Food Chemistry 307 (2020) 125535

Contents lists available at ScienceDirect

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

Microencapsulation by spray-drying of polyphenols extracted from red T

chicory and red cabbage: Effects on stability and color properties
⁎
Francesca Zanonia,b, Martina Primiterraa, Nicola Angelic, Gianni Zoccatellia,b,
a
University of Verona, Department of Biotechnology, Verona, Italy
b
Sphera Encapsulation Srl, Verona, Italy
c
MUSE, Limnology and Phycology Section, Trento, Italy

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

Keywords: The research of antioxidants and natural pigments to replace synthetic molecules is increasingly considering
Red chicory (Chicorium intibus L.) wastes from plant food supply chains. Red chicory (RCH) and red cabbage (RCA) are rich sources of polyphenols
Red cabbage (Brassica oleracea L. var capitata f. (PP), especially anthocyanins, well know natural pigments possessing strong antioxidant capacity and beneficial
rubra) health effects. The aim of this work was to compare different solvents for PP extraction and to evaluate the effect
Polyphenols
of spray-drying encapsulation using modified starch on PP, antioxidant capacity (AOC) and color properties.
Microencapsulation
Antioxidant capacity (AOC)
Methanol:water (70:30) showed the best extraction capacity, while ethanol:water (70:30) extracts displayed
Spray-drying the highest thermal stability. Ethanol:water extracts were spray-dried with a yield of 95–99% for both crops,
while the efficiency of PP encapsulation was 79% (RCA) and 88% (RCH). Encapsulation improved retention of
PP and AOC upon thermal treatment (RCH: 20–30%, RCA: 44–55%) without altering color properties. This
process can be employed for the development of functional foods and supplements.

1. Introduction Muhvic, & Giacometti, 2015).

The research for natural antioxidants and the growing importance
in the management and requalification of industrial food wastes is in-
creasingly considering plant food by-products as source of valuable
molecules, e.g. polyphenols, carotenoids and fibers (Lante, Nardi,
Zocca, Giacomini, & Corich, 2011). The exploitation of this type of
resources gives the possibility to lower the amount of food wastes,
hence diminishing the negative impact on the environment due to the
phytotoxic effect of the high organic matter content (Lavelli, Harsha,
Laureati, & Pagliarini, 2017).
In particular, among the different classes of molecules with health
promoting effects, during the last 10 years, researchers and food in-
dustries have been focusing their attention on polyphenols. The reasons
rely on the fact that they are present in great abundance in our diet, and
are involved in the prevention of various chronic-degenerative
pathologies such as cardiovascular diseases (Scalbert, Manach, Morand,
Remesy, & Jimenez, 2005), diabetes (Matsui, Ogunwande, Abesundara,
& Matsumoto, 2006), neurodegenerative diseases (Levites, Amit,
Youdim, & Mandel, 2002) and osteoporosis (Dudaric, Fuzinac-Smojver,
Red chicory (Chicorium intibus L.) and red cabbage (Brassica oleracea
L. var capitata f. rubra) are both important sources of polyphenols.
Moreover, the wastes produced during their processing, consisting
mainly of leaves and steams, can reach up to 40–50% of the total
harvested material, that for the great part is still nutritionally and hy-
gienically valid especially in the case of the fourth-range production
where edible material is eliminated also to improve the aspect of the
food and increase the acceptability of the consumer; Lante et al., 2011;
Llorach, Tomas-Barberan, & Ferreres, 2004).
RCH is a typical winter vegetable indigenous to Europe, North and
Western America that has gained attention for its content of phyto-
chemicals with potential health benefits, such as phenolic acids, fla-
vonoids and anthocyanins (Bais & Ravishankar, 2001). The high con-
tent of anthocyanins, that account for the 50–55% of the total phenolic
compounds, is responsible for the characteristic deep red color of the
leaves. Among all the leafy vegetables that are consumed raw, red
chicory has been recognized to have the highest content of polyphenols
(Gazzani, Daglia, Papetti, & Gregotti, 2000).
RCA, in the past indigenous of the Mediterranean zone, is now one

