Food Chemistry 187 (2015) 174–181

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Microencapsulation of betalains obtained from cactus fruit

(Opuntia ficus-indica) by spray drying using cactus cladode
mucilage and maltodextrin as encapsulating agents
María Carolina Otálora a, José Gregorio Carriazo b, Laura Iturriaga a, Mónica Azucena Nazareno a,⇑,
Coralia Osorio b,⇑
a
Centro de Investigación y Transferencia de Santiago del Estero (CITSE), National University of Santiago del Estero, CONICET, Santiago del Estero, Argentina
b
Departamento de Química, Universidad Nacional de Colombia, AA 14490 Bogotá, Colombia

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

Article history: The microencapsulation of betalains from cactus fruit by spray drying was evaluated as a stabilization
Received 20 January 2015 strategy for these pigments. The betalains used as active agent were extracted from purple fruits of
Received in revised form 16 April 2015 Opuntia ficus-indica (BE) and encapsulated with maltodextrin and cladode mucilage MD–CM and only
Accepted 20 April 2015
with MD. The microcapsulates were characterized by scanning electron microscopy (SEM), thermal
Available online 23 April 2015
analysis (TGA–DSC), tristimulus colorimetry, as well as, their humidity, water activity and dietary fiber
content were also determined. The active agent content was measured by UV–Vis spectrophotometry
Chemical compounds studied in this article:
and its composition confirmed by HPLC–ESIMS. A pigment storage stability test was performed at
Indicaxanthin (PubChem CID: 6096870)
Betanin (PubChem SID: 10733)
18 °C and different relative humidities. The addition of CM in the formulation increased the encapsulation
Isobetanin (PubChem CID: 6325438) efficiency, diminished the moisture content, and allowed to obtain more uniform size and spherical
particles, with high dietary fiber content. These microencapsulates are promising functional additive to
Keywords: be used as natural colorant in the food industry.
Microencapsulation Ó 2015 Elsevier Ltd. All rights reserved.
Betalain
Nopal
Cactus pear fruit
Natural colorant

1. Introduction extracts of its cladode (modified stems) have shown hypolipi-

Since food ingredients are on the focus of public interest, it is
becoming increasingly important to meet consumers expectations
for natural and healthy products. Hence, the search of new plant-
derived colorants for the food industry is still necessary.
However, the higher stability of synthetic colorants with respect
to natural alternatives is a challenge that must be overcome. In
order to meet the growing demand for natural colorants, new pig-
ment sources are being sought. In this course, red cactus pear
(Opuntia ficus-indica) has been suggested as a valuable source of
betalains, with several technological and sensorial advantages in
comparison to red beet (Moßhammer, Stintzing, & Carle, 2006).
O. ficus-indica (Cactaceae), commonly named as prickly pear or
nopal cactus, develops properly either arid or semiarid lands where
the population uses it as source of food and by foraging. The
⇑ Corresponding authors. Tel.: +54 385 4509500x1617; fax: +54 385 4509525 (M.
Nazareno). Tel.: +57 1 3165000x14452; fax: +57 1 3165220 (C. Osorio).
E-mail addresses: manazar2004@yahoo.com, nazareno@unse.edu.ar
(M.A. Nazareno), cosorior@unal.edu.co (C. Osorio).
demic, hypocholesterolemic, antidiabetic, hypoglycemic, and
anti-inflammatory activities. Additionally, the fruit is rich in
polyphenols and betalains, compounds that are well known by
their antioxidant properties (El-Mostafa et al., 2014; Osuna-
Martínez, Reyes-Esparza, & Rodríguez-Fragoso, 2014). Cactus fruit
is considered a crop widely distributed in America with a fast
growth and low cost, what increases its potential to develop
added-value food products. Cactus betalains have increasingly
attracted interest as a source of water-soluble pigment prepara-
tions (Stintzing, Schieber, & Carle, 2003; Moßhammer et al.,
2006). Depending upon the nature of the betalamic acid addition
residue, the betalains can be divided in two groups: the purple-
red betacyanins and the yellow betaxanthins. The major beta-
cyanin found in cactus pear fruits is betanin that results from the
condensation of betalamic acid with a C5-glucosylated cyclo-Dopa
moiety (Castellanos-Santiago & Yahia, 2008).
Spray drying is the most commonly used encapsulation tech-
nique for food products. The final product quality and powder effi-
ciency depend on the operating conditions, such as, inlet and outlet
air temperatures, feed flow rate, atomization speed or pressure,

