Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11

Contents lists available at ScienceDirect

Chemical Engineering & Processing: Process Intensification

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

Microwave-assisted extraction of phenolics from pomegranate peels: T

Optimization, kinetics, and comparison with ultrasounds extraction
⁎
Kyriakos Kaderides, Lygeri Papaoikonomou, Melania Serafim, Athanasia M. Goula
Department of Food Science and Technology, School of Agriculture, Forestry and Natural Environment, Aristotle University, 541 24, Thessaloniki, Greece

A R T I C LE I N FO A B S T R A C T

Keywords: Recently, there has been much interest in the phenolics of pomegranate peels, because of their health benefits,
Antioxidant activity such as antioxidant activity. Thus, the objective of this work is to optimize an efficient alternative and “green”
Microwave extraction technique for the extraction of phenolics from pomegranate peels. Microwave-assisted method was found to
Phenolics increase extraction yield, but mainly to shorten the treatment time by over 60 times compared to conventional
Pomegranate peel
extraction methods. The effects of solvent type, solvent/solid ratio, and microwave power on the yield of
Punicalagin
Ultrasound extraction
phenolics extraction were studied. The optimum operating conditions were found to be: solvent type, 50%
aqueous ethanol; solvent/solid ratio, 60/1 mL/g; power, 600 W. The efficiency of the proposed extraction pro-
tocol was compared with that of the ultrasound-assisted extraction, another “green” extraction approach, studied
in our previous work. The microwaves method had a yield about 1.7 times higher obtained in a shorter process
time (4 min) in comparison to ultrasound-assisted extraction (10 min). These differences were attributed to the
intense cell destruction of microwave-treated plant material observed by SEM analysis. The obtained extract
presented a high antioxidant activity (radical scavenging activity of 94.91%), due to the high content of puni-
calagin (143.64 mg/g dry matter) measured by HPLC analysis, that was equivalent to that of the ultrasounds
extract (radical scavenging activity, 94.77%; punicalagin content, 138.8 mg/g dry matter).

1. Introduction (e.g., as ingredient in functional foods). For example, PPE afforded

Pomegranate fruit (Punica granatum L.) is composed of three parts:
the seeds, the juice, and the peels. Pomegranate peels constitute ap-
proximately 40% of the whole fruit and are rich in phenolics com-
pounds such as ellagitannins, punicalagin, and punicalin [1]. Pome-
granate peels have been traditionally valorized as animal feed and only
small quantities are used in medicine. However, most of them are di-
rectly disposed as waste.
Several studies have reported the in vitro bioactivity of pome-
granate peel extracts including antioxidant, antitumor, anti-in-
flammatory, and antiproliferative properties. Kanatt et al. [2] in-
vestigated the antioxidant and antimicrobial potential of pomegranate
peel extract (PPE) and concluded that the efficacy of PPE in scavenging
hydroxyl and superoxide anion radical was very high. In addition, the
extract had good reducing power and iron chelation capacity and
showed good antimicrobial activity against Staphylococcus aureus and
Bacillus cereus having minimum inhibitory concentration of 0.01%.
Pseudomonas could be inhibited at a higher concentration of 0.1% while
it was ineffective against Escherichia coli and S. typhimurium. Thus, PPE
could potentially be included in several industries due to its versatility
enhanced antioxidant activity by inhibiting lipid oxidation in cooked
chicken patties [3], whereas Kanatt et al. [2] found that addition of PPE
to popular chicken meat products enhanced their shelf life by 2–3
weeks during chilled storage. Pomegranate peel extract at concentra-
tions of 800–850 ppm in sunflower oil [4] and 200–1000 ppm in fish oil
[5] had stabilization efficiency comparable to conventional synthetic
antioxidants, i.e. BHT at its legal limit, whereas Kumudavally et al. [6]
and Devatkal et al. [7] concluded that PPE significantly increases the
stability of beef and goat meat products against lipid peroxidation.
According to Cam et al. [8], incorporation of pomegranate peel phe-
nolics into ice cream resulted in sharp improvements in antioxidant and
antidiabetic activities as well as the phenolic content of ice creams.
Addition of PPE to jams [9], juices and wines [10] increased their
phenolic, flavonoids, and thiol concentration with a significant im-
provement of the free radical scavenging and product stability features.
Recently, Kaderides et al. [11] incorporated pomegranate peel extract
in hazelnut paste and reported an inhibition of lipid oxidation with
reduced formation of peroxides.
Pomegranate peel phenolics can be extracted with conventional
systems such as Soxhlet and maceration. Shortcomings of existing

⁎
Corresponding author.
E-mail address: athgou@agro.auth.gr (A.M. Goula).

