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TECNOCIENCIA CHIHUAHUA, Vol. XVIII (4): e1731 (2025)
https://vocero.uach.mx/index.php/tecnociencia
ISSN-e: 2683-3360
Artículo Científico
Encapsulation of betalains from beetroot stem by
spray drying using Aloe vera gel as carrier
Encapsulación de betalaínas de tallo de betabel mediante secado por
aspersión utilizando como agente acarrareador gel de Aloe vera
*Correspondencia: mruizg@uach.mx (Martha Graciela Ruiz-Gutiérrez)
DOI: https://doi.org/10.54167/tch.v18i4.1731
Recibido: 04 de noviembre de 2024; Aceptado: 28 de enero de 2025
Publicado por la Universidad Autónoma de Chihuahua, a través de la Dirección de Investigación y Posgrado.
Editor de Sección: Dr. Gerardo Méndez-Zamora
Abstract
The effect of Aloe vera (A. vera) as a carrier on the encapsulation of beetroot stem juice by spray drying
was studied. Mixtures in % A = A. vera and % B = beetroot stem juice were dried by spray: a) 25A:75B,
150 ºC, b) 25A:75B, 180 ºC, c) 75A:25B, 150 ºC, d) 75A:25B, 180 ºC. The obtained powders were
evaluated for water activity (aw), moisture (M), pH, glass transition temperature (Tg), bulk density
(BD), water solubility index (WSI), color parameters, betacyanin (BC) and betaxanthin (BX) contents,
antioxidant activity (AA) and morphology. The results show that with increasing A. vera H, pH, BD,
Tg, WSI, BC, BX and the tendency to red decreased, while brightness and tendency to yellow
increased and more compressed capsules were observed in the powders. Temperature affected WSI,
color parameters a* and b* and individual betacyanins. The beetroot stem powders presented
characteristics that allow stability: low aw (<0.13), low moisture (<10 %), adequate pH for betalains
(5.3-5.8) and Tg suitable for storage (around 50 ºC). The powder obtained with 25 % A. vera at 180 ºC
presented a higher WSI and a tendency to red color, due to its higher BC and BX content, representing
an alternative as a natural pigment in food formulation.
Keywords: betalains, spray drying, encapsulation, Aloe vera, beetroot stem
José Carlos Castillo-Altamirano1, Miguel Ángel Sánchez-Madrigal1, Armando Quintero-
Ramos1 and Martha Graciela Ruiz-Gutiérrez1*
1 Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua. Circuito Universitario s/n
Campus Universitario 2, Chihuahua 31125, México.
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Resumen
Se estudió el efecto del Aloe vera (A. vera) como acarreador en la encapsulación de jugo de tallo de
betabel mediante secado por aspersión. Mezclas en % A = A. vera y % B = jugo de tallo de betabel
fueron secadas por aspersión: a) 25A:75B, 150 ºC, b) 25A:75B, 180 ºC, c) 75A:25B, 150 ºC, d) 75A:25B,
180 ºC. Los polvos obtenidos se evaluaron para la actividad de agua (aw), humedad (H), pH,
temperatura de transición vítrea (Tg), densidad aparente (DA), índice de solubilidad en agua (ISA),
parámetros de color, contenidos de betacianinas (BC) y betaxantinas (BX), actividad antioxidante
(AA) y morfología. Los resultados muestran que el aumento de A. vera disminuye H, pH, BD, Tg, ISA,
BC, BX y la tendencia al rojo, mientras que la luminosidad y la tendencia al amarillo aumentaron y
se observaron cápsulas más comprimidas en los polvos. La temperatura afectó la ISA, los parámetros
de color a* y b*, y las betacianinas individuales. Los polvos de tallo de betabel presentaron
características que permiten la estabilidad: baja aw (<0.13), baja humedad (<10 %), pH adecuado para
betalaínas (5.3-5.8) y Tg apta para almacenamiento (alrededor de 50 ºC). El polvo obtenido con 25 %
de A. vera a 180 ºC presentó un mayor ISA y tendencia al color rojo, debido a su mayor contenido BC,
representando una alternativa como pigmento natural en la formulación de alimentos.
Palabras clave: betalaínas, secado por aspersión, encapsulación, Aloe vera, tallo de betabel
1. Introduction
Beetroot (Beta vulgaris) has been catalogued among the ten important vegetables (Kavalcová
et al., 2015). The intense red color of beetroot is due to high concentrations of betalains, and these are
one of the compounds that provide beetroot with its antioxidant capacity (García et al., 2016).
Betalains from beetroot, are one of the most widely used natural colorants in the food industry as
the red color is used in many food products. Of the beetroot plant, the main root or bulb is the part
that is consumed; however, the stem contains betalains just like the bulb but it is not consumed
directly, but can be considered as a by-product. The current trend is to recover valuable components
of food by-products and recycle them within the food chain (Galanakis, 2015). The stem is a source
of betalains (Slatnar et al., 2015; Aguirre-Calvo et al., 2018), with the potential to be used as a natural
coloring alternative, the above with the purpose of formulating functional foods or to contribute to
the new objectives in the food industry, such as obtaining clean label. Betalains from beet stems are
a safe and healthy alternative of natural pigment, this approach as a current need.
