1386 International Journal of Food Science and Technology 2010, 45, 1386–1394

Original article
Encapsulation of polyphenols and anthocyanins from pomegranate
(Punica granatum) by spray drying

Paz Robert,1* Tamara Gorena,2 Nalda Romero,1 Elena Sepulveda,2 Jorge Chavez3 & Carmen Saenz2
1 Depto. Ciencia de los Alimentos y Tecnologı́a Quı́mica, Facultad de Ciencias Quı́micas y Farmacéuticas, Universidad de Chile, Casilla 133,
Santiago, Chile
2 Depto. Agroindustria y Enologı́a, Facultad de Ciencias Agronómicas, Universidad de Chile, Casilla 1004, Santiago, Chile
3 Depto. Tecnologı́a Farmacéutica. Facultad de Ciencias Quı́micas y Farmacéuticas, Universidad de Chile, Casilla 133, Santiago, Chile
(Received 9 January 2010; Accepted in revised form 18 March 2010)

Summary Pomegranate (Punica granatum) bioactive compounds (polyphenols and anthocyanins) of juice (PJ) and
ethanolic extracts (PE) were encapsulated with maltodextrin (MD) or soybean protein isolates (SPI) by spray
drying using a 22 statistical factorial design for each systems studied (PJ–MD, PJ–SPI, PE–MD and PE–SPI)
considering the proportion of coating material and the inlet temperature as independent variables. The
stability of the bioactive compounds microcapsules powders obtained under optimal conditions for each
system was studied at 60 C in oven for 56 days. The polyphenols encapsulating efficiency was significantly
better in SPI matrix whereas for anthocyanins was in MD matrix. By the other hand, during the storage, the
MD microcapsules provided a significant greater protective effect on the polyphenols and anthocyanins than
SPI, as was shown by the lower degradation rate constants. When the microcapsules were added to yogurt
the stability of the bioactive compounds followed a similar behaviour to those without encapsulation, except
for PE–MD.
Keywords Anthocyanin, antioxidant activity, microcapsules, phenolic compounds, pomegranate.

Newman, 2007). Flavonols and anthocyanins show anti-

Introduction
carcinogenic, antimicrobial (Opara et al., 2009), anti-
Pomegranate originated from the Middle East, extend- inflammatory and antioxidant activities (Lansky &
ing throughout the Mediterranean area, eastward to Newman, 2007). The phenyl propanoids as chlorogenic,
China and India, and on to the American Southwest, caffeic and coumaric acids may be responsible of the
California and México. The pomegranate tree thrives inhibition of tumour initiation and development in rats
under arid and semiarid climatic conditions. The fruit is (Huang et al., 2005).
often deemed to be a large berry, generally harvested The pomegranate juice (PJ) is a source of antocyani-
when fully ripe, and it possesses a waxy shiny surface of dins such as 3-glucosides and 3,5 diglucosides of
reddish yellow according to the varieties (Mars et al., delphinidin, cyanidin and pelargodinin. The phenolic
1997). The fruit contain a large number of arils (seeds compounds as gallagyl-type tannins (punicalagin); ella-
surrounded by a translucent juice sac) from which a gic acid derivatives (ellagic acid glucoside and ellagic
valuable juice is obtained (Gil et al., 1995a; Magerranov acid); hydrolysable tannins (galloyl glucose and other
et al., 2007). Pomegranate has been used for centuries in compounds) have been reported (Nawwar et al., 1994;
ancient cultures in folk medicine. All parts of the Gil et al., 2000). Catechins, ellagitannins, gallitannins
pomegranate plant (bark, stem, arils, whole fruit juice and quercetin glycosides have been reported in PJ too
and leaves) have showed antioxidant activity. Studies (Noda et al., 2002). It also contains organic acids such as
about pomegranate polyphenols have shown prevention citric, ascorbic, malic and oxalic and small amounts of
of cardiovascular disease, cancer and neurological pectin. Different punicalagin content between commer-
damage in humans (Aviram et al., 2002; Mertens- cial and experimental juices (only arils) have been
Talcott et al., 2006; Seeram et al., 2006; Lansky & reported, because the industrial process includes pome-
granate rinds. Commercial PJ has one of the highest
*Correspondent: Fax: +56 02 2227900; antioxidant activities compared to other fruit juices, red
e-mail: proberts@uchile.cl wine and green tea (Gil et al., 2000).

