Food Chemistry 197 (2016) 1280–1285

Contents lists available at ScienceDirect

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

Review

Stabilization of betalains: A review

Mohammad Imtiyaj Khan
Department of Biotechnology, Gauhati University, Gopinath Bordoloi Nagar, Guwahati 781014, India

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

Article history: Betalains are vacuolar pigments composed of a nitrogenous core structure, betalamic acid [4-(2-oxoethy
Received 13 June 2015 lidene)-1,2,3,4-tetrahydropyridine-2,6-dicarboxylic acid]. This compound consists of a chromophore
Received in revised form 18 October 2015 substructure 1,7-diazaheptamethin. Betalamic acid condenses with imino compounds (cyclo-DOPA or
Accepted 10 November 2015
its glucosyl derivatives), or amines and/or their derivatives to form a variety of betacyanins (violet)
Available online 11 November 2015
and betaxanthins (yellow), respectively. These pigments exhibit absorption maxima in between
532–550 nm and 457–485 nm, respectively. These colors are currently in use as food additives as they
Keywords:
are bioactive and completely safe to consume. However, owing to poor stability, their potential
Betacyanins
Betaxanthins
application in pharmaceuticals and cosmetics is severely compromised. There is a lack of scientific
Complex formation reports highlighting their superior tinctorial strength including fluorescence, and their excellent water
Encapsulation solubility. For widening the color spectrum and their usage, various betalain-rich extracts have been
Copigmentation reviewed here, focusing on their matrix effects on stability vis-a-vis purified pigments of different struc-
tural make-up. This review also encompasses work carried out in the past related to stability/stabilization
of betalains and future research direction for the same.
Ó 2015 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1280
2. Stabilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1281
2.1. Complex formation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1282
2.2. Copigmentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1283
2.3. Encapsulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1283
3. Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1284
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1284
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1284

1. Introduction (protonated 1,2,4,7,7-pentasubstitued 1,7-diazaheptamethin sys-

There are about 75 betalains structurally and unambiguously
identified from plants belonging to more than 15 families classed
under Caryophyllales, wherein they serve as chemosystematic
markers (Khan & Giridhar, 2015). Betalains are made up of beta-
lamic acid [4-(2-oxoethylidene)-1,2,3,4-tetrahydropyridine-2,6-di
carboxylic acid], a nitrogen containing pigment core structure
E-mail addresses: imtiyaj@hotmail.com, imtiyaj@gauhati.ac.in
tem) (Fig. 1a). Spontaneous condensation of betalamic acid with
cyclo-DOPA or its glucosyl derivatives, and amines and/or their
derivatives gives rise to violet betacyanins and yellow betaxan-
thins, respectively (Fig. 1b and c). The absorption (kmax) in water
of betalamic acid, betaxanthins, and betacyanins is 424 nm,
471.5 ± 13.5 nm, and 541 ± 9 nm, respectively (Khan & Giridhar,
2015). The presence of ‘1,7-diaza’ substructure prompted some
researchers to claim betalains as chromoalkaloids (Reznik, 1981),
but their stability in slightly acidic pH disproved the claim

http://dx.doi.org/10.1016/j.foodchem.2015.11.043
0308-8146/Ó 2015 Elsevier Ltd. All rights reserved.
 M.I. Khan / Food Chemistry 197 (2016) 1280–1285
a b
O R2O
H H
HOOC N COOH
H 19
Betalamic acid
R1= H, R2= H;
c HO
H R
+
N
HO
R1= HO
2
3 OH
HO
4 HO
H R1= HO
HOOC
HOOC 5 N COOH HO
H HO
R= amino acid, amine or
derivatives: Betaxanthins
(32 structures) R1= H, R2= Glucosyl or derivatives; Gomphrenin group (4 structures)
Fig. 1. Representative structures of betalains.