Betalains are a group of water-soluble nitrogenous pigments with two structural groups: the red-
violet group named betacyanins and the yellow-orange group named betaxanthins (Hussain et al.,
2018). Betalains have also received much attention due to their benefits for human health, especially
for their antioxidant and anti-inflammatory activities among others (García et al., 2016); help in the
treatment of some pathologies (Clifford et al., 2015). But despite its attractive color and health benefits
there is a problem of stability of natural pigments when they are extracted from their sources, being
highly sensitive to processing and storage conditions. To reduce the interactions of these compounds
with factors to which they are sensitive, such as oxygen, humidity, light and the action of
temperature and thereby increase their stability and shelf life, material packaging technologies can
be used as a method of encapsulation. One technique for protecting and improving the stability of
natural pigments is encapsulation by spray drying. This method has several advantages, including
low cost and easy scalability. Even though spray drying has many advantages, the process is carried
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out at high temperature (>100 ºC) and it has been reported that temperature is one of the process
conditions that can cause degradation of components such as betalains, specifically the betacyanins
(Ruiz-Gutiérrez et al., 2014). A key point in the spray drying, to carry out the encapsulation it is
necessary to use an adjuvant or carrier agent in the process. The most commonly used encapsulating
agents are maltodextrin, gum arabic, and modified starch (Sánchez-Madrigal et al., 2019), but in the
search for functional carriers, other types of compounds such as beta-glucans (Ruiz-Gutiérrez et al.,
2014), inulin (Araujo-Díaz et al., 2017) and fructans (Sánchez-Madrigal et al., 2019) have been used,
and more recently, matrices such as the use of maltodextrin-trehalose (Millinia et al., 2024), whey
protein isolate (Wu et al., 2021) and ternary combinations of modified corn starch, gelatin,
maltodextrin, tamarind gum, pectin, inulin and β-cyclodextrin (Guo et al., 2020) have been tested, to
develop encapsulated powders with suitable properties for use as food ingredients.
A. vera gel is increasingly used in food products (Ray et al., 2013; Rodríguez-Rodríguez et al., 2020).
Although the gel contains between 98.5 % and 99.5 % water, the solids are a combination of mucilage
and other carbohydrates, organic acids and salts, enzymes, saponins, tannins, phenolic
anthraquinones, flavonoids, flavanols, sterols, triacylglycerides, amino acids, vitamins, and minerals
(Rodríguez-Rodríguez et al., 2020). This rich composition of functional compounds gives the gel a
wide variety of biological activities, a bioactivity that can be used in ingredients for food formulation.
The objective of this study was to evaluate the use of A. vera gel as a carrier agent for encapsulation
by spray drying of beetroot stem juice, and to study the physicochemical properties of the
encapsulated powder obtained at different temperatures.
2. Materials and Methods
2.1 Materials
Beetroot Stem Juice. Whole beetroot plant (Beta vulgaris) was purchased from a local market
in Chihuahua, Mexico. The stems were removed from the bulbs, then the stems were washed and
disinfected [chlorine solution, 150 ppm] and processed through a domestic juice extractor. The juice
obtained was vacuum filtered by passing the juice through of filter paper (No. 1; Whatman, Kent,
UK). The juice was analyzed for moisture, pH, water activity, the total soluble solids (ºBx),
antioxidant activity, and betalains (betacyanins and betaxanthins) content. The filtered juice was
frozen and stored at −20 ºC in the dark until processing.
Aloe vera gel. A. vera (Aloe barbadensis Miller) was purchased from the greenhouse of the Faculty of
Chemical Sciences of the Autonomous University of Chihuahua. The A. vera leaves were selected for
their size (40 to 50 cm), light gray green color and freshness. The leaves were washed and disinfected
[chlorine solution, 150 ppm]. To obtain the gel, the acibar was first extracted by cutting the bases of
the leaves and allowing them to drain vertically for one hour. Then, the epidermis was removed and
the pulp obtained from A. vera gel was rinsed with distilled water to remove the residue of the acibar
on the surface, processed with a domestic blender and the juice obtained was used immediately.
Some characteristics such as pH, soluble solids, color parameters and antioxidant activity were
determined.
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2.2 Methods
2.2.1 Encapsulation of beetroot stem juice
Mixture: Mixtures of 450 mL of A. vera (% A) and beet stem juice (% B) were prepared 25A:75B
and 75A:25B. The beetroot stem juice was prepared with maltodextrin [30 % (w/w, based on total
soluble solids)] to increase the yield of the encapsulated powder obtained, due to the high-water
content of the A. vera gel. Each mixture was replicated and was passed through a spray dryer keep
with constant agitation.
Spray Drying: A spray dryer (Niro Mobile Minor DK-2860; GEA Company) with a rotary atomizer
at 30 psi air pressure was used. The mixtures were fed through a peristaltic pump (Watson Marlow
504-U; Falmouth, Cornwall, UK) and dried at 150 and 180 ºC for the inlet temperature and 70 and 85
ºC for the outlet temperature, respectively. Spray drying process was performed in duplicate.
2.2.2 Analyses methods:
pH: pH was determined using a pH meter (HANNA Instruments, model EDFE HI2020, RI,
USA) calibrated with standard pH 7.0 and 4.0 buffers. For encapsulated powders, the pH was
determined using the method described by Kha et al. (2010). Encapsulated powder (5 g) was mixed
in 25 mL of deionized water at 20 ºC. The determinations were performed in triplicate and the mean
values with standard deviation were reported.
