food and bioproducts processing 9 1 ( 2 0 1 3 ) 28–36

Contents lists available at SciVerse ScienceDirect

Food and Bioproducts Processing

journal homepage: www.elsevier.com/locate/fbp

Assessment of production efficiency, physicochemical

properties and storage stability of spray-dried propolis, a
natural food additive, using gum Arabic and OSA
starch-based carrier systems

Felipe C. da Silva a , Carolina Rodrigues da Fonseca a , Severino Matias de Alencar b ,

Marcelo Thomazini a , Julio C. de Carvalho Balieiro a , Paola Pittia c ,
Carmen Sílvia Favaro-Trindade a,∗
a Universidade de São Paulo – Faculdade de Zootecnia e Engenharia de Alimentos, Av. Duque de Caxias Norte, 225, CP 23, CEP: 13535
900, Pirassununga, SP, Brazil
b Universidade de São Paulo – Escola Superior de Agricultura “Luiz de Queiroz”, Av. Pádua Dias, 11, CEP: 13418900, Piracicaba, SP, Brazil
c Dipartimento di Scienze degli Alimenti, Università degli Studi di Teramo, Via C.R. Lerici 1, 64023 Mosciano S. Angelo, TE, Italy

a b s t r a c t

The aim of this work was to obtain propolis in a powder, alcohol-free, water-dispersed and shelf-stable form. Propolis
extract was spray-dried using gum Arabic and octenyl succinic anhydride (OSA) starch as carriers in two different
weight ratios (1:4 and 1:6). Spray-dried propolis samples were evaluated for morphology, moisture, water activity,
water dispersibility, hygroscopicity, particle size, particle distribution, entrapping efficiency, stability, isotherms and
antioxidant properties. The spray-drying process produced round particles with sizes ranging from 15 to 24 ␮m. This
process preserved the antioxidant activity of propolis and also allowed propolis to be obtained in a powder form,
which was stable during storage at room temperature, had low hygroscopicity and was highly dispersible in cold
water. The application of this technology could increase the use of propolis in various industrial applications, such
as an antimicrobial and as an antioxidant in food.
© 2012 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.

Keywords: Bee glue; Encapsulation; Phenolic compounds; Isotherms; Hygroscopicity; Spray-drying

1. Introduction biological and therapeutic actions are attributed to the phe-

Propolis or bee glue is based on resins collected from resinous
sprouts and exudates of some plants by Apis mellifera bees
(Moreira et al., 2008; Burdock, 1998). From a chemical point of
view, propolis is a complex product, and it has been used in
folk medicine since ancient times due to its properties (Kim
et al., 2008). In particular, recent studies have confirmed that
propolis exhibits the following pharmacological activities:
antiseptic, antimycotic, bacteriostatic, astringent, choleric,
spasmolytic, anti-inflammatory, anaesthetic and antioxidant
(Marcucci, 1995; Burdock, 1998; Banskota et al., 2001). These
nolic composition of propolis (Lahouel et al., 2004).
In recent times, the growing interest in some food indus-
tries to find natural additives has increased the efforts both
in obtaining bioactive compounds from natural raw materials
and in developing stable and functional derivative products.
The antimicrobial and antioxidant properties attributed to
propolis are thus valuable to the food industry because of its
potential effect in the retardation of lipid oxidation and its
positive effect on food product stability and shelf-life.
Propolis has potential as a natural food additive and as a
functional food ingredient, according to Mendiola et al. (2010).