doi:10.1111/j.1365-2621.2010.02270.x
 2010 Institute of Food Science and Technology
 Encapsulation of polyphenols and anthocyanins P. Robert et al. 1387

Pomegranate is an interesting source of polyphenols

Juice and ethanolic extract preparation
and anthocyanins which can be used as bioactive
ingredients in foods. However, these molecules are Juice preparation
unstable and the fresh juice has a short shelf life. In this The arils from the pomegranate were manually ob-
context, the stabilisation of polyphenol and anthocyanin tained. The juice (4.07 L) from a 5.985 kg aril blend was
compounds for use in industrial purposes could be aided obtained using a juice extractor (Moulinex T140-02).
using microencapsulation technologies (Desai & Park,
2005). Microencapsulation is described as a technique Ethanolic extract preparation
wherein a bioactive compound is encapsulated by a The aril blend (600.41 g) was macerated with ethanol
biopolymer thereby protecting it from oxygen, water, and water (1:1 w ⁄ w) (the addition of water included the
light or other conditions in order to improve its stability arils water content) during a total of 12 h. Three
and also to change liquid solutions to powders for easier extractions were made until the pulp was light red.
handling (Gharsallaoui et al., 2007). Encapsulation of Extracts were combined and concentrated in a Buchi
quercetin has also been reported for increase its water RE120 evaporator at 40 C until reaching a similar
solubility (Wu et al., 2008). polyphenol content of the juice, obtaining 830 mL of
The encapsulating agents used in this study were PE. The juice and extract was frozen at )20 C.
maltodextrin (MD) and soybean protein isolate (SPI).
Maltodextrins of different dextrose equivalent (DE) are
Preparation of the microcapsules
commonly used as wall material by its high water,
solubility, low viscosity, low sugar content and their Encapsulation in MD or SPI were prepared as follows:
solutions are colourless. These properties make them PJ (20 g for MD or 6 g for SPI) or ethanol extract (24 g
useful ingredients in the food industry (Avaltroni et al., for MD or 7.5 g for SPI) was mixed with MD (10–40%)
2004). Soybean protein is one of the most popular or SPI (3–12%) previously heated at 40 C, with
plant protein sources used as an ingredient in food constant stirring. The SPI percentage used in the
formulation. The globulins glycinin and b-conglycinin formulation before spray drying was lower than MD
are the major components of soybean isolates. These due to its high viscosity. Each preparation was homo-
two globulins have different structures and functional genised with an Ultraturrax IKA T50 at 1400 · g for
properties (Arrese et al., 1991; Pretuccelli & Añón, 5 min. The resultant solutions were fed to a mini spray
1996). Soybean protein isolate has been used for their dryer B191 (Büchi, Flawil, Switzerland). The spray
encapsulating and emulsifier characteristics in orange dryer was operated at inlet temperature ranging from
oil microcapsules, showing higher oil retention than 140 to 160 ± 5 C for MD and 100 to 140 ± 5 C for
whey protein isolate and arabic gum (Kim & Morr, SPI. The air flow, rate of feeding and atomisation
1996). pressure was 600 L h)1, 10 mL min)1 and 20 psi,
The objective of this research was to encapsulate PJ respectively, for both encapsulating agents. The powders
and ethanolic extract (PE) with MD or SPI by spray obtained were kept at )20 C until analysed.
drying, to study the effect of the inlet temperature and
the encapsulating agent content on the polyphenols and
Pomegranate juice and ethanolic extract analysis
anthocyanins encapsulation (retention), and to analyse
the matrix influence on the active compounds stability The soluble solids (Brix), pH, and acidity were deter-
of the obtained powders. mined according to AOAC methods (AOAC, 1996). The
total sugars were determined by the Antrona method
(Osborne & Voogt, 1986) in an UNICAM UV ⁄ VIS
Materials and methods
spectrometer UV3.
The total phenolic content was determined according
Materials
to the Folin Ciocalteau colorimetric method (Singleton
Pomegranate fruits (P. granatum) were obtained from a & Rossi, 1965), and the results were expressed as
plantation located in the ‘Las Cardas’ Experimental milligram of gallic acid equivalents (GAE), according
Station that belongs to the University of Chile, Ovalle, to a calibration curve (133.8–428.0 lg mL)1;
Chile. Two pomegranate fruit genotypes (PG2 and PG3) R2 = 0.9901). The total anthocyanins were determined
were selected by their high bioactive compound content spectrophotometrically at 520 nm (Giusti & Wrolstad,
and a blend of arils of PG2:PG3 at a ratio of 1:2.5 (w ⁄ w) 2001) and expressed as milligram of malvidin-3-gluco-
was used for juice and extract preparation. Maltodextrin side equivalents per L. The identification was performed
from corn (MD) (DE = 12–20) (Inducorn, Santiago, by HPLC using a Merck Hitachi L6200 pump, a Waters
Chile) and SPI (Prinal, Santiago, Chile) were used as 996 photodiode-array detector, and a C18 column
wall materials. All other reagents were of analytical (5 lm · 4.6 mm i.d. · 25 cm, YMCTM Carotenoid
grade. S-5, Waters, Milford, USA). The mobile phases used