Total soluble solids: Total soluble solids were measured with the Abbe refractometer (Atago Co. Ltd.,
Tokyo, Japan) which was previously calibrated with sucrose solutions. The determinations were
performed in triplicate and the mean values with standard deviation were reported.
Color: The color parameters of the luminosity (L*), green/red (-/+a*) and blue/yellow (-/+b*) were
recorded samples were performed using Konica Minolta CR-400/410 colorimeter, calibrated with
standards with values X= 94.9, y= 0.3185 and x= 0.3124. The determinations were made in triplicate
and the mean values with standard deviation were reported.
Moisture: The moisture content was determined by gravimetric method 950.02 (AOAC., 1998). The
analysis was carried out in triplicate and the mean values with standard deviation were reported.
The results were expressed as g of water/100 g of encapsulated powder.
Water activity (aw): The aw was measured using a water activity meter (AquaLab Series 3 Quick Start;
Decagon Devices Inc., Pullman, WA, USA). The analysis was performed in triplicate and the mean
values with standard deviation were reported.
Bulk density (BD): The BD was determined using the method described by Kha et al. (2010).
Encapsulated powder (2 g) was placed in a 10 mL graduated cylinder and the cylinder was held in a
vortex vibrator (Vortex-1; Scientific Industries Inc., Bohemia, NY, USA) for 1 min. The BD value was
calculated as the ratio of the mass in the cylinder to the volume occupied in the cylinder. The analysis
was performed in triplicate and the mean values with standard deviation were reported.
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Morphology. Capsules were observed at ×750 magnification with a scanning electron microscope
(JSM-5800 LV; JEOL, Tokyo, Japan). The encapsulated powder was fixed on pieces of double-sided
tape and coated with a thin layer of gold (-110 Å) using a sputter coating vacuum (Desk II; Denton
vacuum LLC., Moorestown, NJ, USA). The samples were then examined by SEM at 10 kV.
Water Solubility Index (WSI). The WSI was determined according to Anderson et al. (1969).
Encapsulated powder (2.5 g) and 30 mL of deionized water were mixed vigorously using a vortexer
(Vortex-1; Scientific Industries Inc.) and incubated in a 37 ºC water bath for 30 min. The mixtures
were centrifuged at 3600 x g for 60 min (Centra Centrifuge CL3R IEC; Thermo Electron Corporation,
Waltham, MA, USA). The supernatant was carefully collected in trays previously weighed to
constant weight and dried (Shel Lab; Sheldon Manufacturing, Inc., Cornelius, OR, USA) at 70 ºC for
36 h. The WSI was calculated as the percentage of the weight of the dried supernatant compared to
the pre-dried weight of the powder. The analysis was performed in triplicate and the mean values
with standard deviation were reported.
Glass transition temperature (Tg): The Tg was measured according to the described method by Ahmed
et al. (2010) using a differential scanning calorimeter (TA Q-200; TA Instruments, Crawley, UK).
Encapsulated powder (8 mg) was placed in an aluminum dish and sealed. An empty sealed capsule
was used as a reference. The temperature scanning program changed a range of 70 - 120 ºC, followed
by cooling to 30 ºC at a temperature ramp of 10 ºC/min under a nitrogen gas atmosphere.
Thermograms were analyzed using universal analysis software (TA Instruments). The glass
transition temperature was interpreted as the midpoint of the curves obtained. The analysis was
performed in duplicate and the mean values with standard deviation were reported.
Betalains (BT) content: Both pigments, betacyanins and betaxanthins contents, were determined by
spectrophotometric method. Beetroot stem juice was diluted with distilled water and the
measurement was performed at a wavelength of 535 and 483 nm. The contents were expressed in
mg/L of betacyanins and betaxanthins, respectively; using the following equation (Castellar et al.,
2003):
󰇛
󰇜 󰇛󰇜󰇛󰇜󰇛󰇜󰇛󰇜
󰇛󰇜󰇛󰇜 󰇛󰇜
Ec. (1)
For encapsulated powders, the pigments were extracted according to the method of Castellanos-
Santiago and Yahia (2008) with some modifications. Encapsulated powders (100 mg) were stirred in
10 mL of deionized water for 15 s using an Ultra-Turrax IKA T18 basic with an S18N19G
homogenizer-dispersing tool (IKA Works Inc.). The samples were then centrifuged at 3,600 × g at 10
ºC for 40 min (Centra centrifuge CL3R IEC; Thermo Electron Corporation). This procedure was
repeated to ensure efficient extraction. The supernatants were filtered through a 0.45 µm nylon filter
(Millipore Corp.). Extractions were performed in triplicate and analyzed by spectrophotometry
(Perkin-Elmer) and identification and degradation (%) by HPLC (1200 series; Agilent Technologies).