https://doi.org/10.1016/j.cep.2019.01.006
Received 22 November 2018; Received in revised form 3 January 2019; Accepted 10 January 2019
Available online 24 January 2019
0255-2701/ © 2019 Elsevier B.V. All rights reserved.
K. Kaderides et al.
conventional extraction technologies, like increased consumption of
energy, high extraction time, possible degradation of bioactive com-
pounds, and high consumption of harmful chemicals, have forced the
food and chemical industries to find new separation “green’’ techni-
ques, which typically use less solvent and energy, such as ultrasound-
assisted extraction [11–14] and microwave-assisted extraction [15–17].
Microwave assisted extraction (MAE) is one of the most employed
alternative extraction methods. Among the advantages reported for
MAE are its lower extraction times and solvent consumption. The
process acceleration and the high extraction yield may be the result of a
synergistic combination of two transport phenomena: heat and mass
gradients working in the same direction [18]. According to Zheng et al.
[19], MAE is characterized by disruption of cell structure caused by the
penetrating volume heating from microwave irradiation. The diffusion
capacities of target components may be improved by the increment of
solvent temperature under microwave heating. So far, microwave
treatment has been widely employed to extract phenolics from different
plant materials including citrus peels [20], peanut skins [21], grape
seeds [22], tomatoes [23], blueberries [19], spent filter coffee [24],
chokeberries [25], and walnut leaves [26].
Therefore, MAE is a potential method for the extraction of phenolics
from pomegranate peels. According to Zheng et al. [19], the optimi-
zation of extraction parameters is helpful to systematically understand
the interaction between independent factors and obtain the optimal
parameter combination for optimum extraction. In addition, the MAE
efficiency depends on several variables which may not be generalized
for all plant materials due to the diverse nature of existing bioactive
phytochemicals, being necessary to select and optimize the processing
conditions as a function of the used matrix and taking into account the
desired responses [27]. To our best knowledge, as far as pomegranate
peel is concerned, only Zheng et al. [16] and Huang et al. [17] em-
ployed MAE to extract phenolic compounds with water and flavonoids
with aqueous ethanol, respectively.
According to Shirsath et al. [28], mathematical modelling can help
in the design, optimization, and control of the extraction process, as
well as provide useful information for scale up of the equipment. The
typical kinetic models of extraction include unsteady diffusion, Fick’s
law of diffusion, film theory, and empirical models. However, no in-
formation is available for kinetic modeling of microwave-assisted ex-
traction of phenolics from pomegranate peels.
Therefore, the objective of this study is to develop an efficient al-
ternative and “green” technique for the extraction of phenolics from
pomegranate peels. The operating parameters are optimized and the
mechanism of the enhanced extraction is discussed by observing cell
destruction of plant material. The efficiency of the proposed extraction
protocol is compared with that of another “green” extraction approach.
The content of the major phenolic compounds and the antioxidant ac-
tivity are also evaluated. In addition, a mathematical model to describe
the kinetic mechanisms of polyphenols extraction is developed,
whereas the dependence of the best descriptive model constants on
extraction variables is expressed by an appropriate type of model. These
results would offer scientific reference for promoting the valorization of
pomegranate peels.
2. Materials and methods
2.1. Standards and reagents
HPLC-grade acetonitrile was from Fisher Scientific (Lisbon,
Portugal). The phenolic compound standards were from Sigma-Aldrich
(St. Louis, MO, USA). All the other chemicals were of analytical grade.
2.2. Pomegranate peels
Pomegranate peels as by-product of fruit juice industry were pro-
vided from a local producer (Rodi Hellas, Greece). The peels were dried
2
Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11
at 40 °C for 48 h and kept at −30 °C until use. The peels were ground in
a laboratory mill (Type A10, Janke and Kunkel, IKA Labortechnik,
Germany) immediately prior to extraction. The particle size distribution
of the milled pomace was determined by a sieve shaker (Retax, Labor
Siebmaschine, Type LS10, Nr 4082, Germany). The particle mean dia-
meter was about 0.1 mm.
2.3. Microwave-assisted extraction
Microwave-assisted extraction was performed with a Multiwave
closed microwave system (B30MC030A, AntonPaar, Graz, Austria),
equipped with standard configuration with rotor for 6 sample vessels
type H (50 mL quartz tubes; operating pressure 75 bars). A sample of
powdered pomegranate peels was mixed with 20 mL solvent to produce
different solvent/peel ratios. The extracts were collected at 0.5, 1, 2, 3,
4, 5, 10, and 15 min. The resulting extracts were evaporated using a
rotary evaporator (Rotovapor R114, Waterbath B480, Büchi, Flawil,
Switzerland). The variation of extraction yield during the extraction
process was studied with various (i) microwave powers (P) (100, 173,
350, 527, and 600 W) and (ii) solvent/peels ratios (L/S) (10.0, 17.3,
35.0, 52.7, and 60.0 mL/g). A central composite design was applied to
determine the effects of the above parameters on extraction yield.
Thirteen experimental runs were carried out and each one was con-
ducted with five different solvents: water, 50 and 70% aqueous ethanol,
and 50 and 70% aqueous methanol.
Extraction yield, Y (%), was defined as the percent ratio of the total
weight of gallic acid equivalents extracted to the dry peels sample
weight.
2.4. Determination of total phenolics content and antioxidant activity
The total phenolic content of extracts was determined according to
the Folin-Ciocalteu method [29]. Briefly, 0.2 mL of the extract was
added to a 25 mL volumetric flask and additional distilled water was
added to make a final volume of 10 mL. A reagent blank was prepared
using distilled water. Folin-Ciocalteu phenol reagent (0.5 mL) was
added to the mixture and shaken vigorously. After 5 min, 5 mL of 5%
Na2CO3 solution was added with mixing. The solution was immediately
diluted to 25 mL with distilled water, mixed thoroughly and then al-
lowed to stand for 90 min. The absorbance was measured at 750 nm
versus the prepared blank. The total phenolic content of the sample was
expressed as gallic acid equivalents (GAE) mg/mL.
Antioxidant activity was determined using DPPH, which is a stable
free radical and is widely used to assess the radical scavenging activity
of antioxidant components. This method is based on the reduction of
DPPH in methanol solution in the presence of a hydrogen donating
antioxidant due to the formation of the non radical form DPPH-H [30].
The transformation results in a color change from purple to yellow,
which is measured spectrophotometrically at 517 nm. The reaction
mixture (3.0 ml) consists of 1.0 ml of DPPH in methanol (0.3 mM),
1.0 ml of extract, and 1.0 ml of methanol. The percentage of inhibition
can be calculated using the Eq. (1).
Inhibition (%) = ((A0–A1)/A0) × 100 (1)
where A0 is the absorbance of the control and A1 is the absorbance of
the sample.
2.5. Analysis of phenolic compounds
The samples were analyzed using a HPLC system equipped with two
Marathon IV pumps (Spark NL., The Netherlands), a Timberline TL-50
temperature controller (Indiana, U.S.A.), a UV–vis (UV 6000 LP de-
tector -Spectra SYSTEM, Finningan Mat, Thermo Separation Products,
San Jose, U.S.A) and a hypersil C18 reversed-phase column
(250 × 4.6 mm I.D., 5 μm particle size). The mobile phase was a
K. Kaderides et al.
mixture of water/acetic acid (95:5) (A) and acetonitrile (B). Samples
were analyzed using a gradient program developed by Kaderides et al.
[31]. Analyses were conducted at constant temperature of 30 °C using a
flow rate of 1 mL/min and a sample injection volume of 10 μL. Double
online detection was carried with a diode array detector operating at
254 and 280 nm as preferred wavelengths and the target phenolic
compounds were identified according to their UV spectra and retention
time. For the quantitative analysis, a baseline to valley integration with
baseline projection mode was used to calculate peak areas and external
standards were used for quantification. The results were expressed in
mg per g of dry weight.
2.6. Scanning electron micrographs (SEM)
A Quanta-200 environmental scanning electron microscope system
(FEI Company, USA) was used to study the structural changes in po-
megranate peels texture after the extraction process. The samples were
fixed on a specimen holder with an aluminum tape and sputtered with a
thin layer of gold prior to examination under high vacuum condition at
an accelerating voltage of 15 kV at 2000× magnification level.
2.7. Kinetic models
In the present study, three different mathematical approaches were
selected for the kinetic modelling of extraction of polyphenols, namely,
the Peleg’s model, the first-order kinetic model, and the second-order
kinetic model. These three models were compared by fitting the ex-
perimental data obtained from extraction into each model.
2.7.1. Peleg’s model
Peleg’s equation [Eq. (2)] is a non-exponential empirical equation
proposed as [32]:
t
Ct =
K1 + K2⋅t (2)
where Ct is the concentration of polyphenols in the extract (mg/mL) at
time t (min), K1 is Peleg’s rate constant (min mL/mg), and K2 is Peleg’s
capacity constant (mL/mg). Peleg’s K1 parameter refers to extraction
rate (B0, mg/min mL) at the very beginning (t = t0) according to Eq.
(3):
1 3. Results and discussion
B0 =
K1 (3)
Peleg’s capacity constant K2 refers to the maximum extraction yield,
that is, the equilibrium concentration of total extracted analyte when t
→ ∞ (Ceq, mg/mL), as indicated in Eq. (4).
1 (Fig. 1). Thus, the efficient extraction period for achieving maximum
Ceq =
K2 (4)
2.7.2. First-order kinetic model
According to Poojary and Passamonti [32], when we plot the con-
centration of extracted polyphenols in the matrix against time, the
curve which describes the extraction kinetics usually shows an ex-
ponential decay. The most usual form of the first-order kinetic model is
presented in Eq. (5).
Ct = Ceq (1 − e−KA⋅ t )
(5)
where KA is the overall extraction rate constant (1/min).
2.7.3. Second order kinetic model
The solid-liquid extraction process can be considered as the reverse
of an adsorption operation, therefore the bases of the adsorption kinetic
equations can be applied to solid-liquid extraction and the second-order
law was found to give the best fits for the extraction rate [33]. The
general second-order model can be written as [13]:
3
Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11
dCt
= KB⋅(Ceq − Ct )2
dt (6)
where KB is the second-order extraction rate constant (mL/mg min).
The integrated rate law for a second-order extraction under the
boundary conditions t = 0 to t and Ct = 0 to Ct, can be written as an Eq.
(7):
2
KB⋅t⋅Ceq
Ct =
1 + KB⋅t⋅Ceq (7)
where h is the initial extraction rate (mg/mL min) when t approaches 0:
2
h = KB⋅Ceq (8)
2.8. Statistical analysis
The parameters of the kinetic models were estimated by nonlinear
regression. To evaluate the goodness of fit of each model, two criteria
were used: the coefficient of determination, R2, which is the relative
variance explained by the model with respect to the total variance, and
the standard error of the estimate, SEE (Eq. (9)), which is a measure of
the accuracy of predictions.
∑ (Ct , exp − Ct , pred )2
SEE =
N (9)
where Ct,exp is the experimental value of polyphenols concentration in
the extract at time t, Ct,pred is the predicted concentration, and N is the
number of observations.
The effect of process variables on extraction yield and on kinetic
parameters were analyzed using the statistical software MINITAB
(Release 13.32). To identify the significance of the effects and inter-
actions between them, analysis of variance (ANOVA) was performed. A
p value less than 0.05 was considered to be statistically significant.
Regression analysis was used to fit full second order polynomials, re-
duced second order polynomials, and linear models to the data of each
of the variables evaluated (response variables). F values for all reduced
and linear models with an R2 > 0.70 were calculated to determine if the
models could be used in place of full second order polynomials to
predict the response of a variable to the independent variables.
3.1. Effect of process variables on microwave-assisted extraction yield
The extraction yield was time-dependent and increased with ex-
tended irritation times from 0.5 to 4 min and then slightly decreased
yield of pomegranate peel phenolics was about 4 min. According to
Veggi, Martinez, and Meireles [18], the extraction process takes place
in three different steps: an equilibrium phase where the phenomena of
solubilization and partition intervene, in which the substrate is re-
moved from the outer surface of the particle at an approximately con-
stant velocity. Then, this stage is followed by an intermediary transition
phase to diffusion, in which mass transfer by convection and diffusion
prevails. In the last phase, the extraction rate is low and the solute must
overcome the interactions that bind it to the matrix and diffuse into the