Abbreviations: RCH, red chicory; RCHE, red chicory extract; RCA, red cabbage; RCAE, red cabbage extract; EtOH, ethanol, MetOH, methanol; aw, water activity; GA,
gallic acid; GAE, gallic acid equivalents; TE, Trolox equivalents; PP, Polyphenols; TP, total polyphenols; SP, superficial polyphenols; EE, Encapsulation efficiency; YE,
yield of encapsulation; Abs, absorbance; f.w., fresh weight; OSA, octenyl succinic anhydride; ABTS, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid
⁎
Corresponding author at: Dipartimento di Biotecnologie, Università degli Studi di Verona, Strada Le Grazie, 15 – CV1, I-37134 Verona, Italy.
E-mail address: gianni.zoccatelli@univr.it (G. Zoccatelli).

https://doi.org/10.1016/j.foodchem.2019.125535
Received 14 May 2019; Received in revised form 5 September 2019; Accepted 13 September 2019
Available online 30 September 2019
0308-8146/ © 2019 Elsevier Ltd. All rights reserved.
F. Zanoni, et al.
of the most important vegetables grown around the world (Wiczkowski,
Szawara-Nowak, & Topolska, 2013). The abundance and the relative
low cost of production, if compared to other vegetables rich in poly-
phenols (e.g. black carrot and purple potatoes), make it an attractive
source of natural red pigments (Bridle & Timberlake, 1997). Its use has
been reported in the treatment of inflammatory bowel disease
(Zielińska et al., 2015), and in the attenuation of cardiac and hepatic
oxidative stress (Sankhari, Thounaojam, Jadeja, Devkar, &
Ramachandran, 2012).
It is worth considering that the production and harvest of these
vegetables are seasonal activities and the by-products obtained from
their processing need to be handled in a limited time frame, since once
extracted, polyphenols are highly susceptible to degradation (Patras,
Brunton, O'Donnell, & Tiwari, 2010).
Spray-drying has been widely used to dry a number of foods and
pharmaceutical preparations, among which many containing heat-
sensitive molecules such as polyphenols (Shishir & Chen, 2017). It is the
most common technique used to turn liquids with high solid content
into powders. In addition, it is recognized as an economic process by
which it is possible to produce high quality powders. To our knowledge,
no work about the stabilization of polyphenols from red chicory
through spray-drying has been addressed, and only two works about the
stabilization of polyphenols from red cabbage have been described
(Bernstein & Noreña, 2015; Hongmei & Meng, 2015). In these studies
only the production and characterization of the powders were tackled
without evaluating their performance in a simulating medium.
The aim of this work was to assess the quality and the thermal
stability of phenolic extracts from red chicory and cabbage leaves ob-
tained by different solvents (water, and hydro-alcoholic mixtures of
ethanol and methanol) and to improve the stability of polyphenols and
antioxidant capacity by spray-drying encapsulation without altering the
color properties of the pigments.
2. Materials and methods:
2.1. Materials
Fresh Cichorium intybus L. (red chicory var. Chioggia) and Brassica
oleracea L. (red cabbage var. Capitata f. rubra) were purchased from a
local market. Modified starch CAPSUL® was provided by Ingredion
(Illinois, US). Folin-Ciocalteau reagent, sodium carbonate, formic acid,
methanol, ABTS, potassium persulfate, 37% chloridric acid, ethyl
acetate, Trolox®, gallic acid and ethanol were obtained from Sigma
Aldrich (St. Louis, MO, US).
2.2. Extraction of polyphenols
Polyphenols were extracted from RCH and RCA leaves combining
different protocols with some modifications (Cefola, Carbone, Minasi, &
Pace, 2016; Heimler, Isolani, Vignolini, & Romani, 2009; Innocenti
et al., 2005; McDougall, Fyffe, Dobson, & Stewart, 2007; Pereira, de
Arruda, & Stefani, 2015). Briefly, 500 g of RCH fresh leaves were
ground with a SAMA K45 cutter (Dito SAMA, Italy). The obtained
sample was extracted with the solvent for 4 h in continuous agitation
(ratio sample: solvent 1:1). Formic acid was used to adjust the pH of the
sample to 3.3–3.5. The extract was filtered through Whatman filter
paper in a Buchner funnel. The obtained solution was kept at 4 °C in the
dark until use. Three different extraction solvents were tested: water,
30% water:70% EtOH and 30% water:70% MetOH. The same opera-
tions were conducted with RCA. Methanol and ethanol were then re-
moved by evaporation with a rotary evaporator (Büchi, Switzerland)