The photometric quantification of betalains (betacyanins and betaxanthins contents) were performed
according to the method of Castellanos-Santiago and Yahia (2008) using a Lambda 25 UV-Vis
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spectrophotometer (Perkin Elmer). The betacyanin (BC) content and betaxanthin (BX) contents were
calculated using the following equation:
󰇛 󰇜 󰇟󰇛󰇜󰇛󰇜󰇛󰇜󰇛󰇜󰇠
󰇛󰇜󰇛󰇜󰇛󰇜
Ec. (4)
where: A is the absorbance value at the absorption maxima of 535 and 483 nm for betacyanins and
betaxanthins, respectively, DF is the dilution factor, Vd is the volume of the dry pulp solution (ml),
Wd is the weight of the dry pulp (g), and L is the path length (1 cm) of the tray. The molecular weights
(Mw) and molar extinction coefficients (ε) of betanin [Mw=550 g/mol; ε= 60,000 L/(mol cm) in water
at 536 nm] and indicaxanthin I [Mw= 308g/mol; ε= 48,000L/(mol cm) in water at 481 nm] were used
for the quantification of betacyanins and betaxanthins. Measurements were performed in triplicate
and the mean values with standard deviation were reported.
Individual betacyanins were analyzed by high pressure liquid chromatography using the method
described by Castellanos and Yahia (2008). Briefly, samples were filtered through 0.45 μm nylon
filters, 20 μL were injected into a Thermo Scientific Dionex Ultimate 3000 UHPLC equipped with a
Thermo Scientific reversed-phase column (C18, 250 x 4.6 mm; drop size 0.5 μm), and a UV detector
(DAD Varian ProStar model 410, Palo Alto, CA). The mobile phase A: methanol/KH2PO4 0.05 M (18:82
v/v) pH=2.75 with phosphoric acid, and the mobile phase B: methanol, were adjusted to a gradient of
100 % A to 80 % A and 20 % B in 20 min at a flow rate of 1 mL/min. Detection was performed at 536
nm for betacyanins.
Antioxidant Activity (AA): The AA was determined according to the spectrophotometric method
according to Kuskoski et al. (2005), based on the measurement of DPPH• radical absorbance. First, 0.1
mL of extract was added to 3.9 mL of a DPPH• radical solution (100 µM), vortexed (Vortex-1, Genie;
Scientific Industries Inc.), and stored in the dark for 3 h. The absorbance was then measured at 517
nm using a Lambda 25 UV-Vis spectrophotometer (Perkin Elmer). A Trolox (0.08-1.28 mM)
calibration curve was used. For juice the method described by Ruiz-Gutiérrez et al. (2014) was used.
For encapsulated powders, the extracts were obtained according to the method described by Pitalua
et al. (2010) with modifications; encapsulated powder (2.5 g) was dispersed in 50 mL of methanol and
deionized water (1:1 v/v). The dispersions were homogenized for 15 s using an Ultra-Turrax (basic
IKA T18) with a dispersing tool (S18N19G; IKA Works Inc.). The mixture was centrifuged at 3000 xg
for 10 min (Centra centrifuge CL3R IEC; Thermo Electron Corporation). To ensure complete
extraction, the process was repeated using 25 mL of methanol and deionized water (1:1 v/v). The
supernatants were collected, filtered through a 0.22 µm pore polyethylene filter (Millipore Corp.,
Bedford, MA, USA). Results were expressed as µmol Trolox equivalents per gram of sample on a dry
basis (mg TE/g). The determinations were performed in triplicate and the mean values were
expressed as the standard deviation.
2.3 Experimental design and statistical analyses
A 2×2 completely randomized factorial design in duplicate was applied. The independent
variables were carrier agent (25 and 75 %) and the inlet drying temperature (150 and 180 ºC). The
dependent variables were the physical and chemical properties of the encapsulated powder. Data
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analysis was performed using Minitab (2018) software. The effect of the factors was determined using
a two-way analysis of variance (ANOVA), and the Comparisons of means were calculated using
Tukey's test at a significance level of 95 %.
3. Results y discussions
3.1 Beetroot stem juice characterization
Table 1 shows the characteristics of beetroot stem juice, the juice presented a pH of 5.98, a
low soluble solids content (3.1 ºBrix). The color presented a low brightness and tendencies to redness
and blueness due to the presence of betalains and chlorophyll, respectively. The chemical
characteristics showed a total betalains content of 107.85 mg/kg, divided in 71.78 for betacyanins and
36.07 mg/kg of betaxanthins, showing that the main betalains group is betacyanins (red-purple
color), and an antioxidant activity of 16.52 ± 0.74 (mmol TE/100g). Beetroot stem juice presents similar
characteristics of pH, color and total betalains content to beetroot juice with values of pH = 6.5, 5.62,
2.13, -1.26 for L, a and b and 135.75 mg betalains/kg beetroot, according to Pitalua et al. (2010), the
main difference is in soluble solids which is almost 4 times lower than beetroot juice.
Table 1. Characteristics* of beetroot steam juice and A. vera gel
Tabla 1. Características* de jugo de tallo de betabel y gel de A. vera
Determination
Juice
A. vera
pH
Soluble solids
Luminosity
Green-red tendency
Blue-yellow tendency
Antioxidant activity (mmol TE/100g)
Betacyanins (mg/kg)
Betaxanthins (mg/kg)
5.98 ± 0.1
3.1 ± 0.1
23.8 ± 0.02
2.75 ± 0.03
-2.37 ± 0.03
16.52 ± 0.74
71.78 ± 1.88
36.06 ± 1.24
4.66 ± 0.02
0.87 ± 0.02
36.15 ± 0.1
0.54 ± 0.01
2.05 ± 0.05
6.71± 0.22
-
-
For A. vera gel the pH was 4.66 and soluble solids of 0.87 ºBrix, results according to IASC
(International Aloe Science Council), low soluble solids and acid pH. The color presented a slight
tendency to yellow and a value of 6.72± 0.23 (mmol TE/100g) for antioxidant activity.
3.3 Encapsulated powders of beetroot stem juice
Four different powders were obtained and are showed in the Fig. 1, corresponding to the
combinations of low content of A. vera in the mixture 25A:75B and high content of A. vera in the
mixture 75A:25B at low spray drying temperature (150 ºC) and high spray drying temperature (180
ºC).
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a)
b)
c)
d)
Figure 1. Encapsulated powder of beetroot stem using different content of A. vera as carrier and dried at
different temperatures: % A = A. vera gel and % B = beet stem juice; a) 25A:75B, 150 ºC; b) 25A:75B, 180 ºC; c)
75A:25B, 150 ºC; d) 75A:25B, 180 ºC.
Figura 1. polvo encapsulado de tallo de betabel secado a diferentes temperaturas utilizando A. vera como
acarreador: % A = Gel de A. vera y % B = jugo de tallo de betabel; a) 25A:75B, 150 ºC; b) 25A:75B, 180 ºC; c)
75A:25B, 150 ºC; d) 75A:25B, 180 ºC.
In general, the powders obtained with low A. vera (25 %) show a more red-purple color, due to the
higher content of betalains in the mixture, but the effect of the drying temperature is visible. The
powder dried at higher temperature (180 ºC) showed a red-purple color more intense than the
powder dried at lower temperature (150 ºC). On the other hand, the color in the powders obtained
with high content of A. vera in the mixture (75 %) at both drying temperatures is very similar the
color, then, it can be related to the reduction of the effect of drying temperature.
The analysis of the stability characteristics of the encapsulated powder of beetroot steam using A.
vera as a carrier is presented in Table 2. The powders of the four treatments had a low aw value,
around to 0.1, this value ensures the stability of the powder and no differences were found between
treatments. Some studies reported a low aw values for powders obtained by spray drying as Ruiz-
Gutiérrez et al. (2014) reported aw values in red cactus pear encapsulated with soluble fiber as carrier
(0.097 - 0.138) and Muñoz (2014) in beetroot juice dried using maltodextrin and grenetin as carriers
(0.019 - 0.029).
Table 2. Characteristics* of encapsulated powder of beetroot steam by spray drying using A. vera
as carrier
Tabla 2. Características* de polvo encapsulado de tallo de betabel secado por aspersión utilizando
A. vera como acarreador
Temperature (ºC)
Mixture (A-B)
Moisture (%)
aw
pH
150ºC
25:75
9.786 ± 0.038a
0.127 ± 0.001a
5.86 ± 0.07a
180ºC
25:75
9.279 ± 0.419a
0.118 ± 0.012a
5.88 ± 0.02a
150ºC
75:25
7.154 ± 0.101b
0.130 ± 0.001a
5.37 ± 0.03b
180ºC
75:25
7.079 ± 0.026b
0.120 ± 0.007a
5.47 ± 0.05b
*Mean ± standard deviation. A (%) = A. vera; B (%) = beetroot steam juice. Means with different
letter by characteristic indicate significative (p<0.05) difference, by Tukey’s test.
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The moisture contents are low, values ranging from 7.079 ± 0.026 to 9.786 ± 0.038 and the pH values
ranged from 5.37 ± 0.03 to 6.88 ± 0.02, for both variables the significant effect (p<0.05) of the A. vera
content as a carrier in the mixture is observed. The encapsulated powders with 25 % of A. vera present
higher moisture and pH values than the encapsulated powders with 75 % of A. vera, because A. vera
is a gel and by adding more in the mixture there is a high overall moisture content that is dried under
the same temperature condition. Moisture contents are similar to curcumin powders (1.9 % to 10.1
%), slightly higher than whey protein isolate powders for blackcurrant concentrate and higher than
maltodextrin and trehalose matrices used for roselle anthocyanins, reported by Guo et al. (2020), Wu
et al. (2021) and Millinia et al. (2024). Moisture values tend to decrease with increasing amount of
carrier used. The pH values are suitable for the stability of the pigments, betalains are generally
stable in the pH range of 4–6 (Martins et al., 2017), the pH values are statistically (p<0.05) lowest in
powders with a 75 % A. vera content (75 %) due to the low pH of the gel.
Other important properties in the encapsulated powders are bulk density, glass transition
temperature and water solubility index, these properties are important in relation to the behavior of
the encapsulated powder during transportation, storage and application and are presented in Table
3.