extracting solvent. During microwave-assisted extraction of pome-
granate peel phenolics, the absorption of microwave energy in the ex-
traction system promoted the thermal accumulation of the extraction
solution resulting in the dissolution of phenolics into the solution until
4 min [34]. The same behavior was observed by Xie et al. [35] and Wu
et al. [36] for extraction of flavonoids from Cyclocarya paliurus Iljins-
kaja leaves and phenolics from potatoes, respectively. Huang et al. [17]
and Alara et al. [37] reported an initially increase in extraction yield
with increasing irritation time reaching a maximum value, during ex-
traction of phenolics from pomegranate peels and Vernonia amygdalina
K. Kaderides et al.
Fig. 1. Main effects plot illustrating the effect of extraction time (t, min), microwave power (P, W), and solvent-to-peels ratio (L/S, mL/g) on the extraction yield (Y,
mg GAE/g dry peel).
leaves. However, in all these studies the excessive time exposure in the
microwave field decreased the extraction yield due to a possible de-
gradation of phenolic compounds [37].
The most important factor that affects MAE is solvent selection that
depends on the solubility of the compounds of interest, solvent pene-
tration and its interaction with the sample matrix and its dielectric
constant, and the mass transfer kinetics of the process [18]. In this
work, the ability of different solvents in extracting phenolic compounds
from pomegranate peels was compared. Fig. 2 is the main effects plot
for solvent type, which was found to have a significant effect
(p < 0.05) on total phenolic yield. Extraction yield increased in the
following order of solvents: 70% methanol < 50% methanol <
water < 70% ethanol < 50% ethanol.
Ethanol-water mixtures seem to be the most suitable solvents for the
extraction. Kaderides, Goula, and Adamopoloulos [11] reported that
the extraction yield increases with an increase in solvent polarity and a
decrease in viscosity and the higher the molecular weight of the solvent,
the lower the polarity, which allows other substances of about the same
molecular weight to be easily extracted. This can be correlated to “like
dissolve like” or “polarity versus polarity” principle. Ethanol, methanol,
and water have a relative polarity of 0.654, 0.762, and 1.0, respectively
[38]. Methanol is considered more efficient solvent for extraction of
antioxidant compounds than ethanol even if their polarities are similar.
This may be due to the low solvation provided by ethanol, probably
because of the presence of the ethyl radical that is longer than the
Fig. 2. Main effect of solvent type on the extraction yield (Y, mg GAE/g dry
peel) obtained at 4 min.
4
Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11
methyl radical present in methanol, resulting in a lower solvation of
antioxidant molecules [39]. However, in the case of microwave ex-
traction, the loss tangent is the ratio of the dielectric loss to the di-
electric constant, a dielectric parameter which can be used as an index
of the efficiency with which electromagnetic radiation is converted to
heat [40]. The loss tangent of ethanol and methanol at 2.45 GHz and
30 °C is 0.898 and 0.522, respectively [41], and of water at 2.45 GHz
and 25 °C is 0.123 [42]. This means that water is considered a medium
microwave absorber, methanol is a good absorber, while ethanol is
considered a strong microwave absorber. Hence, ethanol has higher
ability than methanol to absorb microwaves and convert them to heat,
promoting both the rapid rupture of the plant material and the in-
creased diffusion of the targeted compounds [40].
In addition, it was found that a binary solvent is more efficient for
extraction of phenolics than a mono-solvent one [43]. Addition of water
to a solvent usually creates a more polar medium, which facilitates the
extraction of polyphenols as suggested by Spigno et al. [44]. By in-
creasing the proportion of water, the polarity of solvent also increases
and the system is able to extract phenolic substances from both ends of
the polarity (highest polarity substances and low polarity substances),
as well as those of moderate polarity. Several researchers reported that
an increase of ethanol concentration in a binary ethanol-water solvent
results in the addition of the polarity molecular to close that in phenolic
compounds. Thus, the phenolics solubility may be improved with the
ethanol concentration based on the “polarity versus polarity” principle.
Zheng et al. [19] explained that ethanol plays a major role to rupture
the hydrogen bonds and hydrophobic bonds existing between pheno-
lics–proteins and phenolics–cellulose in the water–ethanol system.
However, in the present work, 50% ethanol was found more efficient
than 70% ethanol. This finding may be associated to the fact that both
the dielectric constant and dielectric loss factor of water are higher than
those of ethanol and under the same irradiated intensity of microwave
output power, water may absorb more thermal energy than ethanol to
drive target component to diffuse [19]. A similar trend was reported by
Zheng et al. [19], who concluded that the more appropriate ethanol
concentration for microwave-assisted extraction of anthocyanin from
blueberry is around 60%, and Taha et al. [45], who studied the effect of
ethanol concentration (ranged from 50 to 80%) on the total content of
phenolic compounds in sunflower meal extracts, found that 60%
ethanol was the most efficient.
Figs. 1 and 3 presents the effects of microwave power (P) and sol-
vent/peel ratio (L/S) on extraction yield. As the liquid-to-solid ratio
K. Kaderides et al.
Fig. 3. Contour plots illustrating the effect of microwave power (P, W) and
solvent-to-peels ratio (L/S, mL/g) on the extraction yield (Y, mg GAE/g dry
peel) obtained at 4 min using 50% aqueous ethanol.
increases from 10.0/1 to 52.7/1 mL/g, the extraction yield is sig-
nificantly enhanced. In general, a higher ratio of solvent to raw material
results in a larger concentration gradient during the diffusion from the
solid into the solution and a higher extraction yield. In addition, the
increase of solvent/solid ratio leads to excessive swelling of the plant
material increasing the contact surface area between the material and
the solvent [34]. However, with further increase in liquid/solid ratio, a
decrease in yield was observed. Shirsath et al. [28], who studied the
extraction of curcumin from Curcuma amada, concluded that if the so-
lution is very dilute, an increased quantity of solvent would not lead to
a sufficient increase in the concentration gradient and the increase in
extraction yield would be limited. A large solvent volume would de-
crease the microwave adsorption of material, since more energy is
needed to be absorbed by the solvent [17]. According to Alara et al.
[37], a very high solvent/solid ratio may result to insufficient energy in
facilitating the cell wall breakage for effective leaching out of the
phenolic compounds. A similar observation was reported by Galan et al.
[40] and Huang et al. [17], who found the maximum extraction yield of
phenolics from Citrus sinensis peels and pomegranate peels at a liquid/
solid ratio of 30/1 and 40/1 mL/g, respectively. Also, using a large
amount of solvent is not considered to be cost-effective due to high
operating cost of solvents and energy consumption required during the
subsequent separation [28]. However, Zheng et al. [16] reported that
the effect of liquid/solid ratio on recovery of phenolics from pome-
granate peels was not significant. Thus, the optimum value of solvent/
solid ratio is specific to the system under investigation and needs to be
established experimentally.
As far as the effect of microwave power (P) is concerned, the ex-
traction yield generally increased with increasing the power from 173
to 600 W, whereas an opposite trend was observed when power in-
creased from 100 to 173 W. It must be underlined that the extraction
temperature, which increases with microwave power, could not be
adjusted in the microwave system used in the present work. The first
effect may be attributed to the fact that increasing the microwave
power, more electromagnetic energy was transferred on the molecules
by ionic conduction and dipole rotations resulting in rapidly heating of
the extraction system [46]. At higher temperatures, the solubility and
diffusion coefficients of the phenolic compounds to be extracted in-
crease and the viscosity of the solvent decreases, thus facilitating its
passage through the solid substrate mass. Moreover, the microwave
irradiation accelerates cell rupture by sudden internal pressure increase
inside the cells of plant sample, which promotes the destruction of
sample surface and in turns the exudation of phenolic compounds
within the plant cells into the surrounding solvent [47]. According to
Liew et al. [48], some researchers prefer the approach of low micro-
wave power with longer extraction time as high power may reduce
purity. However, this approach may cause overheating and reduce the
5
Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11
nutrient content of the extracted compounds. However, at power lower
than 173 W, a decrease of extraction yield was observed with increasing
the power. According to Maran et al. [49], this effect may be due to the
fact that high irradiation power offered redundant energy to the solvent
and material drastically disturbing molecular interactions.
According to the ANOVA analysis of the optimization study, the
liquid/solid ratio and its square, both with a probability value of 0.001,
influenced most significantly the extraction yield. The regression
coefficients were calculated and the data was fitted to a second-order
polynomial equation. The response extraction yield (Y) can be ex-
pressed in terms of the regression Eq. (10):
Y = 140.62 − 0.008⋅P + 0.996⋅L/ S + 0.002⋅(L/ S )2 (10)
2
The coefficient of determination, R , was 0.912, indicating that 91.2%
of the total variability in the response could be explained by the specific
model, whereas an adjusted R2 value of 0.891 revealed that there was a
good agreement between experimental and predicted values of ex-
traction yield. The model F-value was 3.15, corresponding to a p-value
of less than 0.05, which, along with the p-value for lack of fit (0.188),
indicated that the model fits the experimental data accurately.
The desirability profile for optimum yield indicated that the max-
imum desirability level of 1.0 (on a scale of 0–1) can be achieved with a
microwave power of 600 W, a solvent/solid ratio of 60/1 mL/g, and
50% aqueous ethanol as solvent at an extraction time of 4 min. Under
these optimized conditions, the predicted value for extraction yield was
202.8 mg GAE/g dry peels, whereas the observed experimental value
was found 199.4 mg GAE/g dry peel. A similar value was reported by
Zheng et al. [16], who found the optimum extraction yield of total
phenolics from pomegranate peel at 214.5 mg GAE/g dry peel.
3.2. Microwave-assisted extraction kinetics
Different kinetic models were employed to fit with experimental
data. The highest values of R2 and lowest values of SEE were chosen for
goodness of fit. Table 1 presents the R2 and SEE values for the kinetic
models. Among these, the Peleg’s model fits the best with experimental
values showing the highest average values of R2 (0.924–0.991) and the
lowest average values of SEE (0.889–1.322). The applicability of Peleg’s
equation on food materials has been demonstrated extensively [32].
The kinetic parameters of the Peleg’s model at different extraction
conditions are presented in Fig. 4. It should be noted that a lower K1
value in Eq. (2) implies a faster rate of the process, whilst the lower K2
value indicates maximum yield [50]. According to Ji et al. [51], the
extraction rate is controlled not only by mass transfer resistance in the
liquid film around solute particles, but also by intra-particle diffusion;
hence when the mass transfer resistance in the liquid film is reduced,
the extraction rate is controlled only by intra-particle diffusion. As it
can be seen, the kinetic parameters generally increased with an increase
in liquid-to-solid ratio. Higher liquid to solid ratio ensures homogenous
mixing and allows penetration of solvent into deep interior parts [52].
However, the capacity and the rate decreased with an increase in L/S.
This effect may be attributed to the fact that for microwave irradiation
the temperature rise is higher at lower solvent-to-feed ratio owing to
the lower volume of solvent [53]. As far as microwave power is con-
cerned, power provides localized heating in the sample, which acts as a
driving force for MAE to destroy the plant matrix so that the solute can
diffuse out and dissolve in the solvent, thus increasing the rate constant.
However, for high solvent-to-solid ratios, rapid rupture of the cell wall
can take place at a higher temperature when using higher power, and as
a result impurities can also be leached out into the solvent together with
the desired solute [18]. Thus, the constant K1 presents a maximum at a
microwave power around 400 W. As it can be seen in Fig. 4, the ex-
traction capacity slightly decreased with an increase in P. This finding
may be associated with a possible degradation of thermally sensitive
compounds at high microwave powers.
Fig. 5 presents the comparison of predicted and experimental values
K. Kaderides et al. Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11

Table 1
Statistical parameters of the kinetic models for microwave-assisted extraction of polyphenols from pomegranate peels using 50% aqueous ethanol.
A/A Microwave power (P, W) Solvent/peel ratio (L/S, mL/g) Kinetic model

Peleg’s model First-order kinetic model Second-order kinetic model

R2 SEE R2 SEE R2 SEE

1 350 10.0 0.972 1.322 0.971 1.361 0.965 1.481

2 527 17.3 0.991 0.498 0.987 0.606 0.973 0.885
3 173 17.3 0.965 0.839 0.958 0.919 0.911 1.347
4 100 35.0 0.964 0.478 0.935 0.640 0.907 0.769
5 600 35.0 0.976 0.468 0.974 0.486 0.938 0.744
6 350 35.0 0.978 0.401 0.952 0.584 0.941 0.650
7 350 35.0 0.972 0.442 0.938 0.651 0.908 0.795
8 350 35.0 0.982 0.376 0.952 0.623 0.922 0.793
9 350 60.0 0.930 0.539 0.962 0.399 0.509 1.431
10 350 35.0 0.953 0.630 0.920 0.818 0.912 0.861
11 527 52.7 0.977 0.289 0.978 0.286 0.948 0.433
12 350 35.0 0.979 0.438 0.953 0.653 0.893 0.983
13 173 52.7 0.924 0.523 0.882 0.652 0.832 0.777
Average 0.966 0.557 0.951 0.668 0.889 0.919

Fig. 5. Experimental and predicted by the Peleg’s model values of polyphenols

concentration in the extract (C, mg/mL) at a microwave power of 350 W, a
solvent-to-peels ratio of 35.0/1 mL/g, and 50% aqueous ethanol as solvent.
Prediction Band is the region where 95% of the experimental data points are
expected to be and Confidence Band is the region where 95% of the regression
lines are expected to be.

Qu et al. [54], who studied the extraction of antioxidants from

Fig. 4. Contour plots illustrating the effect of microwave power (P, W) and
solvent-to-peels ratio (L/S, mL/g) on the kinetic parameters of the Peleg’s
model (K1 in min mL/mg and K2 in mL/mg).
of polyphenols concentration in the extract for the Peleg’s model at a
microwave power of 350 W, a liquid-to-solid ratio of 35.0/1 mL/g, and
50% aqueous ethanol as solvent. The figure presents also the confidence
(CB) and prediction (PB) bands for the polyphenols concentration at the
solvent phase against time. It is noted that similar trends were observed
for all experiments. The PB is the region where 95% of the experimental
data points are expected to be and it is observed that all obtained ob-
servations fall within. Likewise, the CB is the region where 95% of the
regression lines are expected to be.
pomegranate marc, fitted the relationships between kinetic parameters
and processing factors (particle size, water/sample ratio, and tem-
perature) by linear, power or second-order polynomial functions.
However, they developed a different model for each factor. In this
work, multiple regression analysis was used to develop equations pre-
dicting the effect of all extraction factors simultaneously. The effect of
extraction variables on Peleg’s model constants can be expressed by
Eqs. (11) and (12).
K1 (×10−3) = 4.006 + 0.445⋅P − 5.469⋅L/ S + 0.153⋅L/ S 2
− 0.003⋅P⋅L/ S (R20.911) (11)
K2 (×10−3) = 42.467 + 0.166⋅P − 3.641⋅L/ S + 0.018⋅L/ S 2
+ 0.001⋅P⋅L/ S (R2 = 0.962) (12)
Substituting Eqs. (11) and (12) into Eq. (2), a kinetic model for
predicting polyphenols extraction from pomegranate peel can be ob-
tained. Even though the empirical models (11)–(12) cannot account for
the phenomena governing extraction processes, they could be used to
determine the effects of microwave power and liquid-to-solid ratio on

6
K. Kaderides et al.
Fig. 6. HPLC chromatogram of pomegranate peel extract at the optimum conditions of microwave- and ultrasound-assisted extraction (254 nm).
the phenolics extraction yield [33].
3.3. HPLC analysis
Polyphenol qualitative and quantitative determination of pome-
granate peel extract was performed by HPLC analysis. The identified
phenolic components were quantified with respect to reference sub-
stances to compare the content of the major individual phenolic com-
ponents in pomegranate peel extract. Generally, limited research has
7
Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11
been achieved with regard to the quantification of phenolic compounds
in pomegranate peel extracts using HPLC; furthermore, it is difficult to
compare results from different studies since different extraction
methods were used. In the present work, punicalagin was determined as
predominant in the extract at the optimum conditions of microwave-
assisted extraction (Fig. 6). This observation is similar to that reported
by other researchers. Aqil et al. [55] and Gullon et al. [56] reported that
punicalagin is the main bioactive compound present in pomegranate
peel. Similarly, Yan et al. [57] found that among the phenolic
K. Kaderides et al.
compounds of pomegranate peel, punicalagin present as a mixture of
two reversible anomers (a and b) was the dominant one, accounting for
more than 90% of the phenolics in all of the tested cultivars (range,
17.8–114.5 mg/g). The alpha and beta forms of pomegranate punica-
lagin are polyphenolic hydrolysable tannins and isomers of 2,3-(S)-
hexahydroxydiphenoyl-4,6-(S,S)-gallagyl- D-glucose [58].
Punicalagin content (a + b) of the pomegranate peel extract at the
optimum conditions of microwave-assisted extraction was determined
as 143.64 mg/g on dry matter basis. This punicalagin content is higher
than the values reported by Lu et al. [59] (39.8–121.5 mg/g dry weight)
for pomegranate peel extracts obtained by UAE for 30 min in 40%
aqueous ethanol. Gullon et al. [56] applying ultrasound-assisted ex-
traction with 50% aqueous ethanol determined a lower punicalagin
content of 16.67 mg/g dry weight. On the contrary, Kharchoufi et al.
[60] using the normal stirring method, reported much higher values of
punicalagin, respectively 193.78 mg/g for a methanolic extract and
245.47 mg/g for an aqueous one. Such variations could be due to the
extraction conditions and solvents used and probably also to the
variability associated with the difference in the stage of maturity of
cultivars and geographical area of origin [60]. According to Saad et al.
[61], total polyphenol levels increase at the early stage of growth in
peel, but thereafter generally decrease during maturation.
Yan et al. [57] found an ellagic acid content of 0.39–3.04 mg/g in
pomegranate peel extract and similarly, Seeram et al. [62] reported that
pomegranate peel contains major ellagitannins, such as punicalagin
(80–85% w/w) and ellagic acid (1.3% w/w). Zhou et al. [63] and Cam
and Hisil [1] identified ellagic acid and the other ellagic acid deriva-
tives, which were not determined in the present study. This may be
attributed to the smaller solubility of ellagic acid in aqueous ethanol
compared to methanol, which was used as an extraction solvent in the
aforementioned studies. Panichayupakarananta et al. [64] investigated
the influence of different methanol concentrations on the content of
ellagic acid in pomegranate peel extracts and concluded that 90% v/v
methanol gave significantly higher content of ellagic acid compared to
other analyzed methanol percentages, whereas Rodrigues et al. [65]
also showed that during UAE of jabuticaba tree, the best results for
ellagic acid were achieved at high ethanol concentration and high ex-
traction time. In addition, Elfalleh et al. [66] reported a gallic acid
content varied from 1.09 to 1.31 mg/g in six varieties of Tunisian po-
megranate peel after methanol extraction overnight. Generally, al-
though the existence of gallic acid, caffeic acid, chlorogenic acid, p-
coumaric acid, catechin, quercetin, and rutin in pomegranate peels
were reported in the literature, these polyphenols were not identified in
the present work. Gullon et al. [56] reported that the type and con-
centration of bioactive compounds in pomegranate peel extract depend
on several factors, such as cultivar analyzed, soil factors, environmental
conditions, maturity stages and so on. According to Bruni [67], the
proclivity to hydrolysis and in particular the different solubility in
water of some ellagitannins according to their molecular weight, are
factors that must be taken into account when comparing phytochemical
profiles of pomegranate peels in the literature. Generally, a number of
researchers have concluded that the solvent used, the extraction
method, and the duration of the process may severely affect both the
amount of phenolics and their relative ratios.
3.4. Comparison between microwave- and ultrasound-assisted extraction
Several researchers extracted phenolic compounds of pomegranate
peel using different extraction methods (Soxhlet, normal stirring,
pressurized water extraction, supercritical fluid extraction) [11] and
reported much longer extraction times (up to 60 times) to achieve
lower, similar or slightly higher extraction yields. Negi et al. [68] ex-
tracted phenolics from pomegranate peels using the Soxhlet method
with water for 4 h and reported a yield of 4.8%. Pan et al. [69] and
Wang et al. [70] used the normal stirring method with water and
achieved a total phenolic yield of 119 and 82.6 mg GAE/g dry matter at
8
Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11
60 and 240 min, respectively. Cam and Hisil [1] reported a maximum
yield of 264 mg tannic acid equivalents/g dry matter at 15 min with the
pressurized water extraction method. Mushtaq et al. [71] proposed an
enzyme-assisted supercritical fluid extraction of phenolics form pome-
granate peels with an optimum yield of 301.53 mg GAE/g extract at
85 min.
As far as new separation “green’’ techniques are concerned, ultra-
sound- and microwave-assisted extraction are energy-saving technolo-
gies that are increasingly employed in the extraction of natural products
as alternatives to traditional techniques of extraction. Pan et al. [13]
reported extraction yields, under the optimum conditions of ultrasound
extraction of phenolics from pomegranate peel with water, of 14.8 and
14.5% at extraction times of 6 and 8 min for continuous UAE and pulse
UAE, respectively. Tabaraki et al. [14] achieved an optimum yield of
91.98 mg GAE/g dry peels using 70% ethanol–water mixture as solvent
and extraction time of 30 min. In our previous work, Kaderides et al.
[11] studied the effects of various parameters on ultrasound-assisted
extraction yield of pomegranate peel phenolics and found the optimum
conditions to be: solvent type, water; extraction time, 10 min; tem-
perature, 34.7 °C; solvent/solid ratio, 32.2/1; amplitude level, 39.8%;
pulse duration/pulse interval ratio, 1.2/1. Under these optimized con-
ditions, the experimental value of extraction yield was 119.82 mg GAE/
g dry peels at an extraction time of 10 min. The obtained UAE extract
was analysed in the present work for polyphenol qualitative and
quantitative determination, antioxidant activity, and surface mor-
phology.
The microwave-assisted extraction method that was used in the
present work provided higher extraction efficiencies in comparison to
ultrasound-assisted extraction (Table 2). The MAE of pomegranate peel
phenolics had a maximum yield of 199.4 mg GAE/g dry peel at an ex-
traction time of 4 min A similar trend was observed by Ince et al. [72]
during the extraction of phenolic compounds from nettle by micro-
waves (24.64 mg GAE/g dry material) and ultrasounds (23.86 mg GAE/
g dry material). The solvent accessibility to the pomegranate peels must
have been enhanced due to the action of microwaves, disrupting the
peels matrix and allowing the penetration of solvent. Generally, ultra-
sound waves and the cavitation effect they produce can alter biological
materials, facilitating the release of extractable compounds and en-
hancing mass transport by disrupting cell walls, whereas the effec-
tiveness of microwaves is attributed to its localized heating which in-
creases the internal pressure of the cells and consequently ruptures
them. Heat and mass transfer occur in the same direction from inside to
outside in MAE, accelerating the solubilisation of solutes. Meanwhile
the heat that transferred from the outside to the inside of samples in the
ultrasounds method prolongs the extraction period needed for the so-
lutes to diffuse out and become solubilised in the solvent.
The raw material (RM) and the treated by MAE and UAE peels at the
optimum conditions were examined by scanning electron microscopy
(SEM) and their micrographs are shown in Fig. 7. The surface of the
peels sample before extraction was densely packed, whereas extraction
produced structural changes in all samples. After UAE, no significant
changes could be found for the surfaces of samples except for few
creases and ruptures, which may be due to the cavitation effects of
ultrasounds [73]. On the contrary, in the case of MAE, the sudden
thermal stresses and the high localized pressures affect the cell structure
of the cell causing intense creases and ruptures. Dahmoune et al. [74]
confirmed in their work on the microwave extraction of polyphenolic
compounds from medicinal plants that the electromagnetic waves
caused cell damage and rupture due to sudden temperature rise during
microwave irradiation. Alara et al. [37] also reported that microwave
irradiation plays an important role in vegetal cell wall breakage and
this phenomenon might be due to the rotation of liquids and the mo-
lecular movement associated with a permanent dipole, resulting in
rapid heating of the solvent. This structural change allows easy entry of
the solvent into cellular channels with high extraction efficiency.
In addition, the lower total phenolics content obtained by the
K. Kaderides et al. Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11

ultrasound-assisted method could be attributed to a possible degrada-

tion of the extract due to the generation of hydroxyl radicals by acoustic
Antioxidant activity (radical scavenging cavitation during ultrasounds treatment in water [75]. Vinatoru et al.
[76] reported that the presence of water vapor in the bubbles leads to a
homolytic splitting of the water molecules generating reactive HO and
hydrogen atoms during its collapse. The radicals formed then undergo
reactions to produce H2O2 and other active oxidising agents resulting in
possible phenolics degradation.
Punicalagin was found to be the major polyphenol in the optimized
pomegranate peel extract for both MAE and UAE, with a content of
143.64 and 138.8 mg/g dry matter, respectively (Fig. 6). Various po-
activity, %)

tential biological and pharmacological activities, including anti-bac-

terial, anti-mutagenic, anti-carcinogenic, anti-inflammatory, hepato-
protective, and anti-genotoxic properties, have been attributed to the
presence of punicalagin, mostly due to its high antioxidant activity
Punicalagin content (mg/g dry matter)