prior use/storage.
2.3. Polyphenols quantification
The Folin-Ciocalteau assay was performed following a method
2
Food Chemistry 307 (2020) 125535
previously proposed (Singleton, Orthofer, & Lamuela-Raventos, 1999)
with some modifications. Briefly, the reaction was conducted in a 96
well micro-plate (Sarstedt, Germany). An amount of 5 μl of each sample
was incubated for 3 min with 150 μl of Folin-Ciocalteau reagent (pre-
viously diluted at 1:15 with water). Finally, 40 μl of 20% sodium car-
bonate solution was added and left to react for 30 min in the dark. The
absorbance was measured using a micro-plate reader (BioTek, Milan,
Italy) at 750 nm. Water was used as blank. The calibration curve was
carried out using GA as standard. The results were expressed as milli-
grams GAE per gram of f.w. All the experiments were performed in
triplicate.
2.4. Antioxidant capacity
The procedure proposed by Thaipong and co-workers was followed
(Thaipong, Boonprakob, Crosby, Cisneros-Zevallos, & Byrne, 2006)
with some modifications. The stock solutions consisting of 7.4 mM
ABTS and 2.6 mM potassium persulfate were prepared. The working
solution was prepared by mixing the two stock solutions in equal vo-
lumes and allowing them to react for 12 h at room temperature in the
dark. The solution was then diluted opportunely with phosphate-buf-
fered saline (10 mM phosphate buffer, 150 mM NaCl, pH 7.4) to obtain
an absorbance of 0.75 units at 734 nm. Fresh ABTS solution was pre-
pared for each assay. Samples (20 μl) were placed in a 96-well plate and
left to react with 180 μl of the ABTS solution for 2 h in the dark. The
absorbance was measured at 734 nm using a micro-plate reader. Trolox
served as standard. The results were expressed in μmols TE per gram
f.w. All the experiments were performed in triplicate.
2.5. Thermal stability
The stability of RCHE and RCAE before and after encapsulation was
studied at 100 °C. The analyses were conducted placing samples of the
extracts or of the powders (100 mg resuspended in 1 mL of water, final
pH 3.5–3.6) in a thermostatic water bath for 3 h. Samples were col-
lected every hour to quantify PP and AOC. In the case of powders 5 ml
of ethanol was added to the samples that were centrifuged at 4500g for
10 min to remove insoluble material. PP and AOC were measured on
the supernatants.
2.6. Microparticles production and characterization
2.6.1. Microencapsulation by spray-drying
Among the different extracts those carried out in ethanol were
chosen to be microencapsulated. CAPSUL® modified starch was rehy-
drated directly in 100 ml of the extracts for 1 h prior its use at a final
concentration of 15%. The atomization process was performed with the
use of a Mini-Spray dryer B-290 (Büchi). The conditions used were as
follows: drying air flow rate 40 m3/h; inlet air temperature 140 ± 3;
outlet air temperature 70 ± 3 and a feed flow rate of 5 mL/min. The
formed microparticles were collected at the bottom of the cyclone se-
parator. The RCHE and RCAE powders were kept in aluminum sealed
bags and stored at −20 °C until use.
2.6.2. Yield of encapsulation
The YE was calculated as percentage of the ratio between the
powder collected at the end of the process and the amount of solid used
to initially feed the spray-drying system. The YE was expressed as:
g of collected powder
YE% = × 100
g of total initial solid
2.6.3. Water activity
The Aw of the spray-dry powders was measured in triplicate using a
Rototronic (Bassersdorf, Switzerland) device filling the 4.5 cm diameter
F. Zanoni, et al.
disposable plates with the powder and incubating at 20 °C.
2.6.4. Encapsulation efficiency
TP and SP of the powders were evaluated following a protocol
previously proposed (Mandavi, Jafari, Assadpour, & Ghorbani, 2016)
with some modifications.
In order to obtain the SP, 100 mg of powder was incubated with
10 ml of ethanol. The solution was left in agitation for 1 min in a rotator
mixer, and the sample was centrifuged at 4500g for 10 min. The su-
pernatant was recovered and filtered at 0.45 μm.
For the evaluation of the TP, 100 mg of powder was resuspended in
1 ml of water. The sample was placed in a sonication bath for 30 min at
room temperature to allow the rupture of the microparticles in solution.
Ethanol (10 ml) was added and the solution was left in agitation for
30 min in a rotator mixer. The sample was centrifuged at 4500g for