Table 3. Properties* of encapsulated powder of beetroot steam by spray drying using A. vera as carrier
Tabla 3. Propiedades* de polvo encapsulado de tallo de betabel secado por aspersión utilizando A. vera como
acarreador
Temperature (ºC)
Mixture (A-B)
Bulk density (g/mL)
Tg (ºC)
WSI
150ºC
25:75
0.730 ± 0.028ab
55.98 ± 2.81a
89.49 ± 0.23a
180ºC
25:75
0.755 ± 0.007a
53.80 ± 4.22a
86.60 ± 0.39b
150ºC
75:25
0.680 ± 0.000b
48.36 ± 2.90a
81.84 ± 1.30c
180ºC
75:25
0.670 ± 0.014b
51.60 ± 0.91a
86.89 ± 0.08ab
*Mean ± standard deviation. A (%) = A. vera; B (%) = beetroot steam juice; Tg = Glass transition temperature;
WSI = Water solubility index. Means with different letter by properties indicate significative (p<0.05)
difference, by Tukey’s test.
The bulk density ranges from 0.670 ± 0.014 to 0.755 ± 0.007 g/mL, although the values of the different
treatments are close, there is a significant (p<0.05) effect of both the amount of A. vera used and the
interaction of this factor with temperature, being the effect of the amount of A. vera being more
important. The encapsulated powders produced with 75 % of A. vera have a lower density than the
encapsulated powders produced with 25 % of A. vera, because the solids content in the A. vera gel is
lower than the solids content in the beetroot stem juice. Bulk densities slightly lower than the present
study have been reported, 0.497–0.525 g/mL for Amaranthus extract (Tabio-García et al., 2022) using
Opuntia ficus-indica mucilage and maltodextrin as a carrier.
The Tg was statistically (p>0.05) equal among the four encapsulated powders, values ranging from
48.36 ± 2.90 to 55.98 ± 2.81 ºC. This temperature indicates the temperature at which the product
changes its density, hardness, and stiffness and then retains its properties until reaching a
temperature of about 50 ºC is reached. This Tg value is better than that reported by Ruiz-Gutiérrez et
al. (2014) for encapsulated powders prepared by spray drying but using soluble fiber as a carrier,
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who reported values from 32.27 to 37.69 ºC, and similar to the Tg (43.74-49.65 ºC) reported for
powders dried at 150 ºC using maltodextrin and Opuntia ficus-indica mucilage as a carrier (Tabio-
García et al., 2022).
The water solubility index (WSI) was significantly (p<0.05) affected by the amount of A. vera used in
the mixture (0.002) and the interaction of drying temperature - amount of A. vera (0.001), the values
ranged from 81.84 ± 1.30 to 89.49 ± 0.23, respectively, these values were high compared to the WSI
reported by Ruiz-Gutiérrez et al. (2014), who reported WSI values of 0.64 to 0.764 for red cactus pear
juice dried with soluble fiber, and similar to Tabio-García et al. (2022), who reported 82.96–92.81 %
using maltodextrin and Opuntia ficus-indica mucilage as a carrier for encapsulation of Amaranthus
extract. Although the WSI values are similar among the powders, the encapsulated powder with 75
% of A. vera dried at 150 ºC presented the lowest WSI, and the encapsulated powder with 25 % of A.
vera dried at 180 ºC presented the highest WSI.
The analysis of the morphology of the powders, obtained by scanning electron microscopy (SEM) is
shown in Fig. 2, the images showed a matrix in the form of spherical capsules with little
agglomeration. Fig. 2a and 2b correspond to encapsulated powders obtained with 25 % of A. vera
and show more spherical and less compressed capsules compared to encapsulated powders
obtained with 75 % of A. vera (Fig. 2c and 2d). This compression characteristic is normal in spray
drying as it can occur at the time of elimination of moisture. The collapse of the wall is more
noticeable in Fig. 2c and 2d, where A. vera predominates in the mixture, since it contains a higher
moisture content than the beetroot stem juice, causing a more compressed sphere. Therefore, the
removal of water deforms the spherical particle in the case of the two powders. Different matrices
produce different coating wall densities, and consequently, different shapes and textures, the
maltodextrin-trehalose matrix, as a carrier, achieved more spherical shapes and smoother surfaces
than in the present study, due to the presence of trehalose sugar (Millian et al., 2024) or drying at a
lower inlet temperature (120 ºC). Powders obtained at similar drying temperatures show similar
shapes in the capsules of the powders obtained with a 25 % of A. vera of the present study (Ruiz-
Gutiérrez et al. 2014).
Figure 2. Encapsulated powder of beetroot stem with different content of A. vera as a carrier and dried at
different temperatures; a) 25 % A. vera at 150 ºC, b) 25 % A. vera at 180 ºC, c) 75 % A. vera at 150 ºC, d) 75 % A.
vera at 180 ºC.
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Castillo-Altamirano et.al
Figura 2. Polvo encapsulado de tallo de betabel secado a diferentes temperaturas utilizando A. vera como
acarreador; a) 25 % A. vera a 150 ºC, b) 25 % A. vera a 180 ºC, c) 75 % A. vera a 150 ºC, d) 75 % A. vera a 180 ºC.
However, in this case, the temperature does not affect either of the two powders, as these are very
high temperatures that are capable of eliminating most of the water in the encapsulates.
The color parameters (L*, a* and b*) are presented in Table 4. The luminosity (L* parameter) presents
values ranging from 26.74 to 53.26, L* was significantly (p<0.05) affected by percent of A. vera used
in the mixture, finding that the encapsulated powder produced with 25 % of A. vera tends to dark
color while that the encapsulated powder with 75 % of A. vera tends to white. Powders of whey
protein isolate and black currant concentrate by spray drying showed a similar L* parameter value,
52.81 ± 0.50, (Wu et al., 2021) to powders with 75 % A. vera content.