[56]. Gil et al. [77] determined the antioxidant activity of individual

94.77

phenolic groups in pomegranate juice by the DPPH method and found

that punicalagin had about 1.5-, 6.5-, and 20-times higher antioxidant
activity than those of hydrolyzable tannins, anthocyanins, and ellagic
94.91

acids, respectively, due to the presence of 16 phenolic hydroxyls per

molecule in its structure. According to Gil et al. [77], among the po-
Extract properties

megranate ellagitannins, punicalagin, which is the largest polyphenol,

having a molecular weight of greater than 1000, is reported to be re-
sponsible for more than half the potent antioxidant activity of the juice.
143.64

138.8

Moreover, Scalbert et al. [78] reported that phenolic compounds such

as caffeic acid can easily be absorbed through the gut barrier, however
large molecular weight compounds like punicalagin and proanthocya-
Extraction yield (mg GAE/g dry

nidins are very poorly absorbed. These findings are the most convincing
evidence that the antioxidant activity of pomegranate peel is mainly
related to the high contents of punicalagin in the extract. However,
there is a need for in depth study of quantitative aspect, due to possible
toxicity of this compound in large amounts of consumption.
Recent studies showed the antioxidant properties of different po-
Comparison between microwave- and ultrasound-assisted extraction of phenolics from pomegranate peels.

megranate peel extracts. Singh et al. [79] reported that a methanol

119.82
199.4
peel)

extract of pomegranate peels showed 81% antioxidant activity at

50 ppm using the DPPH model system. In this work, the radical
Temperature (oC)

scavenging activity of the optimized pomegranate peel extracts for MAE

and UAE was 94.91 and 94.77%, respectively. The slightly lower ac-
tivity of the UAE extract may be associated with the lower punicalagin
content and the solvent used (water instead of aqueous ethanol). Negi
35

et al. [68], who studied the antioxidant and antimutagenic activities of

various pomegranate peel extracts, mentioned that all the extracts
Solvent/peels (mL/g)

showed an increase in antioxidant capacity with an increase in phe-

nolics concentration and the water extract showed less antioxidant
capacity at all the concentrations.
Solvent/peels (mL/g)

32.2/1

4. Conclusions

Increased concern over the safety of synthetic antioxidants has led

to an increased interest in exploration of effective and economical
60/1

natural antioxidants. Pomegranate peel could be a good commercial

Amplitude (W)

source of phenolic compounds that can be separated and concentrated

Power (W)

through different extraction processes. The extract can be used as a

substitute of synthetic antioxidants for food products. In this work, a
600

method for pomegranate peels application in food industries based on

52

the microwave-assisted extraction of phenolics compounds was opti-

Time (min)

Time (min)

mized. Only 4 min in aqueous ethanol (a green environmental solvent)

Ultrasound-assisted extraction
Microwave-assisted extraction
Optimum extraction conditions

are needed to recover polyphenols from pomegranate peel with a high

10

yield (199.4 mg GAE/g dry peel), whereas conventional extraction

50% aqueous ethanol 4

procedures need much longer extraction times (up to 60 times). In

addition, microwaves lead to a higher extraction yield obtained in a
shorter process time (4 min) in comparison to another “green” extrac-
tion approach, the ultrasound-assisted method (119.82 mg GAE/g dry
Solvent