10 min and then filtered at 0.45 μm. The quantification of the poly-
phenols content was performed by Folin-Ciocalteau as described above.
The EE was calculated using the subsequent formula:
TP − SP
EE% = × 100
theoretical PP content
2.6.5. Optical microscopy
Spray-dried powders were resuspended in mineral oil. The oil was
dispersed on a glass slide and images were taken using an EVOS optical
microscope (Thermo Fisher Scientific, US). Microparticles diameters
have been measured by analyzing digital images using the software
ImageJ (http://imagej.nih.gov/ij). One hundred microparticles have
been measured for each batch.
2.6.6. Scanning electron microscopy
The observation of microparticles surface morphology was carried
out using a scanning electron microscopeZeiss XVP (Carl Zeiss SMT
Ltd., Cambridge, UK). The samples were attached to a graphite tape and
then covered with a thin film of gold at high vacuum (Agar Sputter
Coater B7340, Agar Scientific Ltd., Stansted, Essex, UK).
2.7. UV–vis spectra analysis
UV–vis spectra of RCHE and RCAE before and after encapsulation
were analyzed using a Evolution 201 UV–vis spectrophotometer
(Thermo Scientific). The samples were diluted 1:40 in 10 different
media consisting of water in which the pH was modified opportunely
with HCl and NaOH to represent a pH range from 1 to 10. The spectra
were recorded between 300 and 750 nm.
2.8. Statistical analysis
All measurements, if not differently stated, were performed in tri-
plicates. The statistically significant differences among the obtained
data were analyzed using t-test. The differences were considered sig-
nificant with p < 0.05. The Pearson coefficient was computed to test
the correlation between polyphenol content of the extracts and anti-
oxidant capacity. Data are reported as the average ± the standard
deviation.
3. Results and discussion
3.1. Polyphenol extraction
Three different solvents were used to extract polyphenols from the
edible parts of RCH and RCA: water, and hydro-alcoholic mixtures of
ethanol and methanol. In general methanol exhibited the best ex-
tracting capacity leading to PP content in RCHE of about 1.70 mg GAE/
g f.w. (Fig. 1 A) not far from the values observed in other works dealing
with different red chicory varieties (D'Evoli et al., 2013; Heimler,
3
Food Chemistry 307 (2020) 125535
Isolani, Vignolini, Tombelli, & Romani, 2007; Innocenti et al., 2005;
Lavelli, 2008).
In the case of RCAE the amount of extracted PP was higher, on
average 2.20 mg GAE/g f.w. (Fig. 1A), confirming previous data that
indicated a higher PP content of RCA leaves in comparison to RCH
(Innocenti et al., 2005; Kaulmann, Jonville, Schneider, Hoffmann, &
Bohn, 2014). A significant difference was found between the RCAE
EtOH and RCAE MetOH, with the latter presenting a PP content about
25% higher.
The AOC investigated by ABTS assay showed a good correlation
with the PP content in the case of RCHEs (R = 0.85) and a strong
correlation (R = 0.99) in the case of RCAEs (Fig. 1 B).
3.2. Stability assessment of the extracts
To evaluate the stability of the extracted PP, RCHE and RCAE were
treated at 100 °C for 3 h to simulate the boiling process and the amount
of retained PP and AOC was measured. Fig. 2 shows the percent re-
tainment of polyphenol and AOC as a function of the time of boiling.
For both the samples a significant loss of PP was observed after 3 h of
treatment. In the case of RCHE, the water extract showed the lower
stability, with a loss of PP of 90% after 3 h (Fig. 2A) and a decrease of
the AOC of 80% (Fig. 2C). The trend displayed by this sample was quite
interesting since within 2 h of treatment it showed the higher PP re-
tainment but after 3 h this value dramatically decreased.
Differently from the water extract, RCHE MetOH and RCHE EtOH
showed higher PP and AOC% retainment (Fig. 2A, C). In particular,
RCHE EtOH displayed 65% retainment of AOC, even though the AOC of
the crude extract was definitely lower than that of RCHE MetOH
(Fig. 1B).
In the case of RCA the time-dependent thermal treatments of the
two hydro-alcoholic extracts showed clearly different patterns, being
PP and AOC% retainment of the RCAE EtOH 25–30% higher than that
of RCAE MetOH (Fig. 2B, D).
In particular, the PP% retainment of RCAE MetOH was very close to
that of RCAE H2O (Fig. 2B) while, in the case of the AOC, the stability of
the MetOH extract was definitely the lowest (Fig. 2D).