 2010 Institute of Food Science and Technology International Journal of Food Science and Technology 2010, 45, 1386–1394
1388 Encapsulation of polyphenols and anthocyanins P. Robert et al.

were solvent A [formic acid:water (10:90 v ⁄ v)] and using EDS 7424 software (Oxford Instruments, Oxford,
solvent B (methanol) according to a program described UK).
by Prieto et al. (2005).
The antioxidant activity was evaluated in accordance
Accelerated storage stability test
with the radical scavenging DPPH method (Gil et al.,
2000). All the analyses were carried out in duplicate and Microcapsules of pomegranate (juice or ethanol extract)
averaged. obtained under optimal conditions for each studied
system (PJ–MD, PJ–SPI, PE–MD and PE–SPI) were
stored at 60 C in a forced-air oven (Memmert model
Microcapsule powder analysis
BE 500, Schwabach, Germany) with controlled temper-
Total bioactive compounds ature and in absence of light for 56 days. Samples of
The coating material structure of the microcapsule was 0.2 g of each powder were transferred to 100 · 150 mm
completely destructed by the following procedure: for clear glass vials. For determination of bioactive com-
MD, 200 mg of the microcapsules were accurately pounds (polyphenols and anthocyanins), duplicate vials
weighed and 2 mL of methanol:acetic acid:water were removed every 7 days. Pomegranate juice without
(50:8:42 v ⁄ v ⁄ v) was added. This dispersion was agitated encapsulation was used as a control.
using a Vortex (1 min) and then an ultrasonicator twice
for 20 min. The supernatant was centrifuged at
Addition of microcapsules to a yogurt
112 000 g for 5 min and then filtered. For SPI, 200 mg
of the microcapsules were accurately weighed, and 1 mL Glass jars containing 50 g of natural yogurt Next were
of acetonitrile and 1 mL of methanol:acetic acid:water added with 0.5 g of pomegranate encapsulated powders
(50:8:42 v ⁄ v ⁄ v) were added, and then the same proce- (obtained under optimal conditions) and storage at
dure described for MD was carried out. The amounts of 5 ± 1 C during 30 days in the absence of light. A
phenolic and anthocyanin compounds were quantified control was prepared with concentrated PJ (60 Brix).
as described above. Samples were removed once a week for the determina-
tion of bioactive compounds according to Coisson et al.
Surface bioactive compounds (2005). The yogurt assay was performed by triplicate.
For the determination of surface anthocyanin and
phenolic compounds, 200 mg of microcapsules were
Statistical design
treated with 2 mL of a mixture of ethanol and methanol
(1:1). These dispersions were agitated in a Vortex at The experiments were performed with a 22 central
room temperature for 1 min and then filtered (0.45 lm composed experimental design constituted by ten exper-
Millipore filter). The amounts of phenolic and anthocy- iments for each encapsulating agent. The independent
anin compounds were quantified as described above. The variables considered were the temperature of drying
surface bioactive compound percentage (polyphenols or (140–160 and 100–140 C for MD and SPI, respectively)
anthocyanins) and the microencapsulation efficiency and the coating material (10–40% and 3–12% for MD
(ME) of microencapsulated bioactive compounds were and SPI, respectively). The dependent variables were
calculated according to eqns 1 and 2, respectively. ME of microencapsulated anthocyanins and polyphe-
nols. Response surface methodology was applied to
surface bioactive compounds optimise the ME of the bioactive compounds using
SBð%Þ ¼  100 Statgraphics software version 7.0 (Manugistics Inc.,
theoretical total bioactive compounds
Statistical Graphics Corporation, Rockville, MA,
ð1Þ USA).