∗
Corresponding author. Tel.: +55 19 3565 4139; fax: +55 19 3565 4284.
E-mail address: carmenft@usp.br (C.S. Favaro-Trindade).
Received 2 February 2012; Received in revised form 15 August 2012; Accepted 22 August 2012
0960-3085/$ – see front matter © 2012 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.fbp.2012.08.006
 food and bioproducts processing 9 1 ( 2 0 1 3 ) 28–36
The application of propolis in food, however, is still limited
because it is soluble in alcohol and has a strong taste and
aroma.
Spray-drying might be an alternative method that could
reduce these problems. Pharmaceutical and food industries
are using this technique to protect products from environmen-
tal conditions, extend product shelf life and mask unpleasant
flavours (Ré, 1998, 2006; Favaro-Trindade et al., 2008, 2010).
In a preliminary study of our research group (Silva et al.,
2011), propolis was spray-dried without a carrier agent, but
unsatisfactory results were obtained because of the low yield
of the spray-drying process, the physical instability of the pow-
der and the reduced dispersibility of the powder in water.
Bruschi et al. (2003) produced propolis microparticles
through spray-drying using gelatine as a carrier agent, and
they achieved products with entrapment efficiency of 41%. The
spray-drying technique maintained the antimicrobial activ-
ity of propolis, thereby suggesting that it can be a promising
process for developing an intermediary or eventual propolis
dosage form without ethanol or a strong, unpleasant taste.
However, the use of gelatine originating from bovine has been
reduced in recent years in some countries due the risk of
bovine spongiform encephalopathy. Thus, alternative carriers
are needed.
Gum Arabic and OSA starch could act as alternative carrier
agents to obtain spray-dried propolis products because they
exert emulsifying properties that could improve the dispersi-
bility in water.
Gum Arabic is widely used in the food industry because it
is nontoxic, odourless and tasteless. Gum Arabic is the most
widely used encapsulating material in microencapsulation by
spray-drying due to its good emulsifying and film-forming
capacities (Gharsallaoui et al., 2007) as well as its low viscosity
in aqueous solution.
Octenyl succinic anhydride (OSA) starch is a modified
starch obtained by the addition of alkenylated dicarboxylic
acid anhydride groups. As compared to native starch, OSA
starch contains both hydrophilic and hydrophobic groups in a
stable ratio of 1:1. The special structure of OSA starch is advan-
tageous in its use in foods as a surfactant. Additionally, OSA
starch has a low viscosity in aqueous solution, which also aids
the spray-drying process.
Thus, the aims of the present study were to produce propo-
lis in a powder, alcohol-free and water-dispersible form by
spray-drying using gum Arabic and OSA starch as carriers and
to study its physical and functional properties.
2. Materials and methods
2.1. Materials
Type-12 propolis, a green type according to Park’s classifica-
tion (Park et al., 2000), was used as the bioactive compound.
The propolis samples were collected in Mogi Guaçu-SP, Brazil.
The propolis ethanol extract was prepared as described by
Alencar et al. (2007) with some modifications. Approximately
30 g of propolis crushed in a blender was mixed with 100 mL
of ethanol (80◦ GL) and then placed in a water bath (Marconi,
model MA 085, Brazil) at 50 ◦ C under mechanical stirring for
30 min. The propolis ethanolic extract was used as the active
compound. Gum Arabic (GA) (CNI, Brazil) and OSA starch (Corn
Products, Brazil) were used as carrier agents.
29
2.2. Preparation of propolis carrier formulations
Individual solutions of the carriers (GA and OSA starch) were
prepared at the concentration of 30 g/100 mL. Four different
formulations (dispersions) were prepared using the different
carrier solutions (GA or OSA starch) and two propolis extract-
to-carrier solution ratios equal to 1:6 and 1:4 (w/w).
The samples made with propolis extract:GA in 1:6 and 1:4
ratios are identified in the text as T1 and T2, respectively. The
samples made with propolis extract:OSA starch in 1:6 and 1:4
ratios are identified in the text as T3 and T4, respectively.
2.3. Spray-drying conditions
The carrier agent solutions and the propolis extract were dis-
persed with a homogeniser (Ultra-Turrax T25; IKA, Germany)
for 2 min at 15,000 rpm. The formulations were then atom-
ised with a spray dryer (model MSD 1.0, Labmaq, Brazil). The
operational conditions of the spray dryer were as follows: inlet
temperature of 120 ◦ C; outlet temperature of 91 ◦ C; air flow of
0.60 m3 /min; feed flow of 0.60 m3 /min; and nozzle diameter
of 1.3 mm. These conditions were chosen based on prelimi-
nary trials and from results of previous studies of our research
group.
At the end of each drying session, the powders were gath-
ered, placed in closed vials covered with aluminium foil and
kept at room temperature in a dry and dark place until analy-
sis.
The encapsulation process and all analyses were con-
ducted in triplicate.
2.4. Samples characterisation
2.4.1. Morphology
The morphological characteristics of the microparticles were
observed by scanning electron microscopy (JEOL JSM-T300,
Tokyo, Japan) according to the method described by Oliveira
et al. (2007). The methodology described by Santos et al. (2005)
was used to break the particles.
2.4.2. Moisture and water activity (aw )
The moisture content of the samples was determined using a
moisture analyser (Ohaus, MB35, USA). The aw was measured
using a dew point hygrometer (Aqualab, Decagon Devices,
USA).
2.4.3. Water dispersibility