Solvent

peel in 10 min). This difference is attributed to the intense cell de-

Table 2

Water

struction of plant material observed by SEM analysis. The obtained

extract is characterized by a great antioxidant activity (radical

9
K. Kaderides et al.
Fig. 7. SEM images (×2000) of raw pomegranate peels before extraction (RM) and peels after microwave-assisted (MAE) and ultrasound-assisted (UAE) extraction at
the optimum conditions.
scavenging activity of 94.91%) due to the high content of punicalagin
(143.64 mg/g dry matter) measured by HPLC analysis. Thus, the pro-
posed method meets the terms of green process definition, since reduces
process time, allows use of alternative solvents (aqueous ethanol) and
renewable natural products, and ensure a safe and high quality extract/
product.
Acknowledgments
This research was in part supported by the General Secretariat for
Research and Technology (GSRT) and the Hellenic Foundation for
Research and Innovation (HFRI).
References
[1] M. Çam, Y. Hişil, Pressurised water extraction of polyphenols from pomegranate
peels, Food Chem. 123 (2010) 878–885.
[2] S.R. Kanatt, R. Chander, A. Sharma, Antioxidant and antimicrobial activity of po-
megranate peel extract improves the shelf life of chicken products, Int. J. Food Sci.
Technol. 45 (2010) 216–222.
[3] B.M. Naveena, A.R. Sen, S. Vaithiyanathan, Y. Babji, N. Kondaiah, Comparative
efficacy of pomegranate juice, pomegranate rind powder extract and BHT as anti-
oxidants in cooked chicken patties, Meat Sci. 80 (2008) 1304–1308.
[4] S. Iqbal, S. Haleem, M. Akhtar, M. Zia-ul-Haq, J. Akbar, Efficiency of pomegranate
peel extracts in stabilization of sunflower oil under accelerated conditions, Food
Res. Int. 41 (2008) 194–200.
[5] O.K. Topuz, P. Yerlikaya, I. Ucak, B. Gumus, H.A. Büyükbenli, Effects of olive oil
and olive oil-pomegranate juice sauces on chemical, oxidative and sensorial quality
of marinated anchovy, Food Chem. 154 (2014) 63–70.
[6] K.V. Kumudavally, A. Tabassum, K. Radhakrishna, A.S. Bawa, Indian Pat. Appl
2009 (2009) IN 2007DE01607 A 20090306.
[7] S.K. Devatkal, P.R. Thorat, M. Manjunatha, R.K. Anurag, Comparative antioxidant
effect of aqueous extracts of curry leaves, fenugreek leaves and butylated hydro-
xytoluene in raw chicken patties, J. Food Sci. Technol. 49 (2012) 781–785.
[8] M. Çam, F. Erdoǧan, D. Aslan, M. Dinç, Enrichment of functional properties of ice
cream with pomegranate by-products, J. Food Sci. 78 (2013) 1543–1550.
[9] J. Ventura, F. Alarcón-Aguilar, R. Roman-Ramos, E. Campos-Sepulveda, M.L. Reyes-
10
Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11
Vega, V.D. Boone-Villa, E.I. Jasso-Villagómez, C.N. Aguilar, Quality and antioxidant
properties of a reduced-sugar pomegranate juice jelly with an aqueous extract of
pomegranate peels, Food Chem. 136 (2013) 109–115.
[10] H. Wasila, X. Li, L. Liu, I. Ahmad, S. Ahmad, Peel effects on phenolic composition,
antioxidant activity, and making of pomegranate juice and wine, J. Food Sci. 78
(2013) 1166–1172.
[11] K. Kaderides, A.M. Goula, K.G. Adamopoulos, A process for turning pomegranate
peels into a valuable food ingredient using ultrasound-assisted extraction and en-
capsulation, Innov. Food Sci. Emerg. Technol. 31 (2015) 204–215.
[12] H.C. Tiwari, P. Singh, P.K. Mishra, P. Srivastava, Evaluation of various techniques
for extraction of natural colorants from pomegranate rind-ultrasound and enzyme
assisted extraction, Indian J. Fibre Textile Res. 35 (2010) 272–276.
[13] Z. Pan, W. Qu, H. Ma, G.G. Atungulu, T.H. McHugh, Continuous and pulsed ul-
trasound-assisted extractions of antioxidants from pomegranate peel, Ultrason.
Sonochem. 19 (2012) 365–372.
[14] R. Tabaraki, E. Heidarizadi, A. Benvidi, Optimization of ultrasonic-assisted ex-
traction of pomegranate (Punica granatum L.) peel antioxidants by response surface
methodology, Sep. Purif. Technol. 98 (2012) 16–23.
[15] R. Boggia, F. Turrini, C. Villa, C. Lacapra, P. Zunin, B. Parodi, Green extraction from
pomegranate marcs for the production of functional foods and cosmetics,
Pharmaceuticals 9 (2016) 1–11.
[16] X. Zheng, B. Liu, L. Li, Z. Zhu, Microwave-assisted extraction and antioxidant ac-
tivity of total phenolic compounds from pomegranate peel, J. Med. Plants Res. 5
(2011) 1004–1011.
[17] J. Huang, W. He, C. Yan, X. Du, X. Shi, Microwave assisted extraction of flavonoids
from pomegranate peel and its antioxidant activity, BIO Web of Conferences 8
(2017) 1–6.
[18] P.C. Veggi, J. Martinez, M.A.A. Meireles, Fundamentals of microwave extraction, in:
F.C, G. Cravotto (Eds.), Microwave-Assisted Extraction for Bioactive Compounds:
Theory and Practice, 1st ed., Springer US, New York, 2013, pp. 15–52.
[19] X.Z. Zheng, X.W. Xu, C.H. Liu, Y. Sun, Z. Lin, H.J. Liu, Extraction characteristics and
optimal parameters of anthocyanin from blueberry powder under microwave- as-
sisted extraction conditions, Sep. Purif. Technol. 104 (2013) 17–25.
[20] K. Hayat, S. Hussain, S. Abbas, U. Farooq, B. Ding, S. Xia, C. Jia, X. Zhang, W. Xia,
Optimized microwave-assisted extraction of phenolic acids from citrus mandarin
peels and evaluation of antioxidant activity in vitro, Sep. Purif. Technol. 70 (2009)
63–70.
[21] T.S. Ballard, P. Mallikarjunan, K. Zhou, S. O’Keefe, Microwave-assisted extraction of
phenolic antioxidant compounds from peanut skins, Food Chem. 120 (2010)
1185–1192.
[22] Y. Li, G.K. Skouroumounis, G.M. Elsey, D.K. Taylor, Microwave-assistance provides
very rapid and efficient extraction of grape seed polyphenols, Food Chem. 129
K. Kaderides et al.
(2011) 570–576.
[23] H. Li, Z. Deng, T. Wu, R. Liu, S. Loewen, R. Tsao, Microwave-assisted extraction of
phenolics with maximal antioxidant activities in tomatoes, Food Chem. 130 (2012)
928–936.
[24] M.D. Pavlović, A.V. Buntić, S.S. Šiler-Marinković, S.I. Dimitrijević-Branković,
Ethanol influenced fast microwave-assisted extraction for natural antioxidants ob-
taining from spent filter coffee, Sep. Purif. Technol. 118 (2013) 503–510.
[25] V.M. Simić, K.M. Rajković, S.S. Stojičević, D.T. Veličković, N.C. Nikolić, M.L. Lazić,
I.T. Karabegović, Optimization of microwave-assisted extraction of total poly-
phenolic compounds from chokeberries by response surface methodology and ar-
tificial neural network, Sep. Purif. Technol. 160 (2016) 89–97.
[26] A.R. Vieira, L. Abar, D.S.M. Chan, S. Vingeliene, E. Polemiti, C. Stevens,
D. Greenwood, T. Norat, Foods and beverages and colorectal cancer risk: a sys-
tematic review and meta-analysis of cohort studies, an update of the evidence of the
WCRF-AICR Continuous Update Project, Ann. Oncol. 28 (2017) 1788–1802.
[27] J. Pinela, M.A. Prieto, M.F. Barreiro, A.M. Carvalho, M.B.P.P. Oliveira,
J.A. Vázquez, I.C.F.R. Ferreira, Optimization of microwave-assisted extraction of
hydrophilic and lipophilic antioxidants from a surplus tomato crop by response
surface methodology, Food Bioprod. Process. 98 (2016) 283–298.
[28] S.R. Shirsath, S.S. Sable, S.G. Gaikwad, S.H. Sonawane, D.R. Saini, P.R. Gogate,
Intensification of extraction of curcumin from Curcuma amada using ultrasound
assisted approach: effect of different operating parameters, Ultrason. Sonochem. 38
(2017) 437–445.
[29] D.O. Kim, O.K. Chun, Y.J. Kim, H.Y. Moon, C.Y. Lee, Quantification of poly-
phenolics and their antioxidant capacity in fresh plums, J. Agric. Food Chem. 51
(2003) 6509–6515.
[30] C.Z. Shen, H.Y. Jun, S.H. Choi, Y.M. Kim, E.J. Jung, G.S. Oh, S.J. Joo, S.H. Kim,
I.K. Kim, Evaluation of antioxidant activities and active compounds separated from
water soluble extracts of Korean Black pine barks, Bull. Korean Chem. Soc. 31
(2010) 3567–3572.
[31] K. Kaderides, A.M. Goula, Extract from pomegranate waste as an alternative natural
antioxidant in foods. Bioprod. Process., 2019. Submitted for publication.
[32] M.M. Poojary, P. Passamonti, Extraction of lycopene from tomato processing waste:
kinetics and modelling, Food Chem. 173 (2015) 943–950.
[33] A.M. Goula, Ultrasound-assisted extraction of pomegranate seed oil - kinetic mod-
eling, J. Food Eng. 117 (2013) 492–498.
[34] J.P. Maran, V. Sivakumar, K. Thirugnanasambandham, R. Sridhar, Optimization of
microwave assisted extraction of pectin from orange peel, Carbohydr. Polym. 97
(2013) 703–709.
[35] J.H. Xie, C.J. Dong, S.P. Nie, F. Li, Z.J. Wang, M.Y. Shen, M.Y. Xie, Extraction,
chemical composition and antioxidant activity of flavonoids from Cyclocarya pa-
liurus (Batal.) Iljinskaja leaves, Food Chem. 186 (2015) 97–105.
[36] T. Wu, J. Yan, R. Liu, M.F. Marcone, H.A. Aisa, R. Tsao, Optimization of microwave-
assisted extraction of phenolics from potato and its downstream waste using or-
thogonal array design, Food Chem. 133 (2012) 1292–1298.
