Journal of Microencapsulation

Micro and Nano Carriers

ISSN: 0265-2048 (Print) 1464-5246 (Online) Journal homepage: http://www.tandfonline.com/loi/imnc20

Hot-melt extrusion microencapsulation of

quercetin for taste-masking

Chia Miang Khor, Wai Kiong Ng, Parijat Kanaujia, Kok Ping Chan & Yuancai
Dong

To cite this article: Chia Miang Khor, Wai Kiong Ng, Parijat Kanaujia, Kok Ping Chan & Yuancai
Dong (2017) Hot-melt extrusion microencapsulation of quercetin for taste-masking, Journal of
Microencapsulation, 34:1, 29-37, DOI: 10.1080/02652048.2017.1280095

To link to this article: https://doi.org/10.1080/02652048.2017.1280095

Accepted author version posted online: 09

Jan 2017.
Published online: 23 Jan 2017.

Submit your article to this journal

Article views: 137

View Crossmark data

Citing articles: 2 View citing articles

Full Terms & Conditions of access and use can be found at

http://www.tandfonline.com/action/journalInformation?journalCode=imnc20
JOURNAL OF MICROENCAPSULATION, 2017
VOL. 34, NO. 1, 29–37
http://dx.doi.org/10.1080/02652048.2017.1280095

RESEARCH ARTICLE

Hot-melt extrusion microencapsulation of quercetin for taste-masking

Chia Miang Khor, Wai Kiong Ng, Parijat Kanaujia, Kok Ping Chan and Yuancai Dong
Division of Crystallisation & Particle Science, Institute of Chemical and Engineering Sciences, Jurong Island, Singapore

ABSTRACT ARTICLE HISTORY

Besides its poor dissolution rate, the bitterness of quercetin also poses a challenge for further develop- Received 13 September 2016
ment. Using carnauba wax, shellac or zein as the shell-forming excipient, this work aimed to microencap- Revised 9 December 2016
sulate quercetin by hot-melt extrusion for taste-masking. In comparison with non-encapsulated quercetin, Accepted 2 January 2017
the microencapsulated powders exhibited significantly reduced dissolution in the simulated salivary pH
6.8 medium indicative of their potentially good taste-masking efficiency in the order of zein > carnauba KEYWORDS
wax > shellac. In vitro bitterness analysis by electronic tongue confirmed the good taste-masking efficiency Quercetin; hot-melt
of the microencapsulated powders. In vitro digestion results showed that carnauba wax and shellac- extrusion; taste-masking;
microencapsulated powders presented comparable dissolution rate with the pure quercetin in pH 1.0 carnauba wax; shellac; zein
(gastric) and 6.8 (intestine) medium; while zein-microencapsulated powders exhibited a remarkably slower
dissolution rate. Crystallinity of quercetin was slightly reduced after microencapsulation while its chemical
structure remained unchanged. Hot-melt extrusion microencapsulation could thus be an attractive tech-
nique to produce taste-masked bioactive powders.

Introduction Microencapsulation can be achieved using various techniques,

Quercetin is a plant-derived flavonoid possessing excellent antioxi-
dant, anti-inflammatory and anti-cancer activities (Wach et al.,
2007; Kawabata et al., 2015). Its anti-obesity activity has recently
been proven although the virtual mechanisms have yet to be
explored (Ahn et al., 2008; Meydani and Hasan, 2010; Nabavi
et al., 2015; Wang et al., 2014). The poor solubility/dissolution rate
of quercetin, however, constitutes an obstacle for its pharmaceut-
ical and nutritional applications. A great deal of work has thus
been performed to overcome the dissolution issue (Sahoo et al.,
2011; Patel et al., 2012; Li et al., 2013; Gonçalves et al., 2015; Lai
et al., 2015). Its bitter taste, however, also constitutes another hur-
dle, since a low-consumer compliance would be expected from
the bitter compound-incorporated product (Drewnowski and
Gomez-Carneros, 2000). Hence, there have unmet needs to mask
the bitter taste of quercetin prior to its further downstream devel-
opment. Many approaches have been reported to mask or sup-
press the unpleasant taste of the bitter compounds. Examples
include the addition of flavours and sweeteners, microencapsula-
tion, inclusion with cyclodextrins, and ion exchange, which have
been comprehensively reviewed (Sun-Waterhouse and Wadhwa,
2013; Gaudette and Pickering, 2013; Maniruzzaman et al., 2014).
Among them, microencapsulation is a simple and attractive tech-
nique to produce taste-masked bioactive powders with relatively
less adverse impacts on the food matrix as well as consumers
(Champagne and Fustier, 2007; de Vos et al., 2010; Nazzaro et al.,
2012; Celli et al., 2015). Technically, microencapsulation is a pro-
cess which entraps, disperses or dissolves bioactives (core) within
a coating shell or a matrix. This leads to the generation of a phys-
ical barrier between the core agent and its surroundings.
Dissolution of the bitter bioactives in the oral cavity is
thereby retarded, leading to reduced interaction with the taste
buds. This helps to minimise the unpleasant taste perceived.
such as spray drying, fluidised bed coating, pan coating, coacerva-
tion, hot-melt extