organic molecules during UV-C irradiation,
photodegradation of conjugated bonds found in organic
molecules occurs (Koutchma 2009). Therefore, extensive
UV-C light may cause photodegradation of anthocyanins
to colourless pigments.
Thermal treatment significantly affected the anthocyanin
content of red GJ with an 11.8% loss compared to the UV-C
treatment (P<0.05). Thermal processing leads to degradation of anthocyanins accompanying the formation of browncoloured polymeric pigments resulting in colour deterioration (Patras et al. 2010; Cavalcanti et al. 2011). In fact,
polymeric colour of heated GJ was significantly higher than
that of control and UV-C treated samples (P<0.05). Similarly, Uysal Pala and Krca Toklucu (2011) reported that
polymeric colour values of pomegranate juice significantly
increased with thermal treatment, while there were no significant changes in these values after the UV-C treatment.

Conclusions
In this study, GJs from two different grape varieties named
avu (white) and Karasakz/Kuntra (red) were

Food Bioprocess Technol (2013) 6:719725

subjected to UV-C light with 12.6 and 25.2-J/mL doses.

The microbial loads of both GJs were completely inactivated after UV-C treatment of 25.2-J/mL dose (two passes
through the UV reactor). This is the first study reporting the
effects of UV-C treatments on phenolics, antioxidant activity and total anthocyanins of GJ. Phenolics and antioxidant
capacity of GJs were well maintained after the UV-C light
treatment. Moreover, anthocyanin pigments and polymeric
colour which are the main indicators of colour quality of red
GJ were enhanced by UV-C light treatment compared to
thermal treatment. In that respect, we can conclude that UVC light technology is highly promising for prolonging the
shelf life of GJs and could be an alternative to the thermal
treatment.
Acknowledgments The authors thank Mr. Yusuf Koprulu, Gentra
Stock Joint Company (Istanbul, Turkey) for designing and manufacturing the UV system.

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