It is noteworthy that the PP and AOC% retainment of RCAE EtOH
(42% and 40% respectively, Fig. 2B, D) were lower if compared to the
PP and AOC% retainment of RCHE EtOH (64% and 70% respectively,
Fig. 2A, C) indicating that somehow the polyphenols extracted from red
cabbage are less stable that those extracted from red chicory. This result
was unexpected since anthocyanins from red cabbage, which represent
a consistent amount of polyphenols, when extracted in similar condi-
tions were shown to be very stable due to their acylation (Wiczkowski
et al., 2013). The reason of these apparently contradictory results is
probably the different ratio of anthocyanins to other polyphenols
(mainly hydroxycinnamic acids). Indeed, while in RCH (var. Chioggia)
this value is around 0.20, so with hydroxycinnamic acids being 5 times
higher than anthocyanins (Tardugno, Pozzebon, Beggio, Del Turco, &
Pojana, 2018), in RCA this ratio is close to 4.0, so with anthocyanins
definitely more concentrated than hydroxycinnamic acids (Mizgier
et al., 2016), and being anthocyanins sensibly more labile to degrada-
tion than other polyphenol classes (Brownmiller, Howard, & Prior,
2008), the heating process produced a more marked degradation of
RCA polyphenols despite the protective effect of acylation. Due to the
higher stability of the EtOH extracts, and to the fact that methanol
could raise some safety concerns, RCHE EtOH and RCAE EtOH were
chosen for the subsequent microencapsulation.
3.3. RCHE and RCAE PP encapsulation
For this process CAPSUL® was selected as encapsulant matrix. This
is a native starch modified with OSA. Due to the amphiphilic character
of the grafted functional groups, OSA-starch presents excellent film
forming properties (Sweedman, Tizzotti, Schafer, & Gilbert, 2013).
F. Zanoni, et al.
Fig. 1. PP content (panel A) and AOC (panel B) of RCHE and RCAE extracts. Statistically significant differences (p < 0.05) within each extract are indicated by
different letters.
Fig. 2. Time-dependent percentage retainment of PP (A and B) and AOC (C and D) of RCH (A and C) and RCA (B and D) extracts upon heat treatment at 100 °C for
3 h.
The obtained powders were characterized by a pink-purple color in
the case of RCHE (Fig. 3A) and of an intense violet color in the case of
RCAE powder (Fig. 3B).
Optical microscope observation (Fig. 3C, D) showed the presence of
particles with different dimensions, ranging from 1 μm to 30 μm. The
particles seem to be present as agglomerated structures composed by
encapsulates, visible in both the powders and represented by dark
purple spots, and by small transparent empty particles (similar to in-
clusion bodies) disposed around the encapsulates containing the PP.
Scanning electron microscopy analysis of RCAE powder (Fig. 3F)
confirmed what observed by optical microcopy: indeed, irregular and
spherical particles with different dimensions were visible. In both the
powders (Fig. 3E, F) the strong presence of dented collapsed structures
was observable, with smaller particles that tended to aggregate among
them or with bigger particles. These structures were more visible in the
4
Food Chemistry 307 (2020) 125535
case of RCHE powder. This peculiar morphology could be due to the
low feed rate applied during the drying step that, coupled to the low
amount of solids, renders the drying process too fast leading to the
production of small and collapsed particles (Garcia-Tejeda, Salinas-
Moreno, Hernandez-Martinez, & Martinez-Bustos, 2016; Osorio et al.,
2010). The combination with other materials, i.e. whey proteins, may
improve the surface smoothness and decreased the surface indentation
(Sheu & Rosenberg, 1998). Indeed Bernstein & Noreña (2015) obtained
similar dented particles when using gum arabic as a carrier, while the
same process based on polydestrose gave globular and smooth particles.
The YE was 95.2 ± 2.0% in the case of RCHE and 99.1 ± 0.5% in
the case of RCAE. These data were difficult to compare for the scarce
literature dealing with the encapsulation of these plant extracts. The
water activity of the powders was 0.144 ± 0.026 in the case of RCHE
powder and 0.162 ± 0.031 in the case of RCAE. These values indicate
F. Zanoni, et al.
Fig. 3. Appearance (A and B), optical microscopy images (C and D) and scanning electron microscopy images (E and F) of RCHE powder (left side) and RCAE
powders (right side).