The method of Singh and Singh (2003) was adapted to
determine the cold water (20 ◦ C) dispersibility of the spray-
dried propolis. A suspension of 1% spray-dried propolis (w/w;
100 mL) was thoroughly shaken for 30 min on a rotary shaker
(TE-420 Tecnal, Brazil). The suspension was transferred to
a 250 mL centrifuge bottle and centrifuged at 1200 × g for
10 min. A 25 mL aliquot of the supernatant was placed in a
pre-weighed aluminium moisture dish and dried in an oven
(Tecnal, Brazil) at 110 ◦ C for 4 h. The cold water dispersibility
(CWD) was calculated as a percentage according to the follow-
ing equation:
weight (g) of solid in the supernatant × 4
CWD (%) =
weight (g) of sample
× 100.
30 food and bioproducts processing 9 1 ( 2 0 1 3 ) 28–36
2.4.4. Hygroscopicity
Aliquots of the powders (exactly 2 g each) were placed into
Petri dishes at 25 ◦ C in an airtight plastic container filled with
a saturated solution of Na2 SO4 (81% ERH). After one week, the
samples were weighed, and the hygroscopicity was expressed
as follows: g of water absorbed/100 g of dry solids (Cai and
Corke, 2000).
2.4.5. Particle size measurement
The particle size distribution was measured using a laser
light diffraction instrument (Mastersizer S, Malvern Instru-
ments, Malvern, UK). A small powder sample was suspended
in isopropanol under magnetic agitation, and the particle size
distribution was monitored during each measurement until
successive readings became constant. The particle size was
expressed as D[4,3], which is the mean diameter over the vol-
ume distribution, according to Comunian et al. (2011).
2.4.6. Total phenolic compounds and total phenolic loss
Total phenolic compounds were determined by the
Folin–Ciocalteu method, which has been described by
Woisky and Salatino (1998), using gallic acid as the standard.
A mixture of 0.05 g of the sample and 2.5 mL of Folin–Ciocalteu
reagent diluted in water in a 1:10 ratio was left to rest for
5 min. Two millilitres of 4 g/100 mL Na2 CO3 was then added,
and the mixture was allowed to rest again for 2 h in darkness.
The absorbance was then read using a spectrophotometer
(Biochrom Libra S22, Cambridge, England) at 740 nm. For
determination of the phenolic loss, total phenolic content
was calculated as g/100 g of dry matter in the mixture entering
the spray dryer as well as in the resulting powder.
2.4.7. Evaluation of the antioxidant activity
The spray-dried propolis microparticles were broken accord-
ing to the methodology described by Bruschi et al. (2003). A
sample of spray-dried propolis (15 mg) was added to 25.0 mL
of acetonitrile. After sonication for 5 min, the dispersion was
filtered through a membrane filter (pore size of 0.20 ␮m; GTTP,
Millipore).
The measurement of the scavenging ability of DPPH radi-
cals was performed according to the methodology described
by Chen et al. (2003). The reaction mixture was made by adding
1.0 mL of the 0.5 mM DPPH radical solution and 4.0 mL of the
solution that resulted from the broken microparticles. After
20 min at room temperature, the absorbance was measured at
517 nm. The antiradical activity was determined as an inhibi-
tion percentage (IP), which was calculated as the rate of decline
in absorbance of the DPPH extract solution after 20 min of
reaction (steady state) in comparison to the reference solution
(DPPH ethanol).
2.5. Stability of spray-dried propolis under storage
and critical conditions
2.5.1. Stability of the phenolic fraction
The stability test was performed by the evaluation of the total
phenolic compounds in the spray-dried samples that were
stored in glass vials, vacuum-sealed and stored at tempera-
tures of 10 and 25 ◦ C. The samples were evaluated every 30
days for 6 months.
2.5.2. Stability with regard to moisture uptake and
propolis release
The stability of the powders with regard to moisture uptake
was evaluated by the colour changes due to the propolis
release under standardised conditions mimicking potential
real application in food matrices. The differently made spray-
dried propolis powders were mixed with starch (spray-dried
propolis:starch weight ratio of 3:100) preliminarily equili-
brated at an equilibrium relative humidity percentage (ERH%)
of 84 and 91% in plastic boxes (with a diameter of 4 cm) that
were hermetically sealed. In total, eight sets of samples (2
series of samples at different ERH% values and 4 different car-
riers) were prepared. Each set (n = 8) consisted of 2 samples
with the same ERH% and carrier type. The samples were stored
at 20 ◦ C for 6 days.
The moisture uptake of the powders in direct contact with
starch at high ERH% is expected to determine the release of
the encapsulated propolis in the starch media. Due to the nat-
ural dark brown colour of the propolis, its release, induced
by the moisture uptake of the powder, determines a pro-
gressive colour change of the starch/spray-dried mix. The
colour change evaluation was performed using a colourimeter
(Minolta CR-300, Tokyo, Japan). Measurements were taken on
the surface of the powder mixture at different storage times
every 24 h. The L* parameter was chosen to describe the colour
change induced by moisture uptake by the propolis powders
and was used as an index of the stability of the spray-dried
propolis with regard to moisture uptake in the presence of
different carrier types and quantities.
2.6. Moisture sorption isotherm
The sorption isotherm was determined by the gravimetric
method. The powders were preliminarily dehydrated by expo-
sure to P2 O5 . Aliquots (1 g) were weighed in aluminium caps
and equilibrated over saturated salt solutions, providing rela-