(Delgado-Vargas, Jiménez, & Paredes-López, 2000). Betalains show
greatest stability in the pH range 4–6 and if stored at 4 °C. Hence,
these pigments could be used as colorants in frozen foods, low
temperature dairy products and short-shelf life foods. However,
these pigments are prone to various physicochemical factors, such
as light, heat, high pH (>6), air and metals, such as Fe3+, Fe2+, Cu2+,
Cu+, Sn2+, Al3+, Hg2+, Cr3+ (reviewed by Herbach, Stintzing, & Carle,
2006). Due to this, betalains have limited applications in foods. On
the other hand, betalains could be stabilized to a certain extent by
antioxidants, such as ascorbic acid (AA), isoascorbic acid, citric
acid, and other preservatives, whereas antioxidant phenolic com-
pounds have no stabilization effect on the pigments because degra-
dation of betalains does not involve free radical formation (Attoe &
von Elbe, 1985; Pasch & von Elbe, 1979). Organic acids, such as lac-
tic acid, acetic acid and even betalamic acid (the chromophore of
betalains), tend to dissociate completely at pH P 5 and act as
nucleophiles (Tutone, Lauria, & Almerico, 2015). As a result of this,
the presence of organic acid may either enhance the first order
degradation process of betalains by carrying out a nucleophilic
attack on N 1 (Kearsley & Katsaboxakis, 1980), or produce no sta-
bilizing effect. However, it appears that glucose oxidase can reduce
pigment degradation, particularly through oxidation, by removing
dissolved oxygen from the medium (Mikova & Kyzlink, 1985). In
addition, oxalic acid may protect betalains from digestion in the
GI tract (Eastwood & Nyhlin, 1995). Betalain stability was
improved when mixed with b-cyclodextrin (Drunkler, Fett, &
Bordignon Luiz, 2006), probably due to the formation of a 1:1
inclusion complex (Norasiha, Mimi Sakinah, & Rohaida, 2009).
Similarly, maltodextrin also could stabilize betacyanins, and indi-
caxanthin through encapsulation carried out by spray-drying (Cai
& Corke, 2000; Gandía-Herrero, Jiménez-Atiénzar, Cabanes,
García-Carmona, & Escribano, 2010). From the information
provided above, it is clear that besides antioxidants, inclusion com-
plexes and microencapsulation, metal chelating agent EDTA also
stabilizes betalains by preventing metal-induced bleaching
through the formation of an EDTA–metal complex (Attoe & von
Elbe, 1985). Literature pertaining to the chromatic stability of
1281
4
R1O 5 9 3
H
6 8 N+ 2 COOH
7 10
11
12
13
14 18
16
HOOC 15 N 17 COOH
H 20
Betacyanins
Betanidin
O
(glucosyl) or derivatives, R2= H;
Betanin group (17 structures)
Bougainvillein-r group (4 structures)
O
(2-glucuronyl glucosyl) or derivatives, R2= H;
Amaranthin group (5 structures)
O O
OH
Bougainvillein-v group (11 structures)
betalains has been reviewed (Azeredo, 2009; Delgado-Vargas
et al., 2000; Herbach, Stintzing, et al., 2006; Stintzing & Carle,
2007, 2008). The focus of future research should be on the stabi-
lization of betalains as these pigments remain under-utilized in
many industries, such as pharmaceuticals and cosmetics, owing
mainly to poor stability. So far, there is no review of the reports
on stabilization techniques employed for betalains. This review
encompasses the literature on stabilization of betalains in order
to highlight the positive aspects of betalain stability.
2. Stabilization
Betanin (EEC No. E162), approved as a red food colorant by the
European Union (Downham & Collins, 2000) and under Sec-
tion 73.40 in chapter 21 of the Code of Federal Regulations (CFR),
part of the Food and Drug Administration (FDA) in the USA
(Griffiths, 2005), has been in use as a colorant for food products,
such as hard candies, yogurts, ice creams, salad dressings, ready-
made frostings, cake mixes, meat substitutes, powdered drink
mixes, gravy mixes, marshmallow candies, soft drinks and gelatin
desserts (Delgado-Vargas et al., 2000). In most food colorant appli-
cations, an estimated quantity of less than 50 mg betanin/kg could
produce the desired color (Delgado-Vargas et al., 2000). The usages
of betalains have been greatly reduced owing to its poor stability,
which poses a significant problem when competing with synthetic
dyes (Ozela, 2004). A list of factors that reduce/enhance betalain
stability is presented in Fig. 2a. Manchali, Murthy, Nagaraju, and
Neelwarne (2013) listed the possible changes that betalains may
undergo during degradation, such as breakdown of the aldimine
bond, dehydrogenation, deglycosylation, decarboxylation and
isomerisation (Fig. 2b). In order to increase its commercial applica-
tions, betalain pigments need to resist or slow down these changes
through stabilization techniques. Among the various ways of
stabilizing betalains, complex formation, copigmentation, and
encapsulation are presented as promising techniques.