[37] O.R. Alara, N.H. Abdurahman, O.A. Olalere, Optimization of microwave-assisted
extraction of flavonoids and antioxidants from Vernonia amygdalina leaf using re-
sponse surface methodology, Food Bioprod. Process. 107 (2018) 36–48.
[38] C. Reichardt, Solvatochromic dyes as solvent polarity indicators, Chem. Rev. 94
(1994) 2319–2358.
[39] J.S. Boeing, É.O. Barizão, B.C. e Silva, P.F. Montanher, V. de Cinque Almeida,
J.V. Visentainer, Evaluation of solvent effect on the extraction of phenolic com-
pounds and antioxidant capacities from the berries: application of principal com-
ponent analysis, Chem. Cent. J. 8 (2014) 1–9.
[40] A.M. Galan, I. Calinescu, A. Trifan, C. Winkworth-Smith, M. Calvo-Carrascal,
C. Dodds, E. Binner, New insights into the role of selective and volumetric heating
during microwave extraction: investigation of the extraction of polyphenolic com-
pounds from sea buckthorn leaves using microwave-assisted extraction and con-
ventional solvent extraction, Chem. Eng. Process. Process. Intensif. 116 (2017)
29–39.
[41] D.C. Campos, E.L. Dall’Oglio, P.T. De Sousa, L.G. Vasconcelos, C.A. Kuhnen,
Investigation of dielectric properties of the reaction mixture during the acid-cata-
lyzed transesterification of Brazil nut oil for biodiesel production, Fuel 117 (2014)
957–965.
[42] R. Nagahata, T. Nakamura, Microwave-assisted rapid synthesis of poly(butylene
succinate): principal effect of microwave irradiation of accelerating the poly-
condensation reaction, Polym. J. 50 (2018) 347–354.
[43] F. Anwar, R. Przybylski, Effect of solvents extraction on total phenolics and anti-
oxidant activity of extracts from flaxseed (Linum usitatissimum L.), Acta Sci. Pol.
Technol. Aliment. 11 (2012) 293–302.
[44] G. Spigno, L. Tramelli, D.M. De Faveri, Effects of extraction time, temperature and
solvent on concentration and antioxidant activity of grape marc phenolics, J. Food
Eng. 81 (2007) 200–208.
[45] F.S. Taha, G.F. Mohamed, S.H. Mohamed, S.S. Mohamed, M.M. Kamil, Optimization
of the extraction of total phenolic compounds from sunflower meal and evaluation
of the bioactivities of chosen extracts, Am. J. Food Technol. 6 (2011) 1002–1020.
[46] M. Gfrerer, E. Lankmayr, Screening, optimization and validation of microwave-
assisted extraction for the determination of persistent organochlorine pesticides,
Anal. Chim. Acta 533 (2005) 203–211.
[47] J.P. Maran, V. Sivakumar, K. Thirugnanasambandham, R. Sridhar, Microwave as-
sisted extraction of pectin from waste Citrullus lanatus fruit rinds, Carbohydr.
Polym. 101 (2014) 786–791.
[48] S.Q. Liew, G.C. Ngoh, R. Yusoff, W.H. Teoh, Sequential ultrasound-microwave as-
sisted acid extraction (UMAE) of pectin from pomelo peels, Int. J. Biol. Macromol.
93 (2016) 426–435.
[49] J.P. Maran, K. Swathi, P. Jeevitha, J. Jayalakshmi, G. Ashvini, Microwave-assisted
extraction of pectic polysaccharide from waste mango peel, Carbohydr. Polym. 123
(2015) 67–71.
11
Chemical Engineering & Processing: Process Intensification 137 (2019) 1–11
[50] M. Ghafoor, N.N. Misra, K. Mahadevan, B.K. Tiwari, Ultrasound assisted hydration
of navy beans (Phaseolus vulgaris), Ultrason. Sonochem. 21 (2014) 409–414.
[51] J.B. Ji, X.H. Lu, M.Q. Cai, Z.C. Xu, Improvement of leaching process of Geniposide
with ultrasound, Ultrason. Sonochem. 13 (2006) 455–462.
[52] D.M. Patil, K.G. Akamanchi, Microwave assisted process intensification and kinetic
modelling: extraction of camptothecin from Nothapodytes nimmoniana plant, Ind.
Crops Prod. 98 (2017) 60–67.
[53] R. Yedhu Krishnan, K.S. Rajan, Microwave assisted extraction of flavonoids from
Terminalia bellerica: study of kinetics and thermodynamics, Sep. Purif. Technol. 157
(2016) 169–178.
[54] W. Qu, Z. Pan, H. Ma, Extraction modeling and activities of antioxidants from po-
megranate marc, J. Food Eng. 99 (2010) 16–23.
[55] F. Aqil, R. Munagala, M.V. Vadhanam, H. Kausar, J. Jeyabalan, D.J. Schultz,
R.C. Gupta, Anti-proliferative activity and protection against oxidative DNA da-
mage by punicalagin isolated from pomegranate husk, Food Res. Int. 49 (2012)
345–353.
[56] B. Gullon, M.E. Pintado, J.A. Pérez-Álvarez, M. Viuda-Martos, Assessment of
polyphenolic profile and antibacterial activity of pomegranate peel (Punica gran-
atum) flour obtained from co-product of juice extraction, Food Control 59 (2016)
94–98.
[57] L. Yan, X. Zhou, L. Shi, D. Shalimu, C. Ma, Y. Liu, Phenolic profiles and antioxidant
activities of six Chinese pomegranate (Punica granatum L.) cultivars, Int. J. Food
Prop. 20 (2017) 94–107.
[58] T. Ismail, P. Sestili, S. Akhtar, Pomegranate peel and fruit extracts: a review of
potential anti-inflammatory and anti-infective effects, J. Ethnopharmacol. 143
(2012) 397–405.
[59] J. Lu, K. Ding, Q. Yuan, Determination of punicalagin isomers in pomegranate husk,
Chromatographia 68 (2008) 303–306.
[60] S. Kharchoufi, F. Licciardello, L. Siracusa, G. Muratore, M. Hamdi, C. Restuccia,
Antimicrobial and antioxidant features of ‘Gabsiʼ pomegranate peel extracts, Ind.
Crops Prod. 111 (2018) 345–352.
[61] H. Saad, F. Charrier-El Bouhtoury, A. Pizzi, K. Rode, B. Charrier, N. Ayed,
Characterization of pomegranate peels tannin extractives, Ind. Crops Prod. 40
(2012) 239–246.
[62] N.P. Seeram, L.S. Adams, S.M. Henning, Y. Niu, Y. Zhang, M.G. Nair, D. Heber, In
vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic
acid and a total pomegranate tannin extract are enhanced in combination with
other polyphenols as found in pomegranate juice, J. Nutr. Biochem. 16 (2005)
360–367.
[63] B.H. Zhou, Z.H. Wu, X.J. Li, J. Zhang, X.M. Hu, Analysis of ellagic acid in pome-
granate rinds by capillary electrophoresis and high-performance liquid chromato-
graphy, Phytochem. Anal. 19 (2008) 86–89.
[64] P. Panichayupakarananta, A. Issuriya, A. Sirikatitham, W. Wang, Antioxidant assay-
guided purification and LC determination of ellagic acid in pomegranate peel, J.
Chromatogr. Sci. 48 (2010) 456–459.
[65] S. Rodrigues, F.A.N. Fernandes, E.S. de Brito, A.D. Sousa, N. Narain, Ultrasound
extraction of phenolics and anthocyanins from jabuticaba peel, Ind. Crops Prod. 69
(2015) 400–407.
[66] W. Elfalleh, H. Hannachi, N. Tlili, Y. Yahia, N. Nasri, A. Ferchichi, Total phenolic
contents and antioxidant activities of pomegranate peel, seed, leaf and flower, J.
Med. Plants Res. 6 (2012) 4724–4730.
[67] R. Bruni, Pomegranate phytochemicals: distribution and variability, in: A. Caligiani
(Ed.), Pomegranate Chemistry, Proceeding and Health Benefits, 1st ed., Nova
Science Publishers, USA, 2016, pp. 27–56.
[68] P.S. Negi, G.K. Jayaprakasha, B.S. Jena, Antioxidant and antimutagenic activities of
pomegranate peel extracts, Food Chem. 80 (2003) 393–397.
[69] Z. Pan, W. Qu, H. Ma, G.G. Atungulu, T.H. McHugh, Continuous and pulsed ul-
trasound-assisted extractions of antioxidants from pomegranate peel, Ultrason.
Sonochem. 18 (2011) 1249–1257.
[70] Z. Wang, Z. Pan, H. Ma, G.G. Atungulu, Extract of phenolics from pomegranate
peels, Open Food Sci. J. 5 (2011) 17–25.
[71] M. Mushtaq, B. Sultana, F. Anwar, A. Adnan, S.S.H. Rizvi, Enzyme-assisted super-
critical fluid extraction of phenolic antioxidants from pomegranate peel, J.
Supercrit. Fluids 104 (2015) 122–131.
[72] A.E. Ince, S. Sahin, G. Sumnu, Comparison of microwave and ultrasound assisted
extraction techniques for leaching of phenolic compounds from nettle, J. Food Sci.
Technol. 51 (2014) 2776–2782.
[73] M. Romdhane, C. Gourdon, Investigation in solid-liquid extraction: influence of
ultrasound, Chem. Eng. J. 87 (2002) 11–19.
[74] F. Dahmoune, B. Nayak, K. Moussi, H. Remini, K. Madani, Optimization of micro-
wave-assisted extraction of polyphenols from Myrtus communis L. Leaves, Food
Chem. 166 (2015) 585–595.
[75] Z. Lianfu, L. Zelong, Optimization and comparison of ultrasound/microwave as-
sisted extraction (UMAE) and ultrasonic assisted extraction (UAE) of lycopene from
tomatoes, Ultrason. Sonochem. 15 (2008) 731–737.
[76] M. Vinatoru, T.J. Mason, I. Calinescu, Ultrasonically assisted extraction (UAE) and
microwave assisted extraction (MAE) of functional compounds from plant mate-
rials, Trends Analyt. Chem. 97 (2017) 159–178.
[77] M.I. Gil, F.A. Tomas-Barberan, B. Hess-Pierce, D.M. Holcroft, A.A. Kader,
Antioxidant activity of pomegranate juice and its relationship with phenolic com-
position and processing, J. Agric. Food Chem. 48 (2000) 4581–4589.
[78] A. Scalbert, C. Morand, C. Manach, C. Rémésy, Absorption and metabolism of
polyphenols in the gut and impact on health, Biomed. Pharmacother. 56 (2002)
276–282.
[79] R.P. Singh, K.N.C. Murthy, G.K. Jayaprakasha, Studies on the antioxidant activity of
pomegranate (Punica granatum) peel and seed extracts using in vitro models, J.
Agric. Food Chem. 50 (2002) 81–86.