Table 4. Color parameters* of encapsulated powder of beetroot steam by spray drying using A.
vera as carrier dried at different temperatures.
Tabla 4. Parámetros de color de polvo encapsulado de tallo de betabel secado por aspersión
utilizando A. vera como acarreador.
Temperature (ºC)
Mixture (A-B)
L*
a*
b*
150
25:75
26.74 ± 0.08b
15.91 ± 0.85b
-1.00 ± 0.03b
180
25:75
32.99 ± 5.62b
22.43 ± 1.08a
-1.54 ± 0.02c
150
75:25
53.26 ± 0.42a
8.44 ± 0.01c
13.88 ± 0.07a
180
75:25
51.32 ± 0.68a
8.07 ± 0.01c
13.96 ± 0.12a
*Mean ± standard deviation. A (%) = A. vera; B (%) = beetroot steam juice. Means with different
letter by color parameter indicate significative (p<0.05) difference, by Tukey’s test.
For the a* parameter, the significant effects were due to the A. vera used in the mixture and the drying
temperature, showing more tendency to red color the encapsulated powder prepared with 25 % of
A. vera, values were 15.91 ± 0.85 and 22.43 ± 1.08 at 150 and 180 ºC of drying temperature,
respectively. For the encapsulated powder with 75 % of A. vera the values of parameter a*, for both
temperatures, were around 8, being significantly (p<0.05) lower than the encapsulated powder
elaborated with 25 % of A. vera. The difference is that this powder has less beetroot stem juice in the
mixture, then less betalains, reducing the red tendency. For the b* parameter the effects and
differences are the same as for the a* parameter, but the high b* values (around 13) were for
encapsulated powders with 75 % of A. vera, indicating a tendency to yellow. The low b* values were
for encapsulated powders with 25 % of A. vera, indicating a tendency to blue, and the powder dried
at 180 ºC presents the tendency to blue (-1.54 ± 0.02). At high drying temperatures, there may be
greater degradation of betaxanthins (yellow-orange pigments), reducing the tendency to +b*
(yellow).
The betalains analyses showed the expected trend, the betalains content is shown in Fig. 3. The
encapsulated powders with 25 % A. vera had a higher betalains content, because the mixture
contained more beetroot stem juice. No significant differences (p>0.05) were observed by drying
temperature. However, contrary to other studies, in which an increase in spray drying temperature
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was found to lead to a decrease in betacyanin content, however, the temperatures at which this effect
was observed were > 180 ºC (Ruiz-Gutiérrez et al., 2014).
Figure 3. Betalains content on encapsulated powder of beetroot stem using different contents of A. vera as carrier
and dried at different temperatures. ■ Betacyanins and ■ Betaxanthins. Mean ± standard deviation. A = A. vera;
B = beetroot steam juice. Means with different letter by betalain group indicate significant (p<0.05) difference,
by Tukey's test.
Figura 3. Contenido de betalainas de polvo encapsulado de tallo de betabel utilizando diferentes contenidos de
A. vera como acarreador a diferentes temperaturas de secado. ■ Betacianinas y ■ Betaxantinas. Media con
diferente letra por grupo de betalaína indican diferencia significativa (p<0.05), mediante prueba de Tukey.
Highlighting that in the encapsulated powders obtained with 25% A. vera in the mixture, the
betacyanin content is about double compared to betaxanthins, being visually noticeable (Figs. 1a and
1b), presenting contents ranging from 1.185 to 1.343 mg/g and from 0.671 to 0.705 mg/g, for
betacyanins and betaxanthins, respectively. However, encapsulated powders obtained with 75 % A.
vera, the contents of betacyanins and betaxanthins do not present differences (p>0.05) showing
contents around 0.35 mg/g of total betalains. These powders show a low tendency to red color and
high tendency to yellow color (Table 3), and visually they have a pale beige color (Figs. 1c and 1d).
The total betalains content is similar to that reported by Ruiz-Gutiérrez et al. (2014) for powder of
red cactus pear powder by spray-drying using soluble fiber as a carrier, with values around of 1.4 –
1.8 mg/g for betacyanins and around 0.65 mg/g for betaxanthins were reported.
The analysis of betalains by HPLC helps to identify the individual betalains, especially the betalain
that represents the group (betacyanins or betaxanthins) and which depends on the source, using the
information reported by Slatnar et al. (2015) and Mikołajczyk-Bator & Pawlak (2016), such as
retention times and elution order. Betacyanins are the most important group of pigments or with the
highest content in the beetroot stem as shown by the quantification of total betalains carried out by
UV-Vis spectroscopy (Fig. 3). These are the red-purple pigments and are quantified at a length of
about 536 nm. The main individual pigments in this group are betanin, isobetanin, and neobetanin,
although many more betacyanins may be present, and when these are analyzed by chromatography
the order in which they appear in the chromatogram is betanin, isobetanin and neobetanin (Slatnar
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Castillo-Altamirano et.al
et al., 2015). The analyses for changes in betacyanins are summarized in Table 5, where the areas of
the four peaks found in the chromatograms are reported.
Table 5. Individual betacyanins of encapsulated of beetroot stem powder using different levels of A. vera as
carrier and different spray drying temperatures.