http://dx.doi.org/10.1016/j.foodchem.2015.04.090
0308-8146/Ó 2015 Elsevier Ltd. All rights reserved.
 M.C. Otálora et al. / Food Chemistry 187 (2015) 174–181
feed to carrier ratio, among others (Tonon, Brabet, & Hubinger,
2008). Carbohydrates, semi-synthetic derivatives of cellulose, and
synthetic polymers have been used as encapsulating agents,
exhibiting differences in their properties, cost, and encapsulation
efficiency (Davidov-Pardo, Arozarena, & Marín-Arroyo, 2013).
Among them, partially hydrolyzed starch products are the most
widely used additives to obtain fruit juice powders. These white
L-glucose polymers exhibit a neutral taste, are odorless, easily
digested by humans, and classified according to their degree of
hydrolysis, expressed as dextrose equivalent (DE). It has been
reported a significant influence of the dextrose equivalent (DE)
value on the drying yield. The yield increases as well as DE value
of maltodextrin decreases, because the glass transition
temperature is increased.
Some attempts to develop betalain-rich powders from cactus
fruit have been reported by Moßhammer et al. (2006), Saénz,
Tapia, Chávez, and Robert (2009). They concluded that pigment
retention in microcapsules using fruit pulp was higher than that
observed from ethanolic extract as active agent. This protecting
effect was ascribed to the other pulp components such as mucilage,
that plays a significant role during the encapsulating process.
The mucilage extracted from the cladodes of O. ficus-indica is a
promising natural material to be used in food industry due to its
emulsifying properties (Medina-Torres, Brito-De La Fuente,
Torrestiana-Sanchez, & Katthain, 2000). This material has been used
as an edible coating to extend the shelf life of food products (Del
Valle, Hernández Muñoz, & Galotto, 2005). Previous studies have
shown that the mucilage of O. ficus-indica cladode is a
heteropolysaccharide with a molecular weight varying from
2.3  104 to 3  106 gmol1, being a complex mixture of polysaccha-
rides of L-arabinose (24.6–42%), D-galactose (21–40.1%), D-xylose
(22–22.2%), L-rhamnose (7–13.1%), and D-galacturonic acid (8–
12.7%) (Medina-Torres et al., 2000; Sáenz, Sepúlveda, & Matsuhiro,
2004). Several scientific studies report the medicinal properties of
cactus cladode extracts and these beneficial effects have been attrib-
uted to the mucilage action (Nazareno, 2013, 2014).
Betalain stability is affected by several factors such as pH, water
activity, and exposure to oxygen, light, temperature, and degradative
enzymes (Herbach, Stintzing, & Carle, 2006). Microencapsulation has
been proposed as successful strategy to improve the betalain stabi-
lization, to make easier their handling during processing, and to
ensure its bioavailability when they are used as food colorants.
Thus, the aim of this work was to encapsulate the bioactive pigments
of O. ficus-indica fruit pulp by spray drying, using cladode mucilage
and maltodextrin as wall materials in order to obtain betalain micro-
capsules to be used as natural food additives.
2. Materials and methods
2.1. Plant material
Fruits (purple pulp) and cladodes of O. ficus-indica were col-
lected from the cactus collection belonging to the Faculty of
Agronomy and Agroindustries (National University of Santiago
del Estero) at the province of Santiago del Estero, located in the
Argentinean region of the dry Chaco (27°450 S, 64°180 W, 170 m over
sea level). Cladodes were cut in April 2013 and fruits were col-
lected in two different harvesting times (January 2012 and
January 2013). When plant material did arrive to the laboratory,
it was immediately processed to obtain the cladode mucilage
(CM) and the lyophilized pulp (LP).
2.2. Preparation of betalain extract
Cactus fruit pulp was crushed in a food processor and the seeds
removed by filtration. The homogenized cactus fruit pulp was
175
lyophilized in a freeze-dryer Labconco Freezone 4.0 (Kansas City,
MO, USA) to reach a final moisture content between 1.9% and
2.3% (wet base). Then, the sample was kept in hermetic flasks with
protection from light, and stored at 20 °C until use. To obtain