1282 M.I. Khan / Food Chemistry 197 (2016) 1280–1285
a shown that b-CD forms a 1:1 inclusion complex with betanin
b Sevilla, López-Nicolás, Carbonell-Barrachina, & García-Carmona,
R O
HO O O
HO
H AlO, and zeolite for betalain adsorption and sequestration. It was
OH
+ observed, through sophisticated analytical tools, such as NMR
N
HO COOH
H
HOOC N COOH
H under acidic conditions with the carbonyl groups of the betalain
Fig. 2. Factors affecting betalain stability (A) and sites prone to deglucosylation
(blue bracket), decarboxylation (green), dehydrogenation (red) and nucleophilic
attack (yellow bracket) in betacyanins (B). Updated from Herbach, Stintzing, et al.
(2006) and Manchali et al. (2013). PPO – polyphenol oxidase, POD – peroxidase. (For
interpretation of the references to color in this figure legend, the reader is referred
to the web version of this article.)
Fig. 3. Drug–cyclodextrin binding in equilibrium to form a 1:1 inclusion complex.
Adapted with modification from Stella and He (2008).
2.1. Complex formation
A bathochromic shift in color absorption has been observed,
indicating complex formation of betalains (especially betanin)
with metal ions, such as Cu2+ (Butera et al., 2002; Kuusi, Pyysalo,
& Pippuri, 1977), Cu+, Hg2+ (Kuusi et al., 1977), Fe2+, Ni2+, Co2+
(Spórna-Kucab, Wybraniec, Mitka, Kowalski, & Michalowski,
2011), and has been confirmed by the addition of metal-chelator
EDTA, which could reverse the spectral changes. It appears that
such complexes reduce the color stability of betalains as in the case
of Fe2+, Cu2+ (Khan & Giridhar, 2014; Pasch & von Elbe, 1979;
Spórna-Kucab et al., 2011), and Ni2+, Co2+ (Spórna-Kucab et al.,
2011). On the other hand, the b-cyclodextrin (b-CD)-betanin com-
plex has been reported to stabilize color (Hamburg & Hamburg,
1991). This was attributed to the adsorption of free water by
b-CD. In the absence of light and O2, CDs have been reported to
improve betalain stability (Drunkler et al., 2006). Recently, it was
(Norasiha et al., 2009), which was confirmed through IR spec-
troscopy and X-ray diffraction studies. Notably, the inclusion com-
plex was clearly different from the 1:1 physical mixture. a, b, c-CD
are oligosaccharides containing six to eight D-glucopyranose resi-
dues connected by a-1,4-glycosidic bonds. These macromolecules
are arranged in a toroidal shape wherein the central cavity is
hydrophobic, and the outer surface is hydrophilic with exposed
hydroxyl groups. It has been proposed that a drug/biomolecule
can sit in the cavity forming an inclusion complex with CD
(reviewed by Stella & He, 2008) (Fig. 3), both in the solid and solu-
tion phase. The review also mentioned that among the CDs,
hydroxypropyl-b-CD (HP-b-CD) and sulfobutylether-b-CD (SBE-b-
CD) are not toxic in vivo. Therefore, HP-b-CD and SBE-b-CD appear
to be promising betalain stabilizing agents for food and other
applications. Additionally, CDs improve global sensorial quality
without reducing the aroma of the sample/extract (Andreu-
2011).