tive humidity values in the range of 20–91.0 RH% in enclosed,
still, moist air at 20 ± 1 ◦ C. The water content was determined
from weight gain, and the sorption isotherm was constructed
at 20 ◦ C using the steady state water contents and aw val-
ues determined by an Aqualab hygrometer (Decagon Devices,
Pullman, WA).
The adsorption isotherms were described by the GAB equa-
tion as follows:
Xm · Cg · K · aw
W= (1)
(1 − K · aw )(1 − K · aw + Cg · K · aw )
where W is the water content on a dry basis; Xm is the water
content of the monolayer; aw is the water activity; and Cg and
K are constants that are related to the energies of interac-
tion between the first and further adsorbed molecules at the
individual sorption sites.
2.7. Statistical analysis
2.7.1. Characterisation of samples
To evaluate the variables, including moisture content, water
activity, dispersibility, hygroscopicity, particle size, phenolic
loss and trapping efficiency, a completely randomised design
(CRD) was adopted considering all the treatments according
to the following model: Yij =  + Ti + eij; where Yijklm is the
observed value for a physical variable (dispersibility, hygro-
scopicity, water activity, moisture content, phenolic loss and
 food and bioproducts processing 9 1 ( 2 0 1 3 ) 28–36
Fig. 1 – Micrographs of microparticles produced with: (A) GA (T1), (B) OSA-starch (T4), and (C) fragmented microparticles
produced with GA (T2) (magnification 5000×).
particle size) in repetition j of treatment I;  is the constant
inherent to all observations (average); Ti is the effect of the
ith treatment; i is equal to 1 (1 propolis:6 GA), 2 (1 propolis:4
GA), 3 (1 propolis:6 OSA starch) or 4 (1 propolis:4 OSA starch);
and eij is the experimental error associated with j repetitions
of treatment i, assuming NID (0,  and 2). Due to the sig-
nificant effects of the treatments, Student’s t-test was used
for multiple comparisons in conjunction with the previously
mentioned program.
2.8. Stability
To evaluate the variable content of phenolics during the
stability test according to the treatments used, storage tem-
peratures (10 and 25 ◦ C) and days of storage, a completely
randomised design (CRD) in a 4 × 2 × 6 factorial was adopted
according to the following model:
Yijkl =  + Ti + Rj + Dk + TRij + TDik + RDjk + TRDijk + eijkl
where Yijkl is the value observed for variable phenolics in rep-
etition l at day k of storage, refrigeration temperature j and
treatment i;  is the constant inherent in all observations
(average); Ti is the effect of the ith treatment with i = 1 (1 propo-
lis:6 GA), 2 (1 propolis:4 GA), 3 (1 propolis:6 OSA starch) or 4 (1
propolis:4 OSA starch); Rj is the effect of the jth temperature
with j = 1 (10 ◦ C) or 2 (25 ◦ C); Dk is the effect of the kth day of
storage with k = 1 (30 days), 2 (60 days) or 6 (180 days); TRije is
the effect of the double interaction of treatment i with temper-
ature j; TDik is the effect of the double interaction of treatment
i with k days of storage; RDjk is the effect of the double inter-
action of day j of storage with day k of storage; TRDijk is the
effect of the triple interaction of treatment I with temperature
j and day k of storage; and eijk is the experimental error asso-
ciated with the variable total phenolic contents in repetition l
at day k of storage, refrigeration temperature j and treatment
I, assuming NID (0,  and 2).
For both statistical models mentioned above, we proceeded
with the developments by using regression analysis within
each treatment/temperature combination because significant
effects for the triple interaction were observed. All tests were
performed with the aid of the Statistical Analysis System©
31
program version 9.1.3 (SAS, 1995) using the PROC GLM pro-
cedure.
3. Results and discussion
3.1. Characterisation of samples
3.1.1. Morphology
Spray-drying of the propolis extract with both carriers resulted
in a fine, pale yellow-coloured powder.
Fig. 1 shows the SEM microphotographs of the produced
powders. For both carriers (GA and OSA starch), the micro-
particles had a round shape, which facilitates the flow
of powder. Some microparticles showed small concavities
(dents) with an appearance that could be attributed to the
rapid evaporation of liquid droplets during the spray-drying
process (Rosenberg et al., 1985).
Independent from the type and ratio of the carrier, the
microparticles had similar appearances. The external surfaces
showed continuous walls with no fissures, cracks or interrup-
tions, which is an attribute that is essential to ensure lower
gas permeability, better protection and propolis retention.
Fig. 1C shows a fragmented particle that is hollow, indicat-
ing that the microparticles or microspheres were of a matrix
type in which the core material was distributed throughout
the carrier. This formation of microspheres was also observed
in other studies as follows: when lycopene was spray-dried
using starch (Capsul® ) (Rocha et al., 2012); when hydrolysed
casein was atomised in the presence of maltodextrin DE 10
and 20 as carriers (Rocha et al., 2009); and when hydrolysed
casein was atomised using soy protein isolate was used as
carrier (Ortiz et al., 2009).
The results for moisture content, water activity, dispersi-
bility, hygroscopicity, cold water dispersibility and particle size
are shown in Table 1.
Low moisture values are necessary to ensure the stability of
atomised powders because they prevent caking, which begins
with the agglomeration of wet particles and reduces the reten-
tion of the active principle, thereby hindering the powder flow
and dispersion.
32 food and bioproducts processing 9 1 ( 2 0 1 3 ) 28–36

