LWT - Food Science and Technology 79 (2017) 534e542

Contents lists available at ScienceDirect

LWT - Food Science and Technology

journal homepage: www.elsevier.com/locate/lwt

Impact of pulsed electric elds, high hydrostatic pressure, and thermal

pasteurization on selected characteristics of Opuntia dillenii cactus
juice
ger b, Dietrich Knorr c, Salah K. El-Samahy d,
Tamer E. Moussa-Ayoub a, d, Henry Ja
a e, *
Lothar W. Kroh , Sascha Rohn
a t Berlin, Institute of Food Technology and Food Chemistry, Department of Food Chemistry and Analysis, Gustav-Meyer-Allee 25, D-
Technische Universita
13355 Berlin, Germany
b
University of Natural Resources and Life Sciences (BOKU) Vienna, Institute of Food Technology, Muthgasse 18, A-1190 Vienna, Austria
c t Berlin, Institute of Food Technology and Food Chemistry, Department of Food Biotechnology and Food Process Engineering,
Technische Universita
Koenigin-Luise-Str. 22, D-14195 Berlin, Germany
d
Suez Canal University, Agriculture Faculty, Department of Food Technology, 41522 Ismailia, Egypt
e
University of Hamburg, Hamburg School of Food Science, Institute of Food Chemistry, Grindelallee 117, D-20146 Hamburg, Germany

a r t i c l e i n f o a b s t r a c t

Article history: Besides its bioactivity, Opuntia dillenii cactus juice exhibits desirable physicochemical properties. In the
Received 23 August 2016 present study, impact of innovative pasteurization techniques e pulsed electric elds (PEF) and high
Received in revised form hydrostatic pressure (HHP) e on the microbial inactivation, selected physicochemical properties and
29 October 2016
rheological characteristics, ascorbic acid, avonols, betacyanins, and the antioxidant activity was eval-
Accepted 31 October 2016
Available online 2 November 2016
uated in comparison to the non-pasteurized fresh juice. Results showed that PEF and HHP, as well as
thermal pasteurization (TP), reduced the microbial populations found in juice from 103 to less than
10 cfu/mL. The non-Newtonian pseudoplastic ow behaviour of juice was maintained by all three
Keywords:
Opuntia dillenii juice
methods. PEF and HHP resulted in a better retention of ascorbic acid, while TP reduced 22% of the
Pulsed electric elds ascorbic acid content. In contrast, TP caused a slight transformation of isorhamnetin glycosides. PEF and
High hydrostatic pressure HHP maintained antioxidant activity of juice comparatively better than TP.
Pasteurization 2016 Elsevier Ltd. All rights reserved.

1. Introduction Consumers are increasingly demanding minimally processed

Health benets of Opuntia dillenii cactus fruit have been re-
ported in different studies (e.g., Loro, del Rio, & Pe rez-Santana,
1999; Daz Medina, Rodrguez Rodrguez, & Daz Romero, 2007;
Bo hm, 2008). O. dillenii (sometimes misleadingly described as
O. macrorhiza) fruit is rich in betacyanins, to some extent similar to
commercial red beets (Beta vulgaris) (Moussa-Ayoub, El-Samahy,
Rohn, and Kroh (2011). Its juice has desirable characteristics of a
low pH (approx. 3.6), moderate total soluble solids TSS (approx.
10
Brix), plain acidic taste, a plentiful deep red-purple color and a
high content of ascorbic acid. But because of the high viscosity of
the O. dillenii fruit mash, an enzymatic maceration is needed for
O. dillenii fruit juice production (Moussa-Ayoub et al., 2016).
* Corresponding author.
E-mail address: rohn@chemie.uni-hamburg.de (S. Rohn).
healthy foods with more natural avour and color, high quality and
long shelf-life. Therefore, improving quality of the food products
and developing the appropriate technologies used for food pres-
ervation is of key importance. Although an efcient inactivation of
spoilage or pathogenic microorganisms can be achieved, thermal
treatments might have a negative impact. In the meantime, also
emerging and innovative non-thermal preservation technologies
such as pulsed electric elds (PEF), high hydrostatic pressure (HHP),
cold atmospheric plasma, or ultrasound are increasingly considered
for preserving different foods (Barba, Esteve, & Frigola, 2012; Barba
et al., 2015; Knorr et al., 2011). PEF is increasingly used for fruit juice
preservation with products already available on the market. The
short pulses of the applied electric eld impact the permeability of
biological membranes causing reversible or irreversible per-
meabilization depending on the purpose of treatment. For preser-
ving foods, a high electric eld strength in the range of 15e40 kV/
cm is needed to achieve a complete irreversible permeabilization of