Recently, a few interesting studies on inorganic matrices as sta-
bilizing agents of betalains have been published. Lima et al. (2009)
studied various matrices, including c-alumina, Mg-AlO, Mg, Zn-
spectroscopy, that adsorption of betalains in c-alumina could sta-
bilize the pigment for more than twenty months through acid–
base interaction. In another study, ceramic particles (tetraethyl
orthosilicate, TEOS) have been proposed for the stabilization of
betalains against UV-light, and temperature to increase its applica-
tion (Estévez, Méndez, Vargas, García, & Rodríguez, n.d.).
TEOS was reported to form new carbonyl-oxygen-silicon bonds
molecule (Molina, Coulson, Cortez-Valadez, García-Hernández, &
Estevez, 2014). The new complex was relatively more stable in
UV, pH and temperature, apparently by quenching UV-Vis photons,
delaying structural changes of betalains in different pH, and heat
dissipation to mask the effect of temperature on betalains. The
new complex was characterized by using FTIR spectroscopy, which
revealed the appearance of a band at 530 cm 1 possibly indicating
a C-O-Si group vibration signal in agreement with a hypothetical
structure of the new complex generated using a software program
and authenticated by simulating IR spectra. As a result of the new
structure formation, there was either shift or disappearance of
some of the signals corresponding to carboxylic acid group in the
unmodified pigment. In the case of Rivina humilis berry extract, a
possible ascorbic acid-Se-betalain complex has been reported
(see Section 2.2 for details; Khan & Giridhar, 2014).
Phenolic acids, such as gallic acid (Khan & Giridhar, 2014),
chlorogenic acid, and flavanols like catechin, quercetin, and buty-
lated hydroxyanisole have earlier been found to have either no
effect, or to reduce stability of betanin (Attoe & von Elbe, 1985).
In contrast, stabilization of betalains in the presence of flavans
has been proposed in a patent (Tsai & Hsiao, 2010). The inventors
claimed that 0.005 M catechin was sufficient to stabilize betalains
in solution, and that after heat treatment during regeneration of
the pigments a relatively stable Schiff base complex was formed
between the betalains and catechin. However, this study had some
drawbacks as the concentration of pigment was not clear, and con-
sequently the quantity of the free betalains and catechin–betalain
complex remained unclear. This information is important, particu-
larly because the relative abundance of ions in the mass spec-
troscopy analysis was very low. Also, only one concentration of
the stabilizer was used. Complex formation is poorly supported
by the analytical data. On the other hand, the observations may
be due to a copigmentation phenomenon, which does not neces-
sarily involve complex formation.
 M.I. Khan / Food Chemistry 197 (2016) 1280–1285
2.2. Copigmentation
Willstatter and Zollinger (1916) discovered the copigmentation
phenomenon in grape pigment, oenin (later named as malvidin
3-glucoside), when its hue changed to a bluer red, following the
addition of tannin or gallic acid. Later, Maud and Robert (1931)
reported the same pigment to produce diverse color hues in differ-
ent plant parts. Copigmentation involves direct (complex forma-
tion) or weak interactions between anthocyanins and other
naturally occurring compounds, causing a wide range of colors to
be expressed. Copigments could be metals, phenols, alkaloids,
amino acids, organic acids, nucleotides, polysaccharides, metals,
and anthocyanins themselves (Mazza & Brouillard, 1990). Through
copigmentation, color intensity and stability of anthocyanins are
enhanced naturally. The increase in color intensity of anthocyanins
is attributed to a bathochromic shift in its absorption spectrum and
concomitant hyperchromaticity (Mazzaracchio, Pifferi, Kindt,
Munyaneza, & Barbiroli, 2004). In the case of betalains, stabilizers,