Table 1 – Chemical, physico-chemical, physical and technological properties of spray-dried propolis.

Parameters Formulations

T1 T2 T3 T4

Moisture (%) 9.3 ± b

0.4 12.6 ± a
0.4 4.9 ± d
0.3 7.2 ± 0.6c
aw 0.33 ± 0.01b 0.39 ± 0.02a 0.25 ± 0.02d 0.29 ± 0.01c
Hygroscopicity (gH2 O/100 g of powder) 29.3 ± 0.6a 27.4 ± 0.9a 15.8 ± 0.9b 13.8 ± 0.1b
Cold water dispersibility (%) 86.0 ± 3.0b 84.0 ± 0.3b 94.0 ± 2.0a 86.0 ± 2.0b
Particle size (␮m) 24.0 ± 0.6a 23.3 ± 0.4b 15.0 ± 0.3d 16.0 ± 0.0c
Colour
L* 88.09 ± 0.16b 86.31 ± 0.29a 87.16 ± 0.72a 89.14 ± 0.41c
a*/b* 0.31 ± 0.01c 0.22 ± 0.01a 0.25 ± 0.03a 0.28 ± 0.03b
Phenolics loss (%) 3.4 ± 0.3a 3.0 ± 2a 9.0 ± 2b 10.5 ± 1b
Trapping efficiency (%) 85.1 ± 0.9a 76 ± 1b 86 ± 2a 78.7 ± 0.7b

Mean ± standard deviation values in the same line followed by the same superscripts (a–c) are not significantly different (P > 0.05).
T1 and T2: propolis extract:GA ratio of 1:6 and 1:4, respectively; T3 and T4: propolis extract:OSA-starch ratio of 1:6 and 1:4, respectively.

The moisture and aw values were within the expected range
for atomised products and also within the recommended val-
ues to ensure microbiological stability (<0.6). The moisture
values were similar to those obtained by Bruschi et al. (2003),
who reported values ranging from 4.12 to 9.4 g/100 g when
propolis was spray-dried with gelatine as a carrier.
The aw and moisture values of the powders had significant
differences (P < 0.05). The carrier agent type affected the mois-
ture and aw . The GA powders had significantly higher moisture
content and aw than those obtained with OSA starch. Samples
prepared with GA retained more water in the atomisation pro-
cess. This result can be attributed to the chemical structure of
GA, which has a high number of ramifications with hydrophilic
groups, resulting in a higher water binding capacity. Similar
results were also observed by Tonon et al. (2009) when they
spray-dried açai pulp using GA and maltodextrin as carriers.
With respect to water dispersibility, the microparticles
obtained with the four formulations provided high values
ranging from 84 to 94%. Fig. 2A shows that the pure or free
propolis was poorly dispersible in aqueous media and that
macroscopic flakes were visible in the bottle. In contrast,
the equivalent quantity of spray-dried propolis was fully dis-
persible in aqueous media (Fig. 2B), which was an important
result because dispersibility is one of the main constraints
Fig. 2 – Dispersibility in aqueous media of free propolis (A)
and spray-dried with OSA-starch as carrier (B).
when an additive or functional ingredient has to be used in
foods. In the case of propolis, the dispersibility increases the
potential applications in which propolis can be used as a nat-
ural additive, including applications in which the presence of
alcohol is undesirable.
Among the samples under study, the powders obtained
with OSA starch in the ratio of 1:4 (T3) showed the highest and
most significant dispersibility value (94%), and no significant
differences were observed among the other samples. These
results can be attributed to the emulsifying property of the
carriers related to their amphiphilic molecular structures with
both non-polar and polar groups. This type of molecular struc-
ture allowed them to interact simultaneously with non-polar
molecules of propolis and water, favouring the dispersion of
the propolis molecules in water. According to Gharsallaoui
et al. (2007), the technological properties of spray-dried micro-
particles depend mainly on particle/particle interactions for
flowability and particle/liquid interactions for wettability and
re-dispersibility.
Highly water-dispersible propolis particles have also been
obtained by Kim et al. (2008) using encapsulating agents
consisting of copolymers, including N-isopropylacrylamide
(PNIPAAm) with N-vinyl-2-pyrrolidone (VP) and polyethylene
glycol monoacrylate (PEG-A), obtained by a polymerisation
technique. Nevertheless, in respect to the study by Kim et al.
(2008), the advantage of the present work was related to the
use of a relatively simple and low cost technique and GRAS
food biopolymers.
According to the methodology used, the samples were
slightly hygroscopic (Table 1), which indicated that the pow-
ders are easy to store and handle. The samples atomised with
GA (T1 and T2) were significantly more hygroscopic than those
atomised with OSA starch (T3 and T4). This difference can be
attributed to the different polarities of the carriers and to the
highly branched structure of GA, making the polar interactions
with water easy by hydrogen bonds.
The spray-dried propolis microparticles had a particle size
distribution with clear bimodal behaviour, i.e., with two dis-
tinct peaks and with each one featuring a predominant size
(Fig. 3). This characteristic is beneficial in regard to powder
because small particles can penetrate in the interstices of
larger ones, making the material take up less space. The sta-
tistical analysis of the results showed significant differences
(P < 0.05) among the mean sizes of the powders with differ-
ent formulations. In general, the particles produced with GA
had a higher average diameter than those produced with OSA
 food and bioproducts processing 9 1 ( 2 0 1 3 ) 28–36 33