http://dx.doi.org/10.1016/j.lwt.2016.10.061
0023-6438/ 2016 Elsevier Ltd. All rights reserved.
 T.E. Moussa-Ayoub et al. / LWT - Food Science and Technology 79 (2017) 534e542
the biological membranes resulting in the inactivation of vegetative
microbial cells (Top, Mathys, Heinz, & Knorr, 2006; Knorr et al.,
2011; Barba et al., 2015). With regard to the use of HHP for the
preservation of high quality foods, a signicant microbial inacti-
vation in food products (e.g., fruit juices) is achieved by a pressure
at 400 MPa or higher even at ambient temperatures within a suf-
cient holding time of several minutes (Castro & Saraiva, 2005;
Patterson, 2005; To p et al., 2006; Knorr et al., 2011; Barba,
Esteve, & Frigola, 2012). Acidic foods (i.e. fruit juices) are particu-
larly good items for high pressure processing and a pressure around
600 MPa is commercially preferred for achieving microbial inacti-
vation (Castro & Saraiva, 2005).
In the present study, the impact of PEF and HHP on improving or
maintaining the fresh characteristics of the fruit juice produced
from O. dillenii cactus was compared to conventional thermal
pasteurization (TP). The microbial load, selected physicochemical
properties, and rheological characteristics, content of selected
bioactive compounds, and the antioxidant activity of the pasteur-
ized juices were determined.
2. Materials and methods
2.1. Plant sample, juice extraction and pasteurization
Samples from Opuntia dillenii cactus fruits were collected from
an experimental eld owned by the Suez Canal University (Ismailia,
Egypt). After washing and removing glochides and distal parts, the
fruits were mashed by interval mixing for 10 s. Then, mash was
macerated using a commercial preparation of pectolytic enzymes
(Fructozym color, Erbsloh Geisenheim AG, Geisenheim, Germany).
A dosage of 200 mL for every kg fruit mash was applied for a holding
time of 60 min at 50
C. Juice was obtained by raising pressure
gradually to a maximum pressure level of 750 MPa using a manual
laboratory juice presser. The collected juice was diluted with
deionized water (juice: water ratio 2:3) and divided into four parts.
Three parts were treated directly with pulsed electric elds (PEF),
high hydrostatic pressure (HHP), or thermal pasteurization (TP).
The fourth part was used as a control sample.
I) PEF was conducted using a laboratory continuous system with
juice ow rate of 6.6 L/h and an electric eld strength of 35 kV/
cm. The system consisted of a 7 kW pulse modulator (Scandi-
nova Systems AB, Uppsala, Sweden) providing rectangular
monopolar pulses. A pulse width of 3 ms was used (frequency
45 Hz, pulse energy 3.5 J, current 58 A, voltage 21 kV). The juice
was pre-heated from room temperature to 45
C. An outlet
temperature of 67
C after PEF treatment was measured as a
result of the dissipation of electrical energy based on a total
specic energy input of 85 kJ/kg applied during the treatment. A
co-linear treatment chamber was used. A detailed description
including information on electric eld strength distribution and
ow characteristics can be found in Ja ger, Meneses, and Knorr
(2009).
II) HHP was conducted with a high pressure processing batch
system U4000 (Institute for High Pressure Physics, Warsaw,
Poland; Year of manufacture 2003; Max pressure of 800 MPa;
Vessel volume of 0.75 L; Temperature range 25e100
C). The
juice was lled in sterilized plastic bottles and then putted in
vacuumed plastic bags before treatment. A 1:1 mixture of
deionized water and propylene-glycol was used as pressure-
transmitting medium. The juice was subjected to pressure of
600 MPa at ambient temperature for a holding time of 10 min.
The pressure vessel was at ambient temperature, juice tem-
perature was 15
C before pressure was built-up. Due to the
adiabatic heat of compression, the temperature of the juice
535
reached 35
C under pressure and decreased slowly to 28
C
before pressure release after the holding time.
III) TP was conducted using continuous heating lab-scale system
with a juice ow rate of 6.6 L/h. The pre-heating temperature of
juice was 55
C, followed by pasteurization at 95
C of with a
holding time of 3 min. Samples were collected in sterilized glass
bottles.
2.2. Chemicals and reagents
The avonol standards were purchased from Extrasynthese
(Genay, France). HPLC solvents and further chemicals were pur-
chased from Carl Roth GmbH & Co. KG (Karlsruhe, Germany).
2.3. Physicochemical properties
The pH and total soluble solids (TSS) was determined directly in
the non-pasteurized and treated juices. Color parameters were
measured directly in all juices using a Minolta Chroma Meter CR-
300 (Tokyo, Japan). The results were expressed as tristimulus pa-
rameters (L*, a* and b*), where L* (indicates lightness), a* (red-
green) and b* (yellow-blue).
2.4. Ascorbic acid content
As described by Moussa-Ayoub et al. (2016), an HPLC system,
equipped with a simple pump, a UV detector (Knauer Wissen-
schaftliche Gerate GmbH, Germany) and a 250  4 mm Hypersil
ODS column (Knauer, Germany), applying a tetrabutylammonium
hydrogen sulfate/ddH2O/methanol mixture as eluent at a ow of
1 mL/min, was used. Each juice sample was diluted (1:3) with meta-
phosphoric acid (6%), vortexed and ltered prior to injection. An
aliquot of dilution (20 mL) was applied. The ascorbic acid content
was determined at a wavelength of 254 nm and expressed as mg/
100 mL of fresh juice.
2.5. Rheological measurements
Rheological measurements were carried out using Rheometer
MCR301 (Physica, Anton Paar GmbH, Graz, Austria-Europe)
equipped with a CC27/P1 cylindrical measuring system. Analysis
of date was carried out using the software Rheoplus/32 Multi6
V3.40. The measurements were carried out at 20
C. Shear rate test
was performed at shear rate range of 0.1e100/s. The measurement
time of each sample was 60 s with 30 measurement points (2 s for
each measurement point). The ow behaviour data was produced
according to the OSTWALD-DE WAELE model: t K$gn, and the
apparent viscosity (heff) was calculated at shear rate 100/s by the
equation: heff t/g K$gn1, where: t shear stress (mPa),
g shear rate (1/s), K consistency index (mPa.s) and n ow
index.
2.6. Microbiologicial analyses
Yeasts and molds, acid tolerant microorganisms, and total col-
ony count were determined in the non-pasteurized diluted juice
and in the pasteurized juices after a storage at 8
C for 15 d. A series
of dilution up to 103 was made and 500 mL of diluted sample were