MEð%Þ ¼ 100  SBð%Þ ð2Þ Results and discussion

Juice and extract characteristics and bioactive compounds

Scanning electron microscopy Table 1 shows the physical and chemical analysis of the
The outer structures of the microcapsules obtained PJ and PE. The soluble solids and total sugar contents
under optimal conditions were studied by scanning of PJ from arils were 15.97 Brix and 17.72%, respec-
electron microscopy (SEM). The samples were coated tively, similar to those reported for thirteen varieties of
with gold ⁄ palladium using a Varian Vacuum Evapo- PJ from Turkey (16–19 Brix and 13.96–16.06%, respec-
rator PS 10E and analysed using a LEO 1420VP (LEO tively) (Poyrazoglu et al., 2002). The soluble solids were
Electron Microscopy Ltd, Cambridge, UK) operated at also similar to those reported by Alighourchi et al.
20 kV. The scanned images were collected digitally (2008) (12.1–18.33 Brix), higher than Magerranov et al.

International Journal of Food Science and Technology 2010, 45, 1386–1394  2010 Institute of Food Science and Technology
 Encapsulation of polyphenols and anthocyanins P. Robert et al. 1389

Table 1 Physical and chemical characteristics of PJ and PE from 2003) and lower than in a sweet juice from Spanish
pomegranate pomegranate cv. Mollar (2750 mg GAE L)1) (Pérez-
Vicente et al., 2004). Using other phenolic standard, Gil
PJ PE
et al. (2000) reported a phenolic content of 2117
Total soluble solids (Brix) 15.97 ± 0.06 13.80 ± 0.00 (mg L)1 p-cumaric acid) from a fresh the juice from
Total sugars (%) 17.72 ± 0.22 10.52 ± 0.31 arils cv. Wonderful of California and Yunfeng et al.
pH 3.15 ± 0.07 3.03 ± 0.01 (2006) reported a value of 24.4 mg tannic acid g)1 for
Acidity (% citric acid) 2.79 ± 0.13 1.02 ± 0.07 the phenolic content of pomegranate pulp.
Total phenolics compounds 2128.6 ± 0.01 1717.9 ± 5.05 The PJ total anthocyanin content (882.3 mg malvidin-
(mg GAE L)1)
3-glucoside L)1) was higher than that reported by
Total anthocyanins (mg malvidin 882.3 ± 0.01 552.1 ± 6.12
Tzulker et al. (2007), which was between 100 and
3-glucoside L)1)
Antioxidant activity 2.12 1.73
300 mg of malvidin-3-glucoside L)1 for juices prepared
DPPH (IC50 mg)1 mL)1) from the arils of twenty-nine Israeli pomegranate
accessions. Other authors have reported that the antho-
PJ, pomegranate juice; PE, pomegranate ethanolic extract; GAE, gallic cyanin content of pomegranates juices expressed as the
acid equivalent. sum of the individual anthocyanin contents, reached
values of 6–120 mg cyanidin-3-glucoside L)1 in juices
from Tunisian pomegranates (Gil et al., 1995b) and of
(2007) (11 Brix) and lower than Gil et al. (1995a) 162–387 mg cyanidin-3-glucoside L)1 in juices from
(18.25 Brix). The PE-soluble solids and total sugars fresh and frozen arils, and commercial juices from
contents were lower than those found for PJ due to a Californian pomegranates (Gil et al., 2000).
smaller extraction of those components during the The PE showed a lower total phenolic content
extract preparation. (1717.9 mg of EAG g)1) and total anthocyanins
Several organic acids such as citric, malic, oxalic, (552.1 mg of malvidin-3-glucoside L)1) compared with
acetic, fumaric, lactic and tartaric acids have been found the PJ, suggesting an incomplete extraction due to tissue
in pomegranate fruit, being citric acid the main. The PJ complexity and the different polarity and ⁄ or solubility
acidity was higher than to those informed by other of the polyphenols in ethanol.
authors, who reported to be 0.045–0.414% (Alighourchi The anthocyanins identified in JP and PE were
et al., 2008) and 0.46–1.73% (Poyrazoglu et al., 2002) delphinidin-3,5-diglucoside, cyanidin-3,5-diglucoside,
expressed as citric acid. The pH was reported to be 3.04– delphinidin-3-glucoside, pelargonidin-3,5-glucoside and
4.07 (Alighourchi et al., 2008) and 3.29–3.93 (Poyrazo- cyanidin-3-glucoside as can be seen in Fig. 1A,B. These
glu et al., 2002), similar to the value found in this study results showed the same anthocyanin profile as those
for PJ. reported by Miguel et al. (2004) for ‘Assaria’ pome-
The PJ total phenolic content reached values of 2128.6 granate, by Gil et al. (2000) for ‘Wonderful’ pomegran-
(mg GAE L)1), higher than the value reported in PJ cv. ate, and by Noda et al. (2002) who reported that
Suruc of Turkey (1564 mg L)1) (Vardin & Fenercioglu, delphinidin-3,5-diglucoside was a major anthocyanin