Table 2 – Regression parameters of GAB equation

applied to sorption isotherms.
Formulation Xm Cg K R

T1 0.052* 8.913* 0.96** 0.989

T2 0.049** 10.321* 0.986* 0.991
T3 0.047* 7.328* 0.866* 0.992
T4 0.045* 8.143* 0.907* 0.993

T1 and T2: propolis extract:GA ratio of 1:6 and 1:4, respectively; T3

and T4: propolis extract:OSA-starch ratio of 1:6 and 1:4, respectively.
∗
Significant at P < 0.05.
∗∗
Significant at P < 0.01.

related to the specific presence of hydrophilic groups in the

Fig. 3 – Particle size distribution of T1 sample (propolis two molecules, with a higher number of ramifications with
extract:AG ratio of 1:6). hydrophilic groups in the GA than in the OSA-starch, as pre-
viously discussed.
The results of the nonlinear regression analysis for fitting
starch, and this result might be attributed to the higher water the GAB model to experimental data are shown in Table 2.
retention of GA during spray drying (Table 1), which helps to The fitting of the isotherms with this model was good. In all
reduce particle shrinkage. However, the difference of approx- cases, the results obtained in the present work for the adjust-
imately 5 ␮m is not technologically significant. ment of GAB model to the different powder samples were in
The average size of particles produced was within the size accordance with the limit values for the C and K constants
range of particles produced by atomisation, which is a range suggested by Lewicki (1997) based on a mathematical analysis
from 5 to 150 ␮m, according to Thies (1995). The particles pro- of the model (range 0.24 < K < 1 and 5.6 < Cg < ∞).
duced in this study, however, were larger than those produced The Xm parameter (monolayer moisture content) corre-
by Bruschi et al. (2003), who reported a mean size ranging from sponds to the amount of water strongly adsorbed to specific
2.5 to 2.7 ␮m when gelatine was used as a carrier. This differ- sites at the food surface and is considered a critical value above
ence can be attributed to the shrinkage rate, which is highly which both the rate of some degradation reactions increases
dependent on the operating conditions of the spray dryer and and the food matrix stability decreases.
on the size of the molecule used as a carrier because smaller The Xm was higher for the GA samples than for the OSA
molecular sizes generally result in smaller particle sizes pro- starch samples. In the former powders, the data (4.8–5.2%)
duced. The particles produced in the present experiment were were close to the values reported by Tonon et al. (2009) (5.3%)
also larger than both those obtained by Durán et al. (2007), who but lower than those determined by Gabas et al. (2007) (8.1%)
reported a range from 5 to 10 ␮m when propolis was encap- and Perez-Alonso et al. (2006) (7.2%) for differently spray-dried
sulated through emulsification-solvent evaporation with a powders. With regard to the OSA starch, the Xm values of
poly(e-caprolactone) technique, and the propolis nanocap- the propolis powders were lower than those reported by Cova
sules obtained by Kim et al. (2008) (average size or 50 nm), et al. (2010) for OSA cassava starch (5.12–6.37%) when just the
who used micellar aggregates of copolymer mixtures of iso- modified native starch was considered. The degree of starch
propylacrylamide (PNIPAAm) with N-vinyl-2-pyrrolidone (VP) modification, the spray-drying conditions and the formula-
and polyethylene glycol monoacrylate (PEG-A). However, the tion could affect the amount of water strongly bound to the
microcapsules produced by these other authors most likely matrix. In both the GA and OSA starch powders, the increase
could not be used in food because synthetic polymers were of the carrier in the initial formulation led to a decrease of the
used in their preparation.
The formulation and the spray-drying operation parame-
ters used determined the achievement of pale yellow powders Table 3 – Evaluation of the antioxidant property of
spray-dried propolis.
with limited colour differences (Table 1). The powders pro-
duced with OSA starch had higher L* values than the GA Formulation Concentration Antioxidant
powders. This result could be attributed to the effect of the (ppm) activity (%)
carrier colours. 500 24 ± 1
The spray-drying conditions used in this study resulted in T1 2000 72 ± 5
a limited loss of phenolic compounds (Table 1). Among the 5000 89 ± 3
carriers tested, GA was better able to preserve the bioactive 500 33 ± 1
compounds during atomisation than OSA starch. It is possible T2 2000 79 ± 1
that the GA, which is a charged molecule, could have inter- 5000 84 ± 3
acted with the more polar phenolic compounds by providing 600 35 ± 3
a thermoprotective effect during the exposure to high temper- T3 1500 62 ± 4
atures. However, no significant effects (P > 0.05) were observed 3000 88 ± 1
due to the use of different concentrations of the carriers. 600 52 ± 1
Sorption isotherms of the differently formulated spray- T4 1500 81 ± 2
dried propolis powders were determined, and all materials 3000 87 ± 1
showed a typical sigmoidal trend, with GA propolis powders
T1 and T2: propolis:Arabic gum ratio of 1:6 and 1:4, respectively; T3
being more hygroscopic than OSA starch powders, espe-
and T4: porpolis:OSA starch at ratio of 1:6 and 1:4, respectively.
cially at aw > 0.65 (data not shown). These results could be
34 food and bioproducts processing 9 1 ( 2 0 1 3 ) 28–36
monolayer moisture content, which could be related to a dif-
ferent interaction and structuring of the molecules at the
surface of the particles when a higher concentration of the
biopolymer was present in the dispersion during the spray-
drying process (Kim and Bhowmik, 1994; Miao and Roos, 2006).
By considering the K constant in Table 2, which takes
into account the difference between the chemical potential