plated on plate count agar for total colony count, orange serum agar
for acid tolerant microorganisms, and oxytetracyline glucose yeast
extract agar for determining yeasts and molds. Samples were
incubated for 48 h at 25
C (yeasts) and 32
C for total colony count
and acid tolerant microorganisms. The number of colony forming
units was counted manually and the inactivation effect was
536 T.E. Moussa-Ayoub et al. / LWT - Food Science and Technology 79 (2017) 534e542
expressed as log-cycles of microbial reduction in cfu/mL.
2.7. Extraction for analysis
Analysis of avonols, betacyanins, and antioxidant activity were
carried out with extracts of the freeze-dried material. Fifty mg of
sample per 1 mL aqueous methanol (70%) were sonicated for
15 min and vortexed. Prior to the analyses, all extracts were vor-
texed again and centrifuged at 10,000xg for 10 min (Hermle
Z233MK, Hermle Labortechnik GmbH, Wehingen, Germany). The
supernatants were used for the analyses.
2.8. Identication and quantication of avonols
All supernatants were sonicated for further 5 min, ltrated
through syringe lters (nylon, 0.45 mm, Carl Roth GmbH & Co. KG,
Karlsruhe, Germany) prior to HPLC-DAD analysis. The avonol
glycosides were quantied using HPLC-DAD analysis according to
the method and conditions described by Moussa-Ayoub et al.
(2011). The detection was performed at the three wavelengths
365, 325 and 280 nm, and the avonol content was expressed as
mg/g (dry weight) of the lyophilized juice.
2.9. Determination of the total betacyanins
Betacyanin content in supernatants was calculated on the basis
of a photometric quantication as described by Moussa-Ayoub et al.
(2011). The maximum absorbance for violet to red betacyanins is at
535 nm. Total betacyanins measured at 535 nm was expressed in
mg/g (dry weight) of the lyophilized juice.
2.10. Determination of the antioxidant activity
The degradation of the stable synthetic radical Fremy's salt
(potassium nitrosodisulfonate) in presence of antioxidants in the
sample extracts (50 mg/mL aqueous methanol) was monitored
using electron paramagnetic resonance (EPR) spectrometry as
applied by Moussa-Ayoub et al. (2014). Appropriate extract di-
lutions (1:15) were prepared and 100 mL aliquots were allowed to
react for 5 min with an equal volume of a solution of Fremy's salt
(1 mM in phosphate buffer, pH 7.4). EPR spectra of Fremy's radical
were obtained with a Miniscope MS100 spectrometer (Magnettech
GmbH, Berlin, Germany). The antioxidant activity expressed as mM
Fremy's salt reduced by 100 mL diluted extract, was calculated by
comparison with a control reaction with 100 mL Fremy's salt 1 mM
and 100 mL phosphate buffer.
2.11. Statistical analysis
All analytical data included four replicates and were statistically
evaluated by a one-factorial ANOVA using Tukey's HSD test. Means
having the same letter are not signicantly different at p  0.05.
Table 1
Technofunctional characteristics of the non-pasteurized and the PEF, HHP, and thermally pasteurized juices.
Non- pasteurized
pH 3.68 0.02
Conductivity 3.78 0.02
TSS 4.2 0.1
Color
A 7.47 0.20
B 2.61 0.09
L 21.72 0.18
3. Results and discussion
3.1. Physicochemical properties
The diluted juice recovered from O. dillenii fruits exhibited a low
pH value of approx. 3.6e3.7, placing it in the category of high acidic
foods (<4.5). Further, TSS value was 4.2
Brix (non-diluted: 10
Brix)
(Table 1). Also, Daz Medina et al. (2007) reported that O. dillenii
fruits exhibited a pH and TSS values of approx. 3.3 and 10.3
Brix, in
a contrast to mean pH and TSS values of approx. 6.3 and 14.6
Brix in
fruits from O. cus-indica. Results showed that PEF and HHP caused
a slight decrease in the pH value compared to the non-pasteurized
(control) juice. The produced juices had TSS values almost similar to
that of control juice (Table 1).
With regard to color attributes (L*, a* and b*), the plentiful
purple-red color of O. dillenii fruit juice is due to its high content of
betacyanins. In particular betanin, being the predominant beta-
cyanin in red beets (Beta vulgaris) and fruits from O. dillenii and
O. cus-indica (Moussa-Ayoub et al., 2011). In the present study,
despite of enzymatic hydrolysis and the dilution of the juice, the
nal O. dilenii juice still had a high color impact. With regard to
impact of preservation methodology applied in the present study,
b* values of all juices were almost similar, but slight increases in a*
and L* values of all juices produced in comparison to the non-
pasteurized (control) juice were found (Table 1).
3.2. Ascorbic acid content
In the present study, the non-pasteurized diluted juice had a
mean content of ascorbic acid (23.8 mg/100 mL) (Fig. 1). By
considering the impact of dilution, the juice recovered from the
O. dillenii fruits had a mean content of ascorbic acid (59.5 mg/
100 mL). Moussa-Ayoub et al. (2016) reported that juice recovered
from the O. dillenii fruits had a mean content of ascorbic acid
(55.8 mg/100 mL) being approx. 2.4-fold higher than mean con-
tents in the O. cus-indica cactus pulps as reported by El-Samahy,
Abd El-Hady, Habiba, and Moussa-Ayoub (2006). Daz Medina
et al. (2007) described that O. dillenii fruit had an ascorbic acid
content 1.7-fold higher than O. cus-indica fruits. However, this
O. dillenii juice had higher ascorbic acid content compared to that
found in O. stricta cactus fruits (35 mg/100 g), as published by
Castellar, Solano, and Obon, (2012). In comparison to common fruit
juices such as orange juice (approx. 55 mg/100 mL; Min, Jin, Min,
Yeom, & Zhang, 2003; Elez-Martnez & Martn-Belloso, 2007),
strawberry juice of approx. 65 mg/100 mL (Odriozola-Serrano,
Soliva-Fortuny, & Martn-Belloso, 2008), or strawberry puree of
approx. 63 mg/10 g dry weight (Patras, Brunton, Da Pieve, & Butler,
2009), the O. dillenii juice might be a potent natural source of
vitamin C. With regard to food processing, vitamin C is an impor-
tant food additive in the commercial production of juices and
various food products, where its property to act as reducing agent
offers the protection for some sensitive food ingredients. For
instance, betacyanins are more stable in presence of signicant
PEF HHP Thermally pasteurized
3.65 0.02 3.65 0.01 3.68 0.01
3.70 0.02 3.78 0.01 3.77 0.01
4.1 0.1 4.2 0.1 4.2 0.1
7.65 025 7.92 0.28 7.88 0.54
2.61 0.07 2.62 0.13 2.60 0.28
21.86 0.20 21.93 0.24 21.92 0.76
 T.E. Moussa-Ayoub et al. / LWT - Food Science and Technology 79 (2017) 534e542 537