(A) (B)
ab a

b
Figure 1 Chromatograms of pomegranate ju-
ice (A) and ethanolic extract (B), determined cd c d
by HPLC. (a) Delphinidin-3,5-diglucoside; (b)
cyanidin-3,5-diglucoside+delphinidin-3-glu-
coside; (c) pelargonidin-3,5-glucoside and (d)
cyanidin-3-glucoside.

 2010 Institute of Food Science and Technology International Journal of Food Science and Technology 2010, 45, 1386–1394
1390 Encapsulation of polyphenols and anthocyanins P. Robert et al.

in juices from Californian pomegranates. In a recent 96.7–100%, respectively. In general, the encapsulation
paper, Alighourchi et al. (2008) in fifteen Iranian efficiency reached higher values for anthocyanin than
pomegranate varieties reported the presence of pelarg- polyphenol, showing the ability of MD and SPI to bind
onidin-3-glucoside too. A reduction in the cyanidin-3,5- anthocyanins. Thus, the flavylium cation could be
diglucoside+delphinidin-3-glucoside peak was observed related with the better polymer–anthocyanin interac-
in PE according with the lower anthocyanins content tion. By the other hand, Ersus & Yurdagel (2007)
observed in PE than PJ. studied the microencapsulation of anthocyanins pig-
The antioxidant activity (diphenylpicryl-hydrazyl ments of black carrot by spray drying using MD with
(DPPH)) of PJ and PE expressed as IC50 values were different DE and found that the greatest pigment
2.12 and 1.73 mg mL)1, respectively. Ricci et al. (2006) retention was when used 20–23 DE.
reported IC50 values for PJ (1.777 mg mL)1) and The response surface methodology was applied to
aqueous extract (2.511 mg mL)1, expressed as dry optimise the encapsulation efficiency of polyphenols and
weight). The greater PE antioxidant activity could be anthocyanins considering the linear, quadratic and
attributed to other compounds extracted simultaneously cross-product forms for the independent variables stud-
(i.e. tocopherols) with ethanol and that would be ied (encapsulating agent and temperature) at P £ 0.05
playing an antioxidant function too. levels, for each system. In the case of polyphenols
encapsulation, the encapsulating agent (A) linear form
was significant in all the systems studied and their
Pomegranate bioactive compounds microencapsulation
quadratic form (A2) had significant effect for PJ–SPI
The ME of polyphenols in PJ–SPI, PJ–MD, PE–SPI and PJ–MD. The temperature (B) linear form had a
and PE–MD microcapsules were in the range of 36.6– significant effect for PJ–MD and PE–SPI and their
62.8%, 51.4–82.8%, 52–81.5% and 52.9–82.8%, respec- quadratic form (B2) for PJ–MD. The cross-product
tively. Similar polyphenols encapsulation efficiency has form (AB) was only significant for PJ–SPI.
been reported by our group in previous studies about For anthocyanins encapsulation, the linear form (A)
microencapsulated cactus pear juice with MD (10 DE) was significant for PJ–SPI and the quadratic form (A2)
(39.41–74.78%) (Sáenz et al., 2009). Wu et al. (2008) for PJ–SPI, PE–SPI and PE–MD. The temperature
using other encapsulation method (nano-precipitation linear form (B) was only significant for PJ–SPI and their
technique), reported an efficiency over 94% in micro- quadratic form (B2) did not have effect as well as the
encapsulated quercetin using Eudragit and polyvinyl crossproduct form (AB). Therefore, the encapsulating
alcohol as carriers. Other researchers reported for load agent was the most important variable for the polyphe-
ME values of 48.5–87.1% in microencapsulated po- nols encapsulation whereas for the anthocyanin encap-
lyphenols from Illex paraguariensis, with calcium algi- sulation a smaller effect was observed according to its