betalain-rich extract (BE), lyophilized cactus pulp was macerated
with a phosphate buffer (pH 5) in ratio 1:5 (w/w).
2.3. Mucilage extraction
Cladodes were transversally cut and manually peeled. The cla-
dode outer layer (epidermis, thorns, and chlorenchyma) was
removed, leaving only the white and inner part called medulla.
The medulla was cut into small slices of 2 mm thickness, and these
slices were put into a stainless steel container with distilled water
in the ratio of 1:2 (medulla/water). The mixture was kept at 20 °C
for 24 h. After pressing the plant material into a self-made
Frautschi extruder (Santiago del Estero, Argentina), a viscous liquid
was obtained and subsequently centrifuged at 10,000 rpm for
30 min. Then, the addition of three volumes of ethanol,
precipitated the mucilage. This white solid was separated and
placed in a forced air convection drying oven at 105 °C for 24 h.
The dried material was ground in a mortar to yield a fine powder,
and then stored at room temperature and relative humidity (RH) of
35% until its use (Quinzio, Corvalán, Lopez, & Iturriaga, 2009).
2.4. Spray-drying process
The betalain-rich extract (BE) was separately combined with
two different carrier agents, maltodextrin DE 20 (MD), and cladode
mucilage (CM). The spray-drying process was performed in a labo-
ratory-scale LabPlant SD-06 spray-drier (LabPlant, Huddersfield,
England), with an 1110  825  600 mm main spray chamber. In
each case, the feed-mixture was kept under constant magnetic
stirring at 20 °C until homogeneity, and spray-dried immediately
with an air flow of 100 m3/h and a compressor air pressure of
4 bar. The nozzle internal diameter was 0.5 mm, feed flow was
485 ml/h, and the inlet and outlet air temperatures were 170 ± 2,
and 98 ± 4 °C, respectively. Thus, two microencapsulates were
obtained: SD1 (BE:MD:CM, 1:1:0.225 w/w/w), and SD2 (BE:MD,
1:1 w/w), collected in plastic containers, weighed, and stored in
desiccators containing silica gel at room temperature.
2.5. Quantitative analyses
2.5.1. Encapsulation yield (EY)
The EY was determined as the percentual ratio between the
total mass of recovered product after spray-drying per the mass
of the extract fed to the system (in dry basis) as it was suggested
by Su et al. (2008).
2.5.2. Betalain content
Microcapsules (100 mg) were accurately weighed and dispersed
in 5 ml of distilled water. This suspension was stirred using a
Vortex for 1 min, and filtered with a Millipore membrane
(0.45 lm). The same procedure was used for BE as control mea-
surements. The betalains were quantified by spectrophotometry
using a Jenway 7305 UV/visible spectrophotometer (Jenway,
Staffordshire, UK) at k 536 nm according to the method reported
by Coria Cayupán, Ochoa, and Nazareno (2011). The data so
obtained were expressed using betanin as reference compound
(e = 65,000 l/mol cm; molar mass = 550.1 g/mol, and k = 536 nm).
2.5.3. Dietary fiber content
Soluble (SDF) and insoluble (IDF) dietary fiber contents were
determined according to AOAC enzymatic–gravimetric method
176 M.C. Otálora et al. / Food Chemistry 187 (2015) 174–181
985.29 (AOAC, 1997). Total dietary fiber (TDF) was calculated as
the sum of SDF and IDF, and expressed as g per 100 g of powder.
2.6. Characterization of the microencapsulates
2.6.1. Moisture content and water activity
Moisture content of the microparticles was determined gravi-
metrically by drying 20 mg of microparticles, in triplicate, at
70 °C until constant weight (AOAC, 1997). The water activity (aw)
of microencapsulates was measured in a hygrometer HygroPalm
AW1 (Rotronic Instruments, Huntington, NY, USA) at 20 °C.
Samples (1 g of each) were analyzed in triplicate and aw value were
reported as the mean of obtained data.
2.6.2. Color measurement
The color of microcapsules was determined by using a
spectrophotometer-colorimeter (HunterLab, ColorQuest XT, USA)
to obtain the CIELAB parameters (L⁄, a⁄, b⁄). The other parameters,
chroma (Cab⁄) and hue (hab), were calculated according to the fol-
lowing Eqs. (1) and (2) (Meléndez-Martínez, Vicario, & Heredia,
2003):
 2
1 25 days). The dependent variables were the water activity (aw), and
C ab ¼ ½ða Þ2 þ ðb Þ 
2
ð1Þ

hab ¼ arctan ðb =a Þ ð2Þ
Euclidean distance between two points in the three-dimen-
sional space defined by L⁄, a⁄, and b⁄ were used for calculating color
differences (DE⁄ab) (Eq. (3)) (Cejudo-Bastante, Hurtado, Mosquera,
& Heredia, 2014):
2  2 1=2
DE ab ¼ ½ðDL Þ þ ðDa Þ2 þ ðDb Þ  ð3Þ
2.6.3. Particle morphology and size
The morphology of microcapsules was evaluated by using a
scanning electron microscope (SEM) FEI QUANTA 200 (operating
at 30 kV), coating the samples by gold sputtering before their
examination. The particle size distribution was determined by
measuring the diameter of every 250 particles localized in a
selected area (ca. 80%) of SEM images.
2.6.4. Thermal analysis
Thermal analyses (TGA/DSC), and glass transition temperature
measurements, were carried out using a simultaneous Analyser
NETZSCH STA 409 CD (Netzsch, Selb, Germany). The apparatus
was calibrated with high purity indium (Tm = 429.8 K,
DHm = 28.4 J g1). The experiments were performed under nitro-
gen flow (50 cm3/min). The samples (2 mg each) were heated from
20 to 350 °C in aluminum crucibles with a linear heating rate of
10 °C/min, and an empty aluminum crucible was used as the refer-
ence material.
2.7. Betalain analysis by HPLC–MS
HPLC–MS analysis of the betalain-rich extract (BE, 3 mg/ml) and
the microencapsulate solutions (10 mg/ml) was performed using a
LCMS–IT-TOF liquid chromatography coupled to a mass spectrom-
eter (Shimadzu, Kyoto, Japan) equipped with an electrospray ion-
ization (ESI) probe, which was operated in positive ion mode. A
C18 column 5 lm (150  4.6 mm i.d., Shimadzu, Japan) was used
for the analysis of the betalains present in each sample. The solvent
system was composed of solvent A, a mixture of water/formic acid
(99:1, v/v), and solvent B, acetonitrile/formic acid (99:1, v/v). The
HPLC program started with a linear gradient from 3% to 14% B in
15 min; flow rate 0.4 ml/min; injection volume of 30 ll; UV–Vis
detection at k 484 and 535 nm, according to that reported by
Fernández-López, Castellar, Obón, and Almela (2002) with some
modifications. The MS/MS parameters were as follows: positive
mode; CDL temperature 200 °C; heating block at 200 °C; detector
voltage, 2.00 kV; flow nebulizing gas (N2), 1.5 L/min; ion accumu-
lation 30 ms; and scan range m/z, 50–1000 u. The energy of the col-
lision gas (argon) was fixed at 50%. The LC–MS Solution software
(Shimadzu, Tokyo, Japan) was used for data collection and analysis.
2.8. Powder storage
SD1 and SD2 samples were weighed (1.5 g of each) in Petri
dishes (2 cm diameter) and stored in the absence of light, at con-
stant temperature (18 ± 1 °C), and at carefully controlled relative
humidities. The samples were placed in sealed desiccators contain-
ing 200 ml of saturated solutions of KNO3, NaCl, and NaBr, to
obtain constant humidity values of 90 ± 2%, 75 ± 2%, and 57 ± 2%,
respectively. During equilibration, the humidity was measured
using a thermohygrometer (model 445815; Extech Instruments,
Nashua, NH, USA). The experimental design was a 2  3  7 com-
plete factorial in a random design. The independent variables con-
sidered were humidity and time of storage (2, 4, 7, 10, 15, 20 and
betalain content (see Section 2.5.2), which were determined by
triplicate removing samples after each measurement. The observed
first-order release rate constant (k) and half-life time (t1/2) were
calculated by the method reported by Cai and Corke (2000). Half-
life (t1/2) for the retention of betanin was calculated from the rate
constant as 0.693/k.
The pigment retention was determined by spectrophotometric
method as it was described in Section 2.5.2, and calculated accord-
ing to Eq. (4).
Mt
Betanin retention ð%Þ ¼  100 ð4Þ
M0
where Mt and M0 are the absolute amounts of betanin dispersed in
distilled water at storage time and before starting the experiment
(zero storage time), respectively.
2.9. Statistical analysis
Data from the microencapsulate characterization were reported
as the mean ± standard deviation for determinations performed
(n = 3). Using the statistical package SAS 9.4 (SAS Institute Inc.,
Cary, NC), analysis of variance (ANOVA) was performed to identify
differences among the means. Differences at probability level
p 6 0.05 were considered significant.
3. Results and discussion
3.1. Characterization of betalain-rich microencapsulates
Two betalain-rich microencapsulates (SD1 and SD2) were
obtained from cactus fruit using maltodextrin as encapsulating
agent, but cladode mucilage (CM) was added to one of them
(SD1) to benefit from biofunctional and nutritional properties of
this promising natural material. The amount of CM added to SD1
was determined according to its low water solubility reported by
León-Martínez, Rodríguez-Ramírez, Medina-Torres, Méndez-
Lagunas, and Bernad-Bernad (2011) as 1–6% (w/v). The character-
ization results for SD1 and SD2 are shown in Table 1. Water activity
(aw) values were 0.176 for SD1 and 0.205 for SD2. These values are
below the maximum accepted (0.600) to prevent microbial decom-
position of foods. The process to obtain SD1 had the highest yield,
attributable to the presence of CM that improved the spray-drying
process; which is in good agreement with the lower water activity
 M.C. Otálora et al. / Food Chemistry 187 (2015) 174–181 177