Fig. 1. Ascorbic acid content of cactus O. dillenii juices depending on pasteurization treatment [mg/mL]. Treated with pulsed electric elds (PEF) or high hydrostatic pressure (HHP).

ascorbic acid concentrations (Herbach, Stintzing, & Carle, 2006). followed by diluting the juice with water caused a sharp decrease in
However, vitamin C is not a stable reducing agent during thermal apparent viscosity (Table 2, Fig. 2A). Despite the impacts of enzy-
treatments in the presence of oxygen (Smuda & Glomb, 2013). matic hydrolysis and dilution on the juice viscosity, the shear stress
While PEF and HHP preserved approx. 98% and 96% of the ascorbic (t) vs. shear rate (g) curves showed that non-treated, hydrolyzed,
acid, respectively, TP resulted in a degradation of approx. 22% of the and even diluted O. dillenii fruit juice still exhibited a non-
ascorbic acid content (Fig. 1). Min et al. (2003) reported that PEF Newtonian pseudoplastic behaviour with an apparent viscosity of
processing did not signicantly change the concentration of 4 mPa s at a shear rate of 100/s (Fig. 2).
ascorbic acid in orange juice, while thermal processing reduced 19% The preservation methods applied in the present study caused a
of the vitamin. slight decrease in the juice viscosity compared to the non-
A number of studies reported that vitamin C is even better pasteurized diluted juice (Table 2, Fig. 2). Similar to juice viscos-
preserved by HHP. Sa nchez-Moreno et al. (2005) reported that PEF ity, results in Table 2 showed that consistency of O. dillenii juice was
(35 Kv/cm) and HHP (400 MPa) processing technologies posed a reduced by maceration and also further by dilution. Obviously, PEF
better retention of ascorbic acid in orange juice compared to treatment did not change the consistency of juice (Table 2). Results
thermal pasteurization (90
C). Results obtained by Barba, Esteve, in Fig. 2B showed also that both juices treated with HHP and TP
and Frigola (2010) showed that thermal pasteurization (90e98
C exhibited a higher viscosity measured at a very low shear rate (0.1/
for 15e21 s) and HHP treatments (100e400 MPa for 2e9 min) s). Their viscosities were 77.1 and 72.4 mPa s, respectively. This
caused a maximum degradation of approx. 12 and 9% of the might explain the decrease in ow index compared to PEF (Table 2).
ascorbic acid in a blended vegetable beverage, respectively. This
research group published several reviews in which further aspects 3.4. Microbial inactivation
on ascorbic acid retention following treatment with HHP and PEF
are described comprehensively (e.g., Barba et al., 2012, 2015). In the present study, the diluted O. dillenii juice had an initial
microbial load higher than 3-log cycle of colony-forming units (cfu
3.3. Rheological properties per mL for total plate counts, yeasts/molds, and acid-tolerant mi-
croorganisms (Table 3). O. dillenii juice can be categorized within
O. cus-indica fruit's pulp or its processed juices exhibit a ow of the acidic foods with pH values lower than (4.5). Therefore, TP at
non-Newtonian pseudoplastic behaviour with a ow index >1
tting well within the OSTWALD-DE WAELE model (El-Samahy, AbdEl-
Table 2
Hady, Habiba, & Moussa, 2006, 2007). As expected, the steady shear
Selected rheological characteristics (according to OSTWALD-DE WAELE model) of the
measurements performed in the present study illustrated that the O. dillenii fruit juices.
juice from O. dillenii behaved also as a non-Newtonian pseudo-
K n R heff
plastic uid with a ow index of 0.66 (Table 2).
O. dillenii fruit juice exhibits a high viscosity (Moussa-Ayoub Before enzymatic hydrolysis 246.51 0.661 0.996 49.87
After enzymatic hydrolysis 68.00 0.620 0.999 11.84
et al., 2016). This high viscosity of O. dillenii juice might result
Non-pasteurized (diluted) 13.70 0.712 0.992 3.97
from its high total ber content consisting of pectin, mucilage, and PEF 14.00 0.688 0.984 3.78
further hydrocolloids. Daz Medina et al. (2007) reported a higher HHP 18.60 0.617 0.972 3.77
total ber content accounted for up to 9.5% of O. dillenii fruit fresh Thermally pasteurized 18.30 0.629 0.973 3.87
weight compared with O. cus-indica. K consistency [mPa.sn]; n ow index; R correlation coefcient; heff effective
Results showed that maceration using pectolytic enzymes viscosity at shear rate 100/s [mPa.s].
538 T.E. Moussa-Ayoub et al. / LWT - Food Science and Technology 79 (2017) 534e542