such as ascorbic acid, isoascorbic acid, and citric acid, have been
known to remove O2 from the solution and reduce the polarity at
the N 1 position, which is susceptible to nucleophilic attack by
water (Herbach, Stintzing, et al., 2006; Kearsley & Katsaboxakis,
1980). During structure elucidation of betacyanins in Gomphrena
globosa (Heuer, Wray, Metzger, & Strack, 1992), it was observed that
acylation in gomphrenin I resulted in a bathochromic shift of
absorption (kmax) from 543 nm to 550 nm for gomphrenin II and
III. For betanin type betacyanins, esterification with aliphatic acids
stabilizes them (Herbach, Stintzing, et al., 2006), whereas if aro-
matic acids are involved in the esterification with betacyanins,
the position of esterification determines the stability as in bougain-
villeins (Heuer et al., 1994; Schliemann & Strack, 1998). Apart from
this, glucosylation at C-6 of betanidin seems to enhance the chro-
matic strength of the pigment, as in the case of gomphrenins, owing
to a more rigid conformation. However, further glucosylation may
not improve the stability (Huang and Von Elbe, 1986). Recently, it
was observed that the presence of Se and ascorbic acid enhances
stability and color intensity (hyperchromaticity) owing to a batho-
chromic shift of the betanin absorption spectrum from 535 nm to
544 nm in R. humilis fruit juice (Khan & Giridhar, 2014). It is plausi-
ble that the observed changes may be attributed to formation of an
ascorbic acid (copigment)–Se–betanin coordinate complex similar
to that of the ascorbic acid–metal–anthocyanin complex (Sarma,
Sreelakshmi, & Sharma, 1997). The natural matrix of plant extracts
contain various co-pigments that can help in stabilization and
regeneration. In the case of cactus fruit juice, autooxidation of beta-
lains occurred less in the matrix (Merin, Gagel, Popel, Bernstein, &
Rosenthal, 1987), repeated thermal treatment did not result in sig-
nificant decolorisation or non-enzymic browning (Moßhammer,
Stintzing, & Carle, 2005), and also there was some sort of synergistic
effect on the stability of betalains observed in the presence of the
juice matrix and additives compared to the matrix-free sample
(Herbach, Rohe, Stintzing, & Carle, 2006; Moßhammer, Rohe,
Stintzing, & Carle, 2007; Skopinska, Szot, & Wybraniec, 2015).
Similar results were obtained in the case of R. humilis berry juice
betacyanins and partially purified betacyanins (Khan & Giridhar,
2014). The authors further reported that in the presence of ascorbic
acid and selenium, the stability of betacyanins was much more
enhanced at 5 °C and 90 °C. The spectral profile of partially purified
betacyanins acquired before and after thermal treatment at 90 °C,
or storage at 25 °C for about 3 weeks, revealed a bathochromic shift
of 9 nm in betanin’s absorption, indicating the possibility of com-
plex formation (Khan & Giridhar, 2014). This showed that in addi-
tion to stabilization, copigments/stabilizers could bring about
chemical transformation of the pigment through simple interven-
tions, such as thermal treatment. In a complex food matrix (of fruit
spread prepared from banana) formulated using R. humilis berry
1283
juice as the natural colorant in the presence of citric acid and
sodium benzoate as preservatives, the color retention was 68%
after 6 months of storage at 5 °C (Khan et al., 2015). This was
comparable to that of betalain stability in R. humilis berry juice dur-
ing cold storage, thereby suggesting the suitability of betalains as a
colorant in a complex food matrix. Most of these studies indicated
better stability of betalains in a juice matrix compared to purified
pigments either with or without externally added stabilizers. More
studies are needed on copigments of betalains in the near future to
evaluate whether stabilization of betalains affects the bioavailabil-
ity of the pigments.