Table 1
Yield of encapsulation, water activity (aw), moisture, betalain content, color parameters, and dietary fiber content of cactus microencapsulates, cactus fruit pulp and cladode
mucilage.

Parameter Sample
SD1 SD2 Cactus fruit pulp Cladode mucilage (CM)
EY (%) 25.4 16.4 – –
aw at 18 °C 0.176 ± 0.001a 0.205 ± 0.001b – –
Moisture (%) 2.9 ± 0.2a 4.8 ± 0.8b – –
Betalain content* 0.497 ± 0.003a 0.495 ± 0.005a – –
Total dietary fiber (TDF)** 4.42 ± 0.1a 3.44 ± 0.1b 4.15 ± 0.1 57.70 ± 1.1
Soluble dietary fiber (SDF)** 1.41 ± 0.1a 1.22 ± 0.3b 1.64 ± 0.2 27.29 ± 0.1
Insoluble dietary fiber (IDF) and percentage** 3.01 ± 0.1a (68.1%) 2.22 ± 0.9b (64.5%) 2.51 ± 0.3 (60.5%) 30.41 ± 1.0 (52.7%)
Color parameters
L* 66.46 ± 0.12a 66.04 ± 0.24a – –
a* 29.99 ± 0.13a 30.14 ± 0.13a – –
b* 4.84 ± 0.05a 5.28 ± 0.07b – –
Cab* 30.37 ± 0.02a 30.59 ± 0.01a – –
hab 9.16 ± 0.01a 9.93 ± 0.01b – –

All data are the mean of measurements (n = 3) ± standard deviation. Different letters in the same column indicate a statistical difference (p < 0.05).
–, not recorded.
*
Expressed as mg betanin equivalents/g solid.
**
Expressed as g/100 g.

(aw) and moisture values obtained for SD1 in comparison to SD2. Table 2
The presence of CM in the formulation of SD1, decreases the adhe- Chromatographic and spectrometric data (HRESIMS) of the BE and betalain-rich
microencapsulate solutions from cactus fruit (Opuntia ficus-indica).
sion of product into the drying chamber, thus increasing the encap-
sulation yield. This tendency has been found in other studies Peak Compound tR kmax Precursor ion MS2
dealing with the recovery of powders produced by spray-drying (min) (nm) (m/z)

using mucilages (León-Martínez, Méndez-Lagunas, & Rodríguez- 1 Indicaxanthin 5.25 484 309.0540 262.9774/
Ramírez, 2010; Cervantes-Martínez et al., 2014). For mucilaginous 221.0480
2 Betanin 6.12 535 550.8038 388.9305
materials with high sugar content, thermoplastic behavior is also 3 Isobetanin 7.50 535 550.8240 –
affected by temperature, resulting in low yields (Gharsallaoui,
Roudaut, Chambin, Voilley, & Saurel, 2007); however, the temper- –, not recorded.

ature values used in the present study did not affect the yields.
High temperatures (>170 °C) promote changes in both, the struc-
ture and thermoplastic phase of the materials, such increasing
deposits on the dryer walls, favoring the degradation of the molec-
ular mucilage structure, and diminishing viscosity (León-Martínez
et al., 2010).
3.2. Color analysis
respectively. The presence of betanin (peak 2) and its 15-R-epimer,
isobetanin (peak 3), was confirmed by their pseudomolecular ions
[M+H]+ at m/z 550.8038 and 550.8240, respectively. During MS/MS
analysis, pseudomolecular ion of peak 2 produced the fragment ion
at m/z 388.9305 corresponding to the loss of hexose moiety
(162 u).