of the sorbate’s pure liquid state and of the upper layers
(Timmermann et al., 2001) and is in agreement with the greater
upswing of the sorption isotherm at aw > 0.65, higher val-
ues were observed in the GA powders (especially in the T1
samples) than those determined in the OSA starch powders.
This result was in agreement with the variable presence of
hydrophilic moieties in the initial biopolymer. Moreover, the
increase of the carrier content in the formulation determined
higher K values in both the GA and OSA starch formulations.
3.1.2. Antioxidant activity evaluation
Table 3 shows the results of the antioxidant activity evalua-
tion.
In general, the powders made with a lower carrier concen-
tration (T2 and T4) showed higher antioxidant activity, except
for the highest concentrations tested (3000 and 5000 ppm
for samples prepared with OSA and GA, respectively). These
results suggested that the highest concentrations were near
the saturation concentration for this activity, i.e., it was impos-
sible to work with a higher concentration to increase this
activity.
These results were in agreement with those obtained by
Marquele et al. (2006) and Souza et al. (2007), who reported
good antioxidant activity of spray-dried propolis extract, with
concentrations ranging from 2.5 to 5 mg/mL, resulting in 50%
inhibition of lipid oxidation.
3.2. Stability during storage
To consider the usefulness of spray-dried propolis for food
applications, it is important to guarantee its stability and
functionality during storage. In this context, the change of
the concentration of the phenolics was evaluated during the
storage of samples and was used as an index of the powder
stability.
Fig. 4 shows the change of total phenolic content during
sample storage for 180 days at 10 and 25 ◦ C.
With regard to the contents of total phenolics determined
during the stability test, significant effects were observed
(P < 0.05) for the source of variation related to the formula-
tion × storage temperature × storage time interaction. In this
case, regressions within each formulation × temperature com-
bination were adjusted because the effect of storage time was
considered a quantitative factor.
The formulation × temperature combinations of T1 – 10 ◦ C,
T2 – 25 ◦ C, T3 – 10 ◦ C and T4 – 25 ◦ C behaved in a decreasing
linear trend. However, the formulation × temperature com-
binations of T2 – 10 ◦ C, T2 – 25 ◦ C and T4 – 10 ◦ C showed a
quadratic behaviour. For the T3 – 25 ◦ C combination, the phe-
nolic content remained unchanged throughout the storage
period, and no definite behaviour pattern was identified.
Fig. 4 shows that the T1 and T3 samples at both storage
temperatures provided the highest stability for the phenolic
compounds, which corroborated with the angular coefficients
of the curves obtained. Both T1 and T3 were characterised by a
higher carrier content in their formulation (1:6), which could
be related to the greater protection against degradation for
Fig. 4 – Content of total phenolic compounds in the
samples T1 () (y = 0.8951 − 0.00156x), T2 ()
(y = 1.6405 − 0.01285x + 0.000041x2 ), T3 ()
(y = 1.0773 − 0.00108x), T4 (䊉)
(y = 1.1785 + 0.00123x − 0.00002x2 ) during the storage at
10 ◦ C(±1) and in the samples T1 () (y = 0.8539 − 0.00129x),
T2 () (y = 1.5439 − 0.01062x + 0.000033x2 ), T3 (♦) (y = 0.9737),
T4 () (y = 1.3565 − 0.00264x) during the storage at
25 ◦ C(±1). T1 and T2: propolis extract:GA ratio of 1:6 and
1:4, respectively; T3 and T4: propolis extract:OSA-starch
ratio of 1:6 and 1:4, respectively.
the total phenolic compounds. This result was also supported
by the fact that these powders showed a higher entrapping
efficiency compared to the other powders (Table 1), i.e., there
was less free propolis on the surface of the particles, which is
characteristic of the greater physical protection ability of the
carrier.
The storage temperature (10 and 25 ◦ C) did not affect the
stability of phenolic compounds in propolis, thereby suggest-
ing that these materials can be stored at room temperature.
These results were in agreement with those reported by Nori
et al. (2011), who did not observe a temperature effect on the
stability of encapsulated propolis by complex coacervation
using isolated soy protein and pectin as encapsulating agents.
Fig. 5 shows that the L* colourimetric parameter of the
spray-dried propolis in the presence of different carrier types
and concentrations mixed with starch equilibrated at 84 and
91 ERH% changed as a function of time.
The different formulated spray-dried propolis samples
showed a different stability upon moisture uptake. At 84
ERH%, all samples lost their free-flowing behaviour, with the
formation of small blocks resulting from the compaction and
caking of the rehydrated dried propolis in the starch medium.
However, the samples with OSA starch did not show any
changes in the colour of the medium, and the samples made
with GA, in particular the ones mixed with the T1 samples,
showed a significant L* decrease that could be related to a
destabilising moisture uptake of the amorphous dried carrier,
causing its phase transition and eventual release of the inner
propolis.
For the samples mixed with the starch equilibrated at 91%
ERH, the propolis dried in the presence of OSA starch at the
ratio 1:6 (T3) was more prone to the destabilising effects of the
plasticisation effect of water due to its uptake from the starch
with a higher L* decrease (P < 0.05) than that observed in T1.
At day 7 (168 h), some liquid-like brown spots could also be
noticed. This effect was not observed in the case of the other
spray-dried propolis equilibrated under the same ERH even if
a decrease of the L* parameter had occurred, and the effect
 food and bioproducts processing 9 1 ( 2 0 1 3 ) 28–36 35