nate and calcium alginate–chitosan (Deladino et al., high ME.
2008). Figure 2a–d shows the graphs obtained with the
The encapsulation efficiency of anthocyanins in PJ– response surface methodology for the PJ–SPI, PJ–MD,
SPI, PJ–MD, PE–SPI and PE–MD microcapsules were PE–SPI and PE–MD designs, respectively. The effect of
in the range of 35.8–100%, 89.4–100%, 73–98.9% and the temperature and the encapsulating agent on the

(a) (b)

(c) (d)

Figure 2 Graphs obtained by response sur-

face methodology for PJ–SPI (a), PJ–MD (b),
PE–SPI (c) and PE–MD (d).

International Journal of Food Science and Technology 2010, 45, 1386–1394  2010 Institute of Food Science and Technology
 Encapsulation of polyphenols and anthocyanins P. Robert et al. 1391

encapsulation efficiency (expressed as desirability) is allowing a greater bioactive compounds trapping than
shown. in PJ–SPI system. A higher core material to coating
material ratio (3:1) for encapsulated cactus pear juice and
ethanolic extract using MD (12–20 DE) was reported
Microcapsule powder obtained under optimal conditions
previously (Saénz et al., 2009). In that research, the
Table 2 contains the concentration values of polyphe- core ⁄ coating ⁄ water ratio and interaction between bioac-
nols and anthocyanins in the formulation and in the tive compounds and the coating material were discussed
spray-dried powder obtained under optimal conditions as parameters affecting encapsulation efficiency.
from the encapsulated juice and ethanolic extract. The The polyphenols encapsulating efficiency was signifi-
recovery percentages of bioactive compounds (polyphe- cantly better in SPI matrix whereas for anthocyanins
nols and anthocyanins) in all systems studied (PJ–SPI, was in MD matrix. This behaviour could be related with
PE–SPI, PJ–MD and PE–MD) were above 90%, the bioactive compounds nature (i.e. charge: negative
showing that the drying temperature did not affect the for polyphenols and positive for anthocyanins) and with
recovery of the bioactive compound. The recovery of polyelectrolyte structure (type and density charge),
polyphenols over 100% could be a consequence of the being SPI a poli(aminoacid) and MD a poli(glucosac-
hydrolysis of pomegranate polyphenols conjugated charide)), conditioning the bioactive–polymer interac-
during the preparation of the samples or during the tion. Kim & Morr (1996) determined that SPI shows the
drying process (Turkmen et al., 2005). In the b-carotene better encapsulating efficiency of orange oil, respect to
encapsulation with 25 DE MD, losses of 11% were whey protein isolate, arabic gum and sodium caseinate
obtained (Desobry et al., 1997). as encapsulating agents.
Table 3 shows the optimal conditions (percentage of Figure 3a–d presents SEM photographs of micro-
encapsulating agent and inlet temperature) and the ME capsules for the PJ–SPI, PJ–MD, PE–SPI and PE–MD
of the pomegranate bioactive compound microcapsules systems, respectively. The morphology of microcapsules
for PJ–SPI, PE–SPI, PJ–MD and PE–MD systems. The with both encapsulating agents was irregularly spherical
core material to coating material ratio was 1:1 for PJ– in shape with an extensively dented surface, attributed
SPI, PJ–MD and PE–MD systems and 2:1 for PE–SPI to the shrinkage of the particles during the drying
system. These results suggests that the interaction process. Similar morphology was observed in micro-
between the bioactive compounds and the coating capsules with different DE MDs (Dı́az et al., 2006;
material could be more important in the PE–SPI system Cai & Corke, 2000).