that the obtained powders should be microbiologically stable.
The SP was 6.1 ± 0.5% in the case of RCHE and 5.7 ± 0.7% in the
case of RCAE. This information is of great importance because the su-
perficial fraction of polyphenols, that is not embedded in the en-
capsulates, is more prone to oxidation. These values can be probably
reduced adjusting spray-drying operational parameters in order to
promote a better particles formation (e.g. inlet temperature and feed
rate). Also, the increase of solids content (i.e. starch), and the inclusion
of further polymers, like maltodextrin and gum arabic, could contribute
in reducing the SP.
TP and SP allowed to calculate EE of 79.1 ± 3.0% for RCHE and
88.2 ± 1.3% for RCAE.
These data are not in accordance with the results described in Fig. 2
since, despite showing lower stability, red cabbage PP exhibited better
spray-drying retention. This higher resistance can be a consequence of
the protection exerted by OSA-starch. The capacity of PP to interact
with starch is in facts well known. In particular, it was highlighted that
the structure of PP can influence the degree and the way of interaction
with the polysaccharide (Zhu, 2015). This could be the rationale of the
different protecting behavior exerted by starch towards the two extracts
since acylated PP present in RCAE might differently interact with the
polysaccharide.
5
Food Chemistry 307 (2020) 125535
3.4. Powder stability evaluation
To understand whether the encapsulation process might have im-
proved the stability of the polyphenols, the spray-dry powders were
resuspended in water and subjected to thermal treatment at 100 °C as
described for the extracts. The results are showed in Fig. 4. The RCHE
powder showed on average 90% retainment of both PP and AOC
compared to the original extract, meaning an improved thermal stabi-
lity of 27% for PP and 20% for AOC. Similar results were obtained for
the RCAE powder, with PP and AOC% retainment of 86% and 94%
respectively. These values mean a thermal stability increment of 40%
for PP and 54% for AOC. The data clearly show that despite the original
extracts are characterized by strong differences with respect to stability
as a consequence of the different polyphenols present (anthocyanins Vs
hydroxycinnamic acids), the encapsulation process based on OSA-
starch matrix provided a protective effect leading to a substantial si-
milar stability. The results are of great importance since very frequently
the stability of the encapsulated active molecules is analyzed only in
terms of shelf life during storage at specific conditions of temperature
and humidity, leaving an important issue such as the stability of the
active during processing not addressed. This information is very im-
portant when an industrial heating process has to be deigned or tuned.
F. Zanoni, et al.
Fig. 4. Time-dependent percentage retainment of PP (A) and AOC (B) of RCHE, RCHE powder, RACE and RCAE powder upon heat treatment at 100 °C for 3 h. The
powders were resuspended in water before the treatment. The pH of the samples was 3.4–3.6.
In the present case the resuspension of the powders in water simulated a
boiling process, but it would be interesting to evaluate their perfor-
mance in relation to other kinds of processes.
3.5. Color properties of RCHE and RCAE powders
Anthocyanins-rich extracts are frequently used as natural pigments
for food and supplements applications. The encapsulation process can
influence the color properties of these pigments due to their possible
interaction with the solids added, i.e. OSA-starch, and considering the
heat treatment that the molecules undergo after atomization. Hence, it
is crucial to evaluate the color properties of encapsulated extracts. For
this reason the extracts and the resuspended powders have been buf-
fered at different pH value, from 1 to 10 in order to highlight possible
differences. In the range of pH between 1 and 3 the two extracts showed
a purple-red color peculiar of the flavylium cation structure (Fig. 5)
with a maximum of absorption at 520 nm (Fig. 6A, C). The absorbance
at this wavelength normally decreases in the pH range between 4 and 6,
due to the flavylium cation hydration that leads to the uncolored
chalcone and carbinol pseudobase forms (Francis, 1989). This was
particularly visible in the case of RCHE extract (Fig. 5), whereas for
RCAE the presence of purplish chinoidal forms was probably the reason