The color obtained can be described as a vivid red–purple
resembling a deep radish tone (Cai & Corke, 2000). The projection
of the data onto the a⁄, b⁄ diagrams showed that all data are located
in the second quadrant (+a⁄, b⁄). No differences in L⁄, a⁄ and C⁄ab
values were found for both samples. Taking into account that
DE⁄ab of up to two CIELAB units indicates color differences appre-
ciable to the human eyes (Cejudo-Bastante et al., 2014), it was con-
firmed that there were no color differences visually appreciable
among two powders because this value was 0.627. The betalain
contents in SD1 and SD2 microcapsules were 0.497 ± 0.03 y
0.495 ± 0.05 mg betanin equivalents/g of beads respectively, show-
ing that the type of encapsulating agent did not affect the retention
of the bioactive compound a time zero, in agreement with data of
color measurement. This conclusion was confirmed by HPLC–MS
analyses of the microencapsulate solutions (Table 2). The betalain
composition of microcapsules SD1 and SD2 was qualitatively sim-
ilar to the BE betalain profile measured at k 484 and 535 nm, show-
ing three betalains peaks in agreement with those results
published by Fernández-López et al. (2002). Peak 1 was identified
as indicaxanthin by the presence of the pseudomolecular ion at
m/z 309.0540 [M+H]+ and the ion fragments at 262.9774 and
221.0480 obtained by tandem mass spectrometry (MSn) with an
ion trap when the pseudomolecular ion was selected as precursor,
explained as [M+H–COOH]+ and [M+H–CH2CH(COOH)NH]+,
3.3. Particle size and morphology
Cactus pear lyophilized pulp is shown in Fig. 1A; the image
reveals ‘‘amorphous’’ matrices with internal cavities (Habibi,
Mahrouz, & Vignon, 2009), possibly due to the lyophilization pro-
cess. Fig. 1B shows MD particles being very irregular in form and
size. The particle morphology of CM powders obtained by drying
in a forced air convection oven is shown in Fig. 1C. The image anal-
ysis reveals a spherical and non-agglomerated particles with uni-
form appearance, suggesting an uniform drying. The spherical
nature of the particles allows powder to flow better. According to
Walton and Mumford (1999) those spherical particles can flow
freely because they lack surface roughness and do not form agglom-
erates. Additionally, agglomeration of the particles depends on the
properties of the material, and may be reduced by modifying drying
conditions (García-Cruz, Rodríguez-Ramírez, Méndez-Lagunas, &
Medina-Torres, 2013). The spherical morphology of CM shows the
favorable tendency of this wall material to yield microcapsules.
This is likely a result of the high surface tension of the molecular
chains contained in the cactus cladode mucilage after dehydration,
which reveals their physicochemical potentiality for encapsulating
pigments and bioactive molecules by spray-drying.
SEM micrographs confirm the successful formation of micro-
capsules for both SD1 and SD2 spray-dried powders. A careful
178 M.C. Otálora et al. / Food Chemistry 187 (2015) 174–181
Fig. 1. Scanning electron micrographs of the starting materials and microcapsules.
Cactus pear lyophilized pulp (A), maltodextrin DE 20 (MD) (B), cactus cladode
mucilage (CM) (C), betalain-rich microcapsulates SD1 (D and E), and SD2 (F and G).
comparison of the morphologies observed for the microencapsu-
lated materials by spray-drying (Fig. 1D–G) shows that the SD1
microcapsules (Fig. 1D and E) were more spherical than those of
SD2, with smooth surface and very low agglomeration level. This
is probably due to the high molecular weight of wall material
(CM) that prevents surface contraction, allowing the formation of
a continuous matrix. The SD2 microcapsules (Fig. 1F and G)
frequently showed dented surfaces with roughness and higher
agglomeration of particles. This morphology may be attributed to
the shrinkage of the particles during the drying process (Medina-
Torres et al., 2013), which is a consequence of the presence of
maltodextrin. Similar morphology was observed in microcapsules
of Opuntia lasiacantha pigments with maltodextrin (Díaz, Santos,
Kerstupp, Villagómez, & Scheivar, 2006). Maltodextrin content
provides low-molecular weight sugars that can act as plasticizers,
which reduce the polymer chain contacts, thereby decreasing the
rigidity of the three-dimensional film structure (Villacrez,
Carriazo, & Osorio, 2014). The more plastic behavior of these
materials explains the frequent appearance of floppy film and
rough wall on the SD2 microcapsules.
The size of atomization nozzle of 2 mm allowed obtaining fine
droplets sprayed into the drying chamber producing uniform
particles with defined morphology. The particles of SD1 powder
have mainly ranged from 6 to 30 lm in size; this indicates that
the CM addition allows obtaining fine particles as reported by
León-Martínez et al. (2010). The average particle size of the SD2
microcapsules ranged from 6 to 51 lm. The SD1 powder has a
more homogenous distribution, such confirming that the
interaction MD–CM led to a better control on the size of particles.
Evidently the particle size of SD1 was smaller than that of SD2.
It is important to note that the increase on particle size is related to
the molecular size of the carrying agent (Tonon, Brabet, &
Hubinger, 2010). On the other hand, according to the literature,
MD may induce rapid formation of a vitreous surface, which allows
the expansion of air in the interior of the particles, such favoring an
increase in particle diameter (Pierucci, Andrade, Farina, Pedrosa, &
Rocha-Lehao, 2007).
3.4. Thermal analyses of microencapsulates
Thermal analysis is a suitable technique to obtain information
on drying, dehydration, dehydroxylation, gelatinization, melting
and glass transition occurred in food materials when they are
subjected to thermal changes. Simultaneous thermal analyses
(TGA–DSC) of cactus cladode mucilage (CM), maltodextrin (MD),
and the microencapsulated powders (SD1 and SD2) are shown in
Fig. 2. In all cases a relative thermal stability was observed up to
100 °C, with a weight loss below 7%. However, MD is more stable
than CM, SD1, and SD2. This important behavior is very useful
for several food industrial operations. Three general thermal events
were observed for the thermogravimetric curves (TGA), the first
below 200 °C corresponding to the loss of water contained in the