2.3. Encapsulation
Encapsulation has been studied in recent times as a method to
stabilize, improve bioavailability and ease administration of
polyphenols (Fang & Bhandari, 2010; Krishnaiah, Nithyanandam,
& Sarbatly, 2014). In the food industry, encapsulation employing
spray-drying technology is economical, flexible, could be operated
continuously, and yields good quality capsules. The process is typi-
cally used for producing encapsulated phytochemicals which are
dry, stable and used as food colorants, additives, and flavors. The
process involves mixing of the core material with a wall/coating
material. The homogenized mixture of core material and wall mate-
rial is fed into a spray dryer. Through a nozzle, atomized droplets are
sparged, the water in the droplets is evaporated by circulating hot
air, and spherical capsules of mean size range 10–100 lm are
obtained (Fang & Bhandari, 2010). With respect to encapsulation
of betalains, it was suggested that use of polysaccharides, such as
pectin, or guar gum as wall materials reduced hygroscopicity,
thereby increasing stability (Lejeune, Pouget, & Pourrat, 1983). Fur-
ther exploration in this area was done in the year 2000 wherein
Amaranthus betacyanins were encapsulated (Cai & Corke, 2000)
using maltodextrin (MDE, 10–25 dextrose equivalent), and
native/modified starch as the coating agent in a feed mixture of
20% total solids. They observed that higher inlet temperature
(>180 °C) caused increased drying loss (>4%) of pigment and
reduced storage stability. The authors assumed a superior storage
stability of encapsulated pigments with MDE of mixed dextrose
equivalent (DE) based on the observation that there was a loss of
10–16% color, which was relatively less, over a four-month storage
period. It was confirmed by encapsulation with MDE (10 DE) of
Opuntia lasiacantha Pfeiffer (red prickly pear) betanin extract, result-
ing in a loss of only 14% over a storage period of 6 months in dark at
25 °C (Díaz, Santos, Kerstupp, Villagómez, & Scheivar, 2006). In the
case of red beet betalains, encapsulation protected the pigments
independently of MDE concentration, thereby limiting the degrada-
tion to only 10% during six months of storage at 27 °C (Azeredo,
Santos, Souza, Mendes, & Andrade, 2007). These observations were
further supported by similar results obtained by different research-
ers (Obón, Castellar, Alacid, & Fernández-López, 2009; Saénz, Tapia,
Chávez, & Robert, 2009). Saenz et al. (2009) reported that indicaxan-
thin in encapsulated extract was more stable than betacyanin at
60 °C. This was corroborated by Gandía-Herrero, Jiménez-Atiénzar,
Cabanes, García-Carmona, and Escribano (2010) who encapsulated
purified indicaxanthin successfully with MDE (20%, w/v) to stabilize
the pigment without compromising the color intensity. They
observed that there was no significant loss of encapsulated pigment
during storage in the dark at 4 °C and 20 °C for more than six
months. From the above reports it appears that stability of the
encapsulated pigment depends upon the type of pigment viz. beta-
cyanins or betaxanthins. Also, these reports point to the fact that
encapsulation has great promise for the hygroscopic and poorly
stable betalains to widen its commercial applications through stabi-
lization. In this regard, Pietrzkowski and Thresher (2010) patented a
formulation of MDE (20%) and red beet dried extract with 15% (w/w)
1284 M.I. Khan / Food Chemistry 197 (2016) 1280–1285
betalains in a free-flowing product containing 5% (w/w) betalains,
which was stable and highly soluble in water. Apart from MDE,
gum Arabic has been also used as an encapsulating agent (Pitalua,
Jimenez, Vernon-Carter, & Beristain, 2010). There is a possibility of
inclusion of certain additives that can confer better functionality
to encapsulated betalain pigments as reported by Chik, Abdullah,
Abdullah, and Mustapha (2011). There are sources of betalains, such
as Basella rubra and Opuntia ficus-indica fruits, that contain natural
gummy substances, which if extracted and/or reconstituted with
pigments, may act as an intrinsic agent for stability as well as
value-addition by imparting bioactivities, such as detoxification,
digestion (USDA SR23, 2010) and improvement in hematological
parameters (Bamidele et al., 2010), which need further
investigation. The cactus cladode mucilage mixed with MDE (DE
20) could also be an encapsulating agent (Otálora, Carriazo,
Iturriaga, Nazareno, & Osorio, 2015) with functional values of the
mucilage (El-Mostafa, El Kharrassi, Andreoletti, Vamecq, & El
Kebbaj, 2014). Gelatin-MDE (Castro-Munoz, Barragan-Huerta, &
Yanez-Fernandez, 2014) and CapsulÒ (Vergara, Saavedra, Sáenz,
García, & Robert, 2014) have also been tested for improving
encapsulating efficiency, pigment retention, storage stability and
antioxidant activity of the microparticles. Whatever the strategy
used to retain maximum pigment during spray-drying, the
processed betalain powder should be acceptable to consumers.