Table 2 Bioactive compounds before and after the encapsulation of the juice and ethanolic extract of pomegranate in SPI or MD

Polyphenols Anthocyanins

Theorical Theorical Experimental

System mg GAE g)1 Experimental % Recovery mg malvidin 3-glucoside g)1 % Recovery

PJ–SPI 1.53 1.85 ± 0.023 121 0.64 0.59 ± 0.003 92

PJ–MD 1.84 1.76 ± 0.033 96 0.76 0.74 ± 0.023 97
PE–SPI 2.75 2.84 ± 0.182 103 0.83 0.84 ± 0.022 101
PE–MD 1.53 1.51 ± 0.014 99 0.46 0.47 ± 0.007 102

PJ, pomegranate juice; PE, pomegranate ethanolic extract; SPI, soybean protein isolates; MD, maltodextrin; GAE, gallic acid equivalent.

Table 3 Optimal conditions and ME of the pomegranate bioactive compounds microcapsules using SPI or MD

Polyphenols Antocyanins
Core ⁄ coating encapsulated encapsulated
System Temperature (°C) EA (%) material (% ME) (% ME)

PJ–SPI 120 7.5 (1:1) 76.2b 58.5a

PJ–MD 153 20.1 (1:1) 53.5a 86.6c
PE–SPI 100 3.75 (1:1) 82.9c 100d
PE–MD 153 24.2 (2:1) 71.0b 82.0b

PJ, pomegranate juice; PE, pomegranate ethanolic extract; SPI, soybean protein isolates; MD, maltodextrin; EA, encapsulating agent; ME,
microencapsulation efficiency.
Different letters show significant differences between systems (by column) (P < 0.05).

 2010 Institute of Food Science and Technology International Journal of Food Science and Technology 2010, 45, 1386–1394
1392 Encapsulation of polyphenols and anthocyanins P. Robert et al.

(a) (b)

(c) (d)

Figure 3 Scanning electron microscopic

photographs of microcapsules for the PJ–SPI
(a), PJ–MD (b), PE–SPI (c) and PE–MD (d)
designs.