materials, and the others events likely are related to the loss of
mass because of thermal dehydroxylation/decomposition and
volatilization of melted materials. On the other hand, DSC curves
show complex endothermic behavior for all analyzed materials.
These curves provide the glass transition (Tg) values, revealing
45 °C for CM and 52 °C for MD, which are in agreement with the
those data reported in literature (León-Martínez et al., 2010;
Fazaeli, Emam-Djomeh, Ashtari, & Omid, 2012). Tg values observed
for both SD1 and SD2 powders were about 40 °C. These values
result from different interaction between the betalain extract
(BE) components and the carrying materials. Tg is related to molec-
ular weight of species contained in the materials, but also to chain
stiffness and polymer chain structure; Tg increases as cross-link
density increases (León-Martínez et al., 2010). Both mucilage and
maltodextrin are different types of polysaccharides, whose struc-
tures are able to form interactions such as dipole–dipole and
hydrogen bonding with molecules like betalains and water. These
interactions cause disruption in polymeric aggregations, modifying
the size of molecular aggregates of wall materials, and yielding
slight variation in the glass transition temperatures.
The complex and broad endothermic signals observed in DSC
curves of the samples is a consequence of successive thermal
events related to softening and fluidizing of samples as result of
their gelatinisation and melting. The broad endothermic band
about 100 °C can be attributed to the gelatinization process. In this
process, starch, carbohydrate chains, and similar structures are
decomposed in presence of water when they are heated. This
decomposition is accompanied with both phenomena, a higher
molecular disorder and heat absorption. However, DSC profiles of
these materials are very complex because the water evaporation
(an endothermic event) is also carried out throughout the heating
of samples. The broad endothermic peak around 200 °C is related
to the complete melting of materials and possibly partial evapora-
tion of the liquids. Evaporation is considered due to the strong
dropping of mass (TGA curve) immediately after the above
mentioned endothermic peak. Finally, no relevant differences were
observed from TGA–DSC profiles obtained for SD1 and SD2
 M.C. Otálora et al. / Food Chemistry 187 (2015) 174–181
Fig. 2. TGA–DSC profiles for the starting materials and microencapsulated powders: cactus cladode mucilage (CM) (A), maltodextrin DE 20 (MD) (B), microencapsulated SD1
(C), and microencapsulated SD2 (D). Glass transition temperature (Tg) is indicated in each graph.
microencapsulates, which indicates that from a thermal point of
view, these two powders are comparables.
3.5. Dietary fiber measurement
Total dietary fiber (TDF), soluble dietary fiber (SDF) and insol-
uble dietary fiber (IDF) were calculated according to the 985.29
AOAC (1997) method (Table 1). Cactus pear fruit pulp is composed
by lignin, cellulose, hemicellulose, pectin, gum and mucilage, min-
erals and natural antioxidant compounds (Feugang, Konarski, Zou,
Stintzing, & Zou, 2006). TDF content is ca. 4% approx., with a high
insoluble fiber content, consistent with the data published by Díaz
Medina, Rodríguez Rodríguez, and Díaz Romero (2007). Álvarez
and Peña-Valdivia (2009) reported that the distribution of fiber
fractions varies according to the stage of maturation; in particular,
mucilage and pectin content increased during the fruit ripening.
Furthermore, O. ficus-indica cladodes exhibited a high content of
TDF, being IDF more than 50% of TDF. Despite O. ficus-indica cla-
dode mucilage is recognized to be rich in dietary fiber, reports con-
cerning dietary fiber composition in CM are still scarce. Dietary
fiber is a nutritional component associated with some biofunc-
tional properties such the decreasing of cholesterol levels in the
blood, the prevention and control of diabetes due to the control
of glucose (Osuna-Martínez et al., 2014), and reducing symptoms
of chronic constipation, diverticular disease, and hemorrhoids,
among other diseases (Sáenz et al., 2004). Insoluble dietary fiber
percentage increased for all microencapsulates in relation to the
cactus fruit pulp and CM, being higher in SD1 than in SD2. Thus,
the presence of SDF in the CM gives an added-value to the
microencapsulate SD1, making this powder useful not only as col-
orant but also as health-promoting natural additive.
179
3.6. Storage stability test
The water activity (aw) of SD1 and SD2 betalain-rich microen-
capsulates was measured during 25 days of storage at different
RH (Fig. 3A). In general, the aw values tended to be constant after
7 days, obtaining similar values for both powders. This result appar-
ently indicates that the amount of CM in SD1 does not modify the
storage stability (at a fixed relative humidity, RH) in comparison
with the SD2. Therefore, this comparable behavior among SD1
and SD2 reveals that CM is a successful encapsulating material.
On the other hand, the betanin retention during storage was
compared for SD1 and SD2. The pigment retention in SD1 at low
humidity value (57% RH), after 25 days storage was 75.3%, in con-
trast, at higher humidity (90% RH), the pigment retention
decreased until 25.9% with respect to the initial concentration of
the betanin (Fig. 3B). Similar behavior was found for SD2 sample,
allowing to evidence a strong influence of the relative humidity
on the storage stability pigment.
In SD1, the presence of CM decreased the moisture content. The
lower moisture content of the powders ensures the stability of the
product; however, microcapsulates with low moisture content are
more hygroscopic, so, they have a greater capacity to absorb
humidity from environment, what is related to the increase of
water concentration gradient in the product and from the sur-
rounding air (Tonon, Brabet, Pallet, Brat, & Hubinger, 2009). In
SD2, the higher content of maltodextrin DE-20 causes an increase
in powder moisture content, because lower molecular weight mal-
todextrins contain shorter chains and more hydrophilic groups (Cai
& Corke, 2000).
In SD2 sample, high humidity conditions (90% RH) reduced the
pigment retention at 43.3%, compared to 89.2% obtained at lower
180 M.C. Otálora et al. / Food Chemistry 187 (2015) 174–181