Fig. 2. A and B. Viscosity versus share rate ow curves of cactus O. dillenii juices depending on pasteurization treatment. (A) Before (lled diamonds) and after enzymatic hydrolysis
(non-lled triangles) and after dilution with water (lled triangles); (B) before (lled diamonds) and after pasteurization of the diluted juice (treated with pulsed electric elds
(PEF), non-lled triangles; treated with high hydrostatic pressure (HHP), lled triangles; thermally treated, lled circles).

Table 3
Microbial load in the O. dillenii juices after storage at 8
C for 15 days [cfu/mL].

Total colony count Yeasts/molds Acid tolerant microorganisms

Non-pasteurized 6,49E 03 4,23E 03 4,27E 03

PEF n.d. n.d. n.d.
HHP n.d. n.d. n.d.
Thermally pasteurized n.d. n.d. n.d.

n.d. not detected (<10 cfu/mL).

 T.E. Moussa-Ayoub et al. / LWT - Food Science and Technology 79 (2017) 534e542
95
C for only 3 min was applied in the present study. The acidity
already offers a kind of protection against the germination of mi-
crobial spores. On the other hand, acidic foods (i.e. fruit juices) are
particularly good items for high pressure processing, and a pressure
of 600 MPa is often preferred for achieving microbial inactivation in
food products (Castro & Saraiva, 2005). To detect the efcacy of the
applied processing methods for achieving a shelf-stable product,
microbial analyses were carried out following a storage period (8
C
of 15 d). PEF, HHP, and TP effectively reduced the initial microbial
populations to below 1-log cycle (Table 3). These results are in
agreement with previous studies. For instance, Yeom, Streaker,
Zhang, and Min (2000) reported that high electric eld strengths
(35 kv/cm) were efcient to reduce about 7-log cycles of either total
aerobic plate count or yeasts/molds found in orange juice to less
than 1 log cfu/mL. Also, Min et al. (2003) reported that either PEF
(40 kV/cm) or TP (90
C for 90 s) reduced the total aerobic plate
count and yeasts and molds to less than 10 cfu/mL. With regard to
HHP processing, the antimicrobial effect can be achieved by
applying HHP at pressures lower than 600 MPa, depending on
different factors (i.e. food matrix). For instance, Zou et al. (2016)
showed that treating mulberry juice with HHP (at 500 MPa) for
only 5 min caused a complete inactivation of aerobic bacteria and
yeasts/molds.
3.5. Flavonol content
While cactus O. cus-indica fruit's peel has different iso-
rhamnetin glycosides with isorhamnetin-3-O-rutinoside as pre-
dominant avonol, O. dillenii fruit's peel has only isorhamnetin-3-
O-rutinoside (Moussa-Ayoub et al., 2011, 2014). Therefore, iso-
rhamnetin glycosides might serve as probable ngerprint of
Opuntia cactus products. In the present study, HPLC-DAD mea-
surements showed that O. dillenii juice contained isorhamnetin-3-
O-rutinoside in an average concentration of 0.45 mg/g of dry
weight (Fig. 3A). Flavonols may have a benecial impact on
lowering the risk of developing degenerative diseases (e.g., Huxley
& Neil, 2003; No thlings et al., 2008). Positive pharmacological
impacts of isorhamnetin or its glycosides isorhamnetin-3-O-ruti-
noside is reported (e.g., Bao & Lou, 2006; Boubaker et al., 2011;
Zhang et al., 2011).
All processing technologies applied in the present study caused
a signicant increase in the content of isorhamnetin-3-O-rutino-
side in comparison to the non-pasteurized juice (Fig. 3A). This can
be also further enhanced by an improved cell disintegration prior to
juice pressing (Moussa et al., 2016). For instance, Odriozola-Serrano
et al. (2008) reported a tendentiously increase in avonols content
of strawberry juice. Moreover, their results suggest that avonols
seem to be stable towards thermal treatments with no signicant
difference in avonol content being observed between fresh, PEF-
treated (35 kv/cm), or thermally (90
C for 30 or 60 s) processed
strawberry juices. With regard to HHP processing, S anchez-Moreno
et al. (2005) reported that HHP signicantly increased the total
avanone content of orange juice. However, no signicant differ-
ence between PEF, HHP, and thermally pasteurized-juices in
avonol content was observed in the present study.
3.6. Betacyanins content
O. dillenii fruit has an intensive purple-red color impact resulting
from their high content of betacyanins with betanin as the pre-
dominant compound. Further, methanolic extracts had mean total
contents of betacyanins more/or less similar to that found in red
beets (Beta vulgaris) being approx. 7-fold higher than contents