whey, protein isolate, arabic gum and sodium caseinate

Storage stability evaluation
as encapsulating agents. Dextrose equivalent of MDs
Figure 4 shows the evolution of polyphenols and has showed effect on the encapsulation yield but not
anthocyanins retention percentages during the storage effect was found during the storage in microencapsulat-
at 60 C for PJ–SPI, PE–SPI and PE–MD systems. ed black carrot anthocyanins (Ersus & Yurdagel, 2007).
When SPI is used as a wall material both polyphenol The polyphenols and anthocyanins degradation in fresh
and anthocyanin retention increase during the storage juice (Fig. 4) was faster than that in microencapsulated
time (until 35 days), possibly due to the hydrolysis of juice, showing the importance of the encapsulating
the pomegranate conjugated polyphenols, and then material in the degradation of bioactive compounds as
decrease. Similar behaviour was also reported by other has been reported for other functional compounds
authors (Stewart et al., 2000; Turkmen et al., 2005). The (Wagner & Warthesen, 1995).
polyphenol and anthocyanin retention diminish with The PJ system showed a significant lower degradation
storage time in MD microcapsules, except for PJ–MD of polyphenols and anthocyanins than the PE system,
system, where the polyphenol remains constant. Table 4 suggesting that some components of the juice could aid
shows the first-order degradation rate constant for the encapsulating process and therefore the stability of
polyphenols and anthocyanin encapsulated, calculated the bioactive compounds.
between 35 and 56 days. PJ–MD system showed the Table 4 shows the polyphenols degradation rate
lowest degradation rate for polyphenols and anthocya- constant in yogurt for PJ–MD, PJ–SPI, PE–MD and
nins encapsulated. PJ–SPI, PE–MD and PE–SPI PE–SPI systems. No statistical differences were observed
systems had higher polyphenols degradation rate how- between control (7.58 ± 0.73 day)1) and PJ–MD, PJ–
ever there was not significant differences between them. SPI and PE–SPI. The encapsulating agent (MD or SPI)
The anthocyanins show significant differences between showed significant effect on the polyphenols degradation
all systems, being the highest degradation rate for the rate constant for PE systems, having the MD the highest
PE–SPI system, followed by PJ–SPI and PE–MD. protective effect. When the microcapsules were added to
When compared the encapsulating agent effect on the yogurt the stability of the bioactive compounds followed
polyphenols rate degradation constant, MD showed a a similar behaviour to those without encapsulation
significantly greater protective effect than SPI, for the (juice), except for PE–MD. The microencapsulated
juice systems. The same behaviour showed the antho- anthocyanins disappear before 7 days, according to the
cyanins degradation rate constant for the juice and also observed by Coisson et al. (2005) during the addition of
for the extract system. Contrary, Kim & Morr (1996) açai (Euterpe oleracea) juice to yogurt (10% w ⁄ w) where
determined that SPI shows the lower lost limonene the anthocyanins were stable only 2 days at 4 C. The
release rate on microencapsulated orange oil, respect to anthocyanins degradation rate could be affected by the

International Journal of Food Science and Technology 2010, 45, 1386–1394  2010 Institute of Food Science and Technology
 Encapsulation of polyphenols and anthocyanins P. Robert et al. 1393

Polyphenols retention (%)

160 (a)

Polyphenols retention (%)

160 (b)
120
120

80
80

40 40

0 0
0 20 40 60 0 20 40 60
Time (days) Time (days)

(c) (d)

Anthocyanin retention (%)

160 160

Anthocyanin retention (%)

120 120

80 80
Figure 4 Evolution of the polyphenol and
anthocyanins retention of microcapsules 40 40
obtained under optimal conditions during
storage at 60 C. (a, c) Juice systems and (b, 0 0
0 20 40 60 0 20 40 60
d) extract systems. PJ–MD (d), PJ–SPI ( ),
control ()), PE–MD (s) and PE–SPI (4). Time (days) Time (days)

Table 4 First-order degradation rate constant (k) of polyphenols and anthocyanins from pomegranate microcapsules obtained under optimal
conditions stored at 60 C and added to yogurt

Storage at 60 °C Added to yogurt and storage at 4 °C

Polyphenols Anthocyanins Polyphenols

System 102kobs ± 102SD (days)1) 102kobs ± 102SD (days)1) 103kobs ± 103SD (days)1)

PJ–MD 0.35 ± 0.078a 0.49 ± 0.046a 7.43 ± 2.80b

PJ–SPI 1.36 ± 0.354b 1.98 ± 0.356b 6.22 ± 0.98ab
PE–MD 1.27 ± 0.125b 0.97 ± 0.036c 5.34 ± 0.55a
PE–SPI 1.57 ± 0.040b 2.95 ± 0.091d 7.48 ± 0.65b

PJ, pomegranate juice; PE, pomegranate ethanolic extract; SPI, soybean protein isolates; MD, maltodextrin GAE, gallic acid equivalent,
SD, standard deviation. Different letters show significant differences (P < 0.05).

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International Journal of Food Science and Technology 2010, 45, 1386–1394  2010 Institute of Food Science and Technology