However, a recent report points to poor consumer response, though
storage stability at temperatures up to 20 °C was impressive
(Kaimainen, Laaksonen, Järvenpää, Sandell, & Huopalahti, 2015).
Of all the drying processes, spray-drying takes the shortest dry-
ing time. The drying time of droplets from the slurry is determined
by operating parameters, such as rate of flow, pump rate, aspira-
tion rate, and inlet temperature. After reviewing the literature on
betalain encapsulation, the optimum conditions for spray-drying
of MDE (DE  20) + betalain/betalain-rich extract are: feed mixture
(20% total solids), betalains:MDE (1:3), inlet temperature,
140–150 °C; outlet temperature, 65–70 °C; feed rate, 4–5 ml/min;
air flow, 5–6 l/min; drying air flow, 40–45 m3/h or 20 psi. Through
this operation, the yield of encapsulated pigment could be any-
where between 90–98% and with a moisture content less than
5%. Moisture content or aw value is directly related to cleavage of
the aldimine bond of betalains as aw increases mobility of reactants
and oxygen solubility as well as nucleophilic attack at N 1
(Herbach, Stintzing, et al., 2006; Kearsley & Katsaboxakis, 1980).
Therefore, the aw value should be below 0.6 to improve betalain
stability (Kearsley & Katsaboxakis, 1980).
The yield and moisture content of spray-dried betalain powder
appears to decrease with increase in inlet temperature. By compro-
mising the efficiency of the drying process and stability of betalain
pigments, Gandía-Herrero, Cabanes, Escribano, García-Carmona,
and Jiménez-Atiénzar (2013) could successfully encapsulate and
co-encapsulate betanidin and miraxanthin V. For carotenoids, such
as b-carotene and bixin, 62% (Desobry, Netto, & Labuza, 1997;
Loksuwan, 2007), and 54% (Barbosa, Borsarelli, & Mercadante,
2005) yields have been reported, respectively. Anthocyanin encap-
sulation yields ranging from 80% to 97% (Ferreira, Faria, Grosso, &
Mercadante, 2009), and 65% to 93% in the presence of additives,
such as acacia gum and tricalcium phosphate, have been reported
(Nayak & Rastogi, 2010). Similarly, betalain recovery after spray-
drying could be improved by using MDE mixed with xanthan
gum as encapsulating agent (Ravichandran et al., 2014). The
authors also observed that compared to spray-drying, freeze-
drying was more effective at improving betalains recovery after
encapsulation. However, there are only a few reports pertaining
to freeze-drying of betalains. This is an important area to be
explored because freeze-drying could be the best method of
dehydration of fruit juice and foodstuffs without compromising
the color and nutritional value provided by the matrix.
3. Conclusions
In 2009, the global demand for food color was 45,000–50,000
tons (Manchali et al., 2013) and it may be assumed that about
10% of the demand was fulfilled by betalains obtained from red
beet (Hilpert & Dreiding, 2007). Recent developments in the meth-
ods for stabilization and production of free-flowing betalain pow-
der will help it to claim a bigger share in the natural color
market for applications in the food, pharmaceuticals and cosmetic
industries. However, more studies on mixed matrix based betalain
formulations and its freeze-drying as a means of drying are
required as spray-dried betalain powder elicits poor consumer
response (Kaimainen et al., 2015). From a bioavailability and meta-
bolism point of view, preservatives like oxalic acid may result in
reduced consumer acceptability owing to beeturia, a physiological
aberration in 14% of the population to process betalains (Eastwood
& Nyhlin, 1995; Mitchell, 2001). On the other hand, complexation
of betalains with metals, cyclodextrins, etc may improve stability
and, hence, their effects on betalain bioavailability need to be
investigated.
Acknowledgements
M.I. Khan is grateful to CSIR, New Delhi for awarding him a
senior research fellowship. This review was written as a part of
Mr. Khan’s doctoral thesis.
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