A Propolis atomised with GA and OSA starch can be stored

95 at room temperature because it was stable during storage,
90
especially when using a propolis:carrier ratio of 1:6.
Using a treatment containing GA at a ratio of 1:6 is the best
85
method to atomise propolis.
80
L*

75
Acknowledgements
70

65 The authors thank FAPESP for the financial support (Process

60 07/00575-2) and for scholarship conceded (Process 07/007305-
0 20 40 60 80 100 120 140 160 180 0).
time (h)

B References
95 T1
T2
90 T3
Alencar, S.M., Oldoni, T.L.C., Castro, M.L., Cabral, I.S.R.,
T4 Costa-Neto, C.M., Cury, J.A., Rosalen, P.L., Ikegaki, M., 2007.
85
Chemical composition and biological activity of a new type of
80 Brazilian propolis: red propolis. J. Ethnopharmacol. 113,
L*

75
70
65
60
0 20 40 60 80 100 120 140 160
time (h)
Fig. 5 – Change of the L* of propolis powders made with
different carriers mixed with starch equilibrated at (A) 84
and (B) 91% ERH as a function of time. T1 and T2: propolis
extract:GA ratio of 1:6 and 1:4, respectively; T3 and T4:
propolis extract:OSA-starch ratio of 1:6 and 1:4, respectively.
was of the same entity of the samples stored in the starch
equilibrated at 84% ERH.
These results suggested that the significant moisture
uptake occurring at 91% ERH could lead to a macroscopic dis-
solution of the carrier and to a related release of the propolis
only in the T3 spray-dried propolis, which agreed with its
higher hygroscopicity (Table 1), thus indicating a higher affin-
ity for water and related effects on the sample stability.
In the other dried propolis samples, the plasticisation effect
of water related to 91% ERH could only induce swelling and
favour a partial release of the encapsulated propolis, which
was indicated by a reduced L* decrease in the starch system.
The physical changes observed in these samples stored
at high ERH% values were in agreement with those reported
by Tonon et al. (2009) in spray-dried acaí juice in the pres-
ence of tapioca starch, maltodextrins and GA. However, these
researchers did not observe liquefaction in the starch-made
powders, but they observed liquefaction in the powders con-
taining maltodextrin and GA. The type of starch and the
spray-drying conditions could be related to these differences
in the results of the stability of the powders.
4. Conclusions
The atomisation process made it possible to obtain propo-
lis in a powder form that was dispersible in cold water, thus
increasing the possibility of using it as a functional ingredient
or additive in various products, including food.
The samples had particle sizes, water activity and mois-
ture content suitable for atomised materials while still being
slightly hygroscopic and highly dispersible in cold water.
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