found in red fruits from O. cus-indica (Moussa-Ayoub et al., 2011).
In the present study, the methanolic extract of the lyophilized
539
samples from the diluted, non-pasteurized O. dillenii juice had a
mean total betacyanins content of 4.1 mg/g dry weight (Fig. 3B).
With regard to the impact of the PEF, HHP, and TP on the total
betacyanins content in juice, results in Fig. 3B illustrate that the
content of betacyanins remained comparatively stable following
the treatments. This correlates quite well with the observations
made during the evaluation of the color (Table 1). However, infor-
mation about impact of innovative emerging non-thermal tech-
nologies on Opuntia cactus fruits is very rare. Moussa-Ayoub et al.
(2016) reported that mild PEF pre-treatment (3 Kv/cm) protected
the betacyanins and their pattern in the O. dillenii juice compared to
thermal pre-treatment. Improving extraction of betacyanins from
red beets (Beta vulgaris) by a mild PEF treatment was reported by
Loginova, Lebovka, and Vorobiev (2011). The same group showed
also an enhancement of the extraction of red colorants from red
prickly pear (Opuntia stricta Haw.) peels and pulps (Koubaa et al.,
2016).
Stability of betacyanins is affected by different factors such as
their concentration, degree of glycosylation, degree of acylation,
plant matrix, moisture, pH value, temperature, available antioxi-
dants, etc (Herbach et al., 2006). As already mentioned above,
O. dillenii juice has a low pH value and high levels of ascorbic acid,
and hydrocolloids which might offer a kind of protection for
betacyanins and might impact positively their stability during
processing.
3.7. Antioxidant activity
Electron paramagnetic resonance (EPR) spectroscopy illustrated
a high antioxidant activity of O. dillenii fruit or its juice in com-
parison to O. cus-indica (Moussa-Ayoub et al., 2011, 2014, 2016). In
the present study, while PEF and HHP resulted in an antioxidant
activity quite similar and close to the antioxidant activity of the
non-pasteurized juice, TP tends to affect the antioxidant activity
(Fig. 4). With regard to impact of processing technology on the
antioxidant activity of certain products, information in the litera-
ture widely differs. Odriozola-Serrano et al. (2008) reported that
PEF processing technology impacted positively the health-related
bioactive compounds in comparison to conventional TP of straw-
berry juice, but this was not reected in a comparatively higher
antioxidant activity. Sa nchez-Moreno et al. (2005) reported that
HHP and PEF were more effective than TP in preserving bioactive
compounds and the antioxidant capacity of the freshly squeezed
orange juice. The same was shown by Zulueta, Barba, Esteve, &
Frigola (2013) for an orange juice-milk beverage treated by equiv-
alent thermal and non-thermal processes for mild pasteurization.
However, they found an increase in antioxidant activity when
storage time was increased, independently of the preservation
treatment applied.
In this context it might be worth to mention that in the presence
of nitrogen compounds (i.e. amino acids) and reducing sugars or
ascorbic acid, MAILLARD reactions might occur. As mentioned above,
while ascorbic acid might offer a protection for other sensitive
bioactive compounds such as betacyanins either during exposure to
thermal processing or during the product's shelf life (Herbach et al.,
2006), it might be also degraded during the MAILLARD reaction pro-
ducing non-enzymatic browning products (Smuda & Glomb, 2013).
Depending on the amount of MAILLARD reaction products formed,
these might also contribute to a higher antioxidant activity
(Kanzler, Haase, & Kroh, 2014; Yilmaz & Toledo, 2005). Preliminary
results showed that a thermal pasteurization of a cactus fruit juice
at 95
C (for 25 min) resulted in occurring of 3-deoyhexosulose
(data not shown). End products of the MAILLARD reaction e mela-
noidins e have also antioxidant and antimicrobial activities.
Rua n-Henares and Morales (2007) reported that they could
540 T.E. Moussa-Ayoub et al. / LWT - Food Science and Technology 79 (2017) 534e542

Fig. 3. A and B. (A) Total avonol content of cactus O. dillenii juices depending on pasteurization treatment [mg/g dry weight]; (B) Total betacyanins content as measured at 535 nm
of cactus O. dillenii juices depending on pasteurization treatment [mg/g dry weight]. Treated with pulsed electric elds (PEF) or high hydrostatic pressure (HHP).