1.0 Table 3
A Betanin degradation rate constants of cactus fruit betalain-rich microcapsules during
0.90 their storage at different relative humidity (RH) and constant temperature (18 °C).
Water acvity (Aw)

90% RH Microencapsulate Treatment kobsd Correlation Half-life

0.80
75% RH (RH) (%) (days1) coefficient (r) period t1/2
57% RH
(days)
0.70
90% RH SD1 90 0.049 0.962 13.9
0.60 75% RH
75 0.013 0.960 51.7
57 0.012 0.973 54.1
57% RH
0.50 SD2 90 0.029 0.926 23.8
75 0.007 0.958 103.4
0.40 57 0.006 0.964 117.4
0 5 10 15 20 25
Time (days)
100 rate constant of pigment degradation for both samples, being more
90
B noticeable for SD1 sample at 90% RH (Table 3). The results clearly
Betanin retenon (%)

showed that the stability of encapsulated betalains during storage

80 95% RH
90% RH
depended on both the % RH value and the type of encapsulating
70 75% RH
agent.
60 57% RH
From Fig. 3C it is possible to find differences in the degradation
50 95% RH
90% RH kinetics of SD1 and SD2. SD2 powder was more stable than SD1 at
40 75% RH 57 and 75% RH, with half-life period of 117.4 and 103.4 days,
30 57% RH respectively. It seems that pigments in both samples are easily
20 degradated at 90% RH, with the lowest value for SD1 (t1/2 decreased
0 5 10 15 20 25 to 13.9 days). This can be ascribed to the low protection provided
Time (days) by the matrices to the active agent as consequence of their hygro-
scopicity. Thus, the presence of water enhanced the diffusion of
4.7 betalain molecules through the encapsulating matrix to the cap-
C sule surface.
Ln (% betanin retenon)

4.5

4.3
90% RH 4. Conclusions
4.1 75% RH
57% RH Two microencapsulates were obtained by spray-drying of beta-
3.9
90% RH lain-rich extract and maltodextrin low dextrose equivalent and cla-
3.7 75% RH dode mucilage (CM) from O. ficus-indica with a high recovery of
3.5 57% RH betalains, showing a pigment retention higher than 70% at 18 °C
3.3 and humidity below 57% RH. The most remarkable result was the
0 5 10 15 20 25 stability of O. ficus-indica betalains, under above-mentioned condi-
Time (days) tions. The presence of CM in SD1 significantly increases the TDF
(total dietary fiber) content such giving an added-value to this pur-
Fig. 3. Water activity (aw) variation (A), percentage of betanin retention-vs-time
ple natural colorant as biofunctional additive for food industry.
(B), and first-order kinetic plots for changes in betanin retention (C), during storage
stability test of betalain-rich microencapsulates at 18 °C and different relative Thus, SD1 formulation containing the CM new material confirms
humidity (RH) conditions. Full and empty markers correspond to SD1 and SD2, the great potentiality to use this waste material as encapsulating
respectively. agent in spray drying processes for the food industry.

humidity (57% RH) (Fig. 3B). These results may be explained by the
hygroscopicity of the powder increased at higher humidity.
The SD2 sample could form a more dense and more oxygen
impermeable wall system providing better storage stability for pig-
ments in comparison to the SD1 carrier agents (CM:MD). The
results may be explained by the hygroscopicity of the carrier
agents. Furthermore, these findings suggest that the betanin loss
is increased at high RH due to the capsule wall degradation and
its consequent increase in oxygen permeability. The moisture
adsorbed by the microcapsules produces a decrease in wall density
and opening the pore network, such acceleration this process (Cai
& Corke, 2000). This wall damage results in a faster degradation of
the pigment by oxidation mechanism, which produces the bleach-
ing of the pigment without the appearance of any other absorption
band in the visible spectrum (Attoe & von Elbe, 1985).
The process of pigment stability throughout time for the two
microencapsulates was adjusted to a first order equation according
to the literature (Cai & Corke, 2000; Saénz et al., 2009), and the
results were plotted as ln (% betanin retention) versus t in
Fig. 3C. In general, an increase in storage RH led to an increase in
Acknowledgments
Authors greatly acknowledge the financial support provided by
COLCIENCIAS – Colombia (Contrato RC. No. 707-2013) and
MINCyT, Argentina. M.C.O. acknowledges CONICET for her doctoral
fellowship.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.foodchem.2015.
04.090.
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