Tabla 5. Betacianinas individuales de polvo encapsulado de tallo de betabel utilizando diferentes contenidos
de A. vera como acarreador a diferentes temperaturas de secado.
Temperature (ºC)
Mixture (A-B)
Peak 1
(mAU)
Peak 2
(mAU)
Peak 3
(mAU)
Peak 4
(mAU)
150
25:75
3.50 ± 0.08a
10.93 ± 0.62a
1.44 ± 0.10a
0.54 ± 0.02a
180
25:75
2.53 ± 0.23b
6.10 ± 0.11b
1.03 ± 0.05b
0.35 ± 0.01b
150
75:25
nd
nd
nd
nd
180
75:25
nd
nd
nd
nd
Area mean ± standard deviation. A (%) = A. vera; B (%) = beetroot steam juice; nd = not detected. Means with
different letter by peak indicate significative (p<0.05) difference, by Tukey’s test.
In the analysis of betacyanins in beetroot juice, betanin has been reported as the first peak in
chromatograms, being the peak with more area. However, when beetroot juice is treated thermally,
the first peak has a smaller area than betanin, being a product of betanin degradation, and then
betanin appears as the second peak (Mikołajczyk-Bator & Pawlak, 2016). Then in Table 5, peak 2
could be betanin and the peak 1 is a product of its degradation; both peaks (44.19 and 27.7 %,
respectively) decreased significantly (p<0.05) due to an increase in drying temperature for
encapsulated powder obtained with 25 % of A. vera. Peaks 3 and 4, may be related to some modified
structure of betanin, isobetanin or neobetanin. They are smaller areas than betanin (peak 2) and peak
1, but these were also affected by the increase in drying temperature with reductions of 26.1 and 33.9
%, respectively. Tabio-García et al. (2022) reported that betacyanin losses occur in Amaranthus at 150
ºC during spray drying.
On the other hand, no peaks were found in the encapsulated powders obtained with 75 % of A. vera
in the mixture. This is due to the fact that the content of beetroot stem juice is low and consequently
the content of betacyanins is also low, and in addition the drying temperature causes decreases or
changes in these pigments. While that, betaxanthins (yellow-orange pigments) are the minority
group, composed of vulgaxanthin I and II, as well as miraxanthin V. However, due to their low
content, no peak could be detected during the analysis, for which a wavelength of 481 nm was used.
It has been reported that betaxanthins are more affected by the effect of temperature than
betacyanins (Ruiz-Gutiérrez et al., 2014; Mikołajczyk-Bator & Pawlak, 2016).
Regarding the antioxidant activity of the encapsulated powders, the results are shown in Table 6,
this property had a similar behavior to betalains. The differences were related to the A. vera used in
the mixture. The highest antioxidant activity values were around 1.3 mmol TE/100 g for encapsulated
powders with 25 % of A. vera, for both drying temperatures, being statistically different from
powders with 75 % A. vera dried at 150 ºC with values of 0.18 mmol TE/100 g. The values of
antioxidant activity of encapsulated powders are lower than the antioxidant activity of beetroot stem
juice and A. vera gel (Table 1), due to degradation or transformation of compounds as betalains and
others as polyphenols (Tabio-García et al., 2022).
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Table 6. Antioxidant activity of encapsulated beetroot stem powder with different content of A.
vera as carrier and different spray drying temperature.
Tabla 6. Actividad antioxidante de polvo encapsulado de tallo de betabel utilizando diferentes
contenidos de A. vera como acarreador a diferentes temperaturas de secado.
Temperature (ºC)
Mixture (A-B)
Antioxidant activity (mmol TE/100 g)
150
25:75
1.33 ± 0.13a
180
25:75
1.36 ± 0.03a
150
75:25
0.18 ± 0.05b
180
75:25
nd
Area mean ± standard deviation. A (%) = A. vera; B (%) = beetroot steam juice; nd = not detected.
Means with different letter by peak indicate significative (p<0.05) difference, by Tukey’s test.
4. Conclusion
The use of A. vera gel as a carrier in spray drying led to the obtaining of encapsulated beetroot
stem juice powders. The results show that with increasing A. vera decreased H, pH, BD, Tg, WSI, BC,
BX and the tendency to red, while luminosity and tendency to yellow increased, presenting more
compressed capsules in the powders. Temperature affected the WSI, the color parameters a* and b*
and the individual betacyanins, reducing the individual betacyanins and consequently the tendency
to red. The encapsulated powders obtained showed characteristics that allow the stability of the
powder components, such as low aw (<0.13) and low moisture content (<10 %), pH suitable for the
stability of betalains (5.3-5.88) and Tg suitable for storage (around 50 ºC). Highlighting that the
powder obtained with 25 % A. vera and at a drying temperature of 180 ºC presented the best
characteristics to be used as an ingredient in food formulation, with a water solubility index of 86.6,
a color with a tendency to red-purple and with a content of ≈1.3 mg/g and ≈0.68 mg/g of betacyanins
and betaxanthins, respectively, representing an alternative as a natural pigment to give or improve
the color in food products.
Aknowlendgments
Studies carried out at the Autonomous University of Chihuahua (UACH).
Interes conflict
The authors declare no conflict of interest.
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Castillo-Altamirano et.al
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