increase food shelf-life by decreasing oxidation and microbial
growth. Therefore, studies concerning non-enzymatic browning in
Opuntia cactus fruit products are still not sufcient.
4. Conclusion
Results obtained by the present study emphasized the
bioactivity of O. dillenii cactus fruit and consequently, its products.
Especially, the physicochemical and rheological properties place
O. dillenii juice as a promising cactus product. The juice might
improve the sensory attributes of low viscous, pale products such as
processed strawberries. The emerging technologies PEF and HHP, as
applied in the present study produced juice with the same inacti-
vation effect on the microorganisms found in the thermally
 T.E. Moussa-Ayoub et al. / LWT - Food Science and Technology 79 (2017) 534e542
Fig. 4. Antioxidant activity of cactus O. dillenii juices depending on pasteurization treatment. Degradation of the free radical Fremy's salt [%] after reacting with cactus juices for
16 min. Treated with pulsed electric elds (PEF) or high hydrostatic pressure (HHP).
pasteurized juice taking into account a storage period of 15 d. Be-
sides their gentle impact on physicochemical and rheological
properties, PEF and HHP processing further showed a higher degree
of retention for sensitive bioactive compounds and the antioxidant
activity of the fresh O. dillenii juice.
Acknowledgment
This work was partially nanced and supported by the Egyptian
Ministry of Higher Education & Scientic Research. Grateful ac-
knowledgments and thanks go out to Prof. Dr. Bernhard Senge and
t Berlin, Department
Dr. Reinhard Blochwitz, Technische Universita
of Food Rheology for helping with the rheological characterization.
References
Bao, M., & Lou, Y. (2006). Isorhamnetin prevent endothelial cell injuries from
oxidized LDL via activation of p38 MAPK. European Journal of Pharmacology, 547,
22e30.
Barba, F. J., Esteve, M. J., & Frigola, A. (2010). Ascorbic acid is the only bioactive that
is better preserved by high hydrostatic pressure than by thermal treatment of a
vegetable beverage. Journal of Agricultural and Food Chemistry, 58,
10070e10075.
Barba, F. J., Esteve, M. J., & Frigola, A. (2012). High pressure treatment effect on
physicochemical and nutritional properties of uid foods during storage: A
review. Comprehensive Reviews in Food Science and Food Safety, 11, 307e322.
Barba, F. J., Parniakov, O., Pereira, S. A., Wiktor, A., Grimi, N., Boussetta, N., et al.
(2015). Current applications and new opportunities for the use of pulsed
electric elds in food science and industry. Food Research International, 77,
773e798.
hm, H. (2008). Opuntia dillenii - an interesting and promising Cactaceae Taxon.
Bo
Journal of the Professional Association for Cactus Development, 10, 148e170.
Boubaker, J., Bhouri, W., Ben Sghaier, M., Ghedira, K., Dijoux Franka, M. G., & Ghekir-
Ghedira, L. (2011). Ethyl acetate extract and its major constituent, isorhamnetin
3-O-rutinoside, from Nitraria retusa leaves, promote apoptosis of human mye-
logenous erythroleukaemia cells. Cell Proliferation, 44, 453e461.
Castellar, M. R., Solano, F., & Obo n, J. M. (2012). Betacyanin and other antioxidants
production during growth of Opuntia stricta (Haw.) fruits. Plant Foods for Human
Nutrition, 67, 337e343.
Castro, S. M., & Saraiva, J. A. (2005). High-pressure processing of fruits and fruit
products. In S. Da-Wen (Ed.), Emerging technologies for food processing (pp.
65e76). Academic Press.
541
Daz Medina, E. M., Rodrguez Rodrguez, E. M., & Daz Romero, C. (2007). Chemical
characterization of Opuntia dillenii and Opuntia cus-indica fruits. Food Chem-
istry, 103, 38e45.
El-Samahy, S. K., Abd El-Hady, E. A., Habiba, R. A., & Moussa, T. E. (2006). Chemical
and rheological characteristics of orange-yellow cactus pear pulp from Egypt.
Journal of the Professional Association for Cactus Development, 8, 39e51.
El-Samahy, S. K., Abd El-Hady, E. A., Habiba, R. A., & Moussa-Ayoub, T. E. (2007).
Cactus pear sheet and pasteurized and sterilized cactus pear juices. Journal of
the Professional Association for Cactus Development, 9, 148e164.
Elez-Martnez, P., & Martn-Belloso, O. (2007). Effect of high intensity pulsed electric
eld processing conditions on vitamin C and antioxidant capacity of orange
juice and gazpacho, a cold vegetable soup. Food Chemistry, 102, 201e209.
Herbach, K. M., Stintzing, F. C., & Carle, R. (2006). Betalain stability and degradation
e structural and chromatic aspects. Journal of Food Science, 71, 41e50.
Huxley, R. R., & Neil, H. A. W. (2003). The relation between dietary avonol intake
and coronary heart disease mortality: A meta-analysis of prospective cohort
studies. European Journal of Clinical Nutrition, 57, 904e908.
ger, H., Meneses, N., & Knorr, D. (2009). Impact of PEF treatment inhomogeneity
Ja
such as electric eld distribution, ow characteristics and temperature effects
on the inactivation of E. coli and milk alkaline phosphatase. Innovative Food
Science & Emerging Technologies, 10, 470e480.