of the color observed in this pH range. In basic conditions (pH 7–10), a
shift in the absorbance was observed to higher wavelengths, around
580 nm for RCHE and 620 nm for RCAE (Fig. 6A and C respectively).
This bathochromic shift is a consequence of the conversion of antho-
cyanins to unstable chinoidal structures characterized by an intense
blue color in the case of RCAE (Cevallos-Casals & Cisneros-Zevallos,
2004). Differently, the exposure of RCHE anthocyanins to basic pH
values led to a simultaneous increase in the absorbance at 380–420 nm
(Fig. 6A) due to presence of chalcones and trans-chalcones
Fig. 5. Color appearance of extracts and solubilized powders at different pH
values (pH 1–10). 1: RCHE, 2: RCHE powder, 3: RCAE, 4: RCAE powder. The
powders were resuspended in water before the treatment. The pH of the sam-
ples was adjusted with HCl and NaOH.
6
Food Chemistry 307 (2020) 125535
characterized by a yellow-green color (Fig. 5, line 1). This phenomenon
is visible also for RCAE a pH 10, where the blue color observed at pH 9
shifted to yellow (Fig. 5, line 2). This was confirmed by the absorption
spectrum obtained at this pH showing a maximum at 380 nm (Fig. 6C).
In the case of RCHE powder, the absorption spectra registered at
different pH values were very similar to those of the starting extract
(Fig. 6A, B) indicating that the encapsulation process did not influence
the light-absorption properties of the anthocyanins in the visible range.
In the case of RCAE powder (Fig. 6D) a small (5 nm) hypsochromic shift
(so towards smaller wavelengths) of the maxima was observed in
comparison to the starting extract along all the pHs tested. Additionally,
the color at pH 6 appeared pinkish instead of blue (Fig. 5, line 3 vs line
4). This correlates with the spectra registered at pH 5 and 6, that in the
case of RCAE powder are perfectly overlapping (Fig. 6D) while for
RCAE they differ in the 500–700 nm range. These phenomena can be
explained considering the presence of starch that might have affected
the optical properties of the anthocyanins by interacting with their
structure, even though we cannot exclude the contribute of the high
operating temperature of spray-drying.
On the whole, the color properties of the anthocyanins were almost
unaffected by the encapsulation process. This results gives the possi-
bility to consider these powders for further applications i.e. as sensors
for food spoilage detection during storage (Pourjavaher, Almasi,
Meshkini, Pirsa, & Parandi, 2017). The release of specific molecules
during microbial growth, e.g. organic acids, ammonia, etc., could
change the color of the anthocyanins, possibly grafted to edible bio-
films, indicating the progress of the deterioration. Encapsulation might
increase the performance of these sensors since the anthocyanins are
more stable, with the possibility to work at different conditions. In
additions, it gives the possibility to confine the off-flavors/tastes de-
riving from the vegetable source, like in the case of cabbage. This would
reduce the impact on the organoleptic level increasing the acceptability
of the product by the consumer (Lavelli et al., 2017).
4. Conclusions
In the present work an extraction procedure from the edible parts of
red chicory and red cabbage was established using ethanol 70% as
solvent. The extracts were found instable if exposed to high tempera-
ture, especially in the case of red cabbage. The encapsulation process
led to an increase of the thermal stability of polyphenols and AOC of
20–30% for red chicory and of 44–55% for red cabbage. Upon micro-
encapsulation, the pH-dependent light-absorption properties of the
pigments were almost entirely preserved. These characteristics make
the microencapsulated powders here developed promising functional
ingredients and innovative pH sensors.
F. Zanoni, et al. Food Chemistry 307 (2020) 125535

Fig. 6. Visible light absorption spectra of extracts and solubilized powders at

different pH values (pH 1–10). The samples are the same presented in Fig. 5.

Declaration of Competing Interest

The authors declare that they have no known competing financial

interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.

Acknowledgments

This work was supported by the Grant of the University of Verona.

We thank Dr. Giulia Donà for her valuable technical assistance.

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