Kanzler, C., Haase, P. T., & Kroh, L. W. (2014). Antioxidant capacity of 1-deoxy-D-
erythro-hexo-2,3-diulose and D-arabino-hexo-2-ulose. Journal of Agricultural
and Food Chemistry, 62, 2837e2844.
Knorr, D., Fro hling, A., Ja ger, H., Reineke, K., Schlter, O., & Scho
ssler, K. (2011).
Emerging technologies in food processing. Annual Review of Food Science and
Technology, 2, 203e235.
Koubaa, M., Barba, F. B., Grimi, N., Mhemdi, H., Koubaa, W., Boussetta, N., et al.
(2016). Recovery of colorants from red prickly pear peels and pulps enhanced
by pulsed electric eld and ultrasound. Innovative Food Science & Emerging
Technologies, 37, 336e344. http://dx.doi.org/10.1016/j.ifset.2016.04.015.
Loginova, K. V., Lebovka, N. I., & Vorobiev, E. (2011). Pulsed electric eld assisted
aqueous extraction of colorants from red beet. Journal of Food Engineering, 106,
127e133.
rez-Santana, L. (1999). Preliminary studies of analgesic and
Loro, J. F., del Rio, I., & Pe
anti-inammatory properties of Opuntia dillenii aqueous extract. Journal of
Ethnopharmacology, 67, 213e218.
Min, S., Jin, Z. T., Min, S. K., Yeom, H., & Zhang, H. Q. (2003). Commercial-scale pulsed
electric eld processing of orange juice. Journal of Food Science, 68, 1265e1271.
Moussa-Ayoub, T. E., Abd El-Hady, E. A., Omran, H. T., El-Samahy, S. K., Kroh, L. W., &
Rohn, S. (2014). Inuence of cultivar and origin on the avonol prole of fruits
and cladodes from cactus Opuntia cus-indica. Food Research International, 64,
864e872.
Moussa-Ayoub, T. E., El-Samahy, S. K., Rohn, S., & Kroh, L. W. (2011). Flavonols,
betacyanins content and antioxidant activity of cactus Opuntia macrorhiza
fruits. Food Research International, 44, 2169e2174.
Moussa-Ayoub, T. E., Jaeger, H., Youssef, K., Knorr, D., El-Samahy, S., Kroh, L. W., et al.
542 T.E. Moussa-Ayoub et al. / LWT - Food Science and Technology 79 (2017) 534e542
(2016). Technological characteristics and selected bioactive compounds of
Opuntia dillenii cactus fruit juice following the impact of pulsed electric eld
pre-treatment. Food Chemistry, 210, 249e261.
thlings, U., Murphy, S. P., Wilkens, L. R., Boeing, H., Schulze, M. B., Bueno-de-
No
Mesquita, H. B., Kolonel, L. N. (2008). A food pattern that is predictive of
avonol intake and risk of pancreatic cancer. The American Journal of Clinical
Nutrition, 88, 1653e1662.
Odriozola-Serrano, I., Soliva-Fortuny, R., & Martn-Belloso, O. (2008). Phenolic acids,
avonoids, vitamin C and antioxidant capacity of strawberry juices processed
by high-intensity pulsed electric elds or heat treatments. European Food
Research and Technology, 228, 239e248.
Patras, A., Brunton, N. P., Da Pieve, S., & Butler, F. (2009). Impact of high pressure
processing on total antioxidant activity, phenolic, ascorbic acid, anthocyanin
content, and colour of strawberry and blackberry pure es. Innovative Food Sci-
ence & Emerging Technologies, 10, 308e313.
Patterson, M. F. (2005). A review: Microbiology of pressure-treated foods. Journal of
Applied Microbiology, 98, 1400e1409.
Ruan-Henares, J. A., & Morales, F. J. (2007). Functional properties of melanoidins:
In vitro antioxidant, antimicrobial and antihypertensive activities. Food Research
International, 40, 995e1002.
S
anchez-Moreno, C., Plaza, L., Elez-Martnez, P., De Ancos, B., Martn-Belloso, O., &
Cano, M. P. (2005). Impact of high pressure and pulsed electric elds on
bioactive compounds and antioxidant activity of orange juice in comparison
with traditional thermal processing. Journal of Agricultural and Food Chemistry,
53, 4403e4409.
Smuda, M., & Glomb, M. A. (2013). Maillard degradation pathways of vitamin C.
Angewandte Chemie International Edition, 52, 4887e4891.
p, S., Mathys, A., Heinz, V., & Knorr, D. (2006). Review: Potential of high hy-
To
drostatic pressure and pulsed electric elds for energy efcient and environ-
mentally friendly food processing. Food Reviews International, 22, 405e423.
Yeom, H. W., Streaker, C. B., Zhang, Q. H., & Min, D. B. (2000). Effects of pulsed
electric elds on the activities of microorganisms and pectin methyl esterase in
orange juice. Journal of Food Science, 65, 1359e1363.
Yilmaz, Y., & Toledo, R. (2005). Antioxidant activity of water-soluble Maillard re-
action products. Food Chemistry, 93, 273e278.
Zhang, N., Pei, F., Wei, H., Zhang, T., Yang, C., Ma, G., et al. (2011). Isorhamnetin
protects rat ventricular myocytes from ischemia and reperfusion injury.
Experimental and Toxicologic Pathology, 63, 33e38.
Zou, H., Lin, T., Bi, X., Zhao, L., Wang, Y., & Liao, X. (2016). Comparison of high hy-
drostatic pressure, high-pressure carbon dioxide and high-temperature short-
time processing on quality of mulberry juice. Food and Bioprocess Technology, 9,
217e231.
Zulueta, A., Barba, F. J., Esteve, M. J., & Frgola, A. (2013). Changes in quality and
nutritional parameters during refrigerated storage of an orange juice-milk
beverage treated by equivalent thermal and non-thermal processes for mild
pasteurization. Food and Bioprocess Technology, 6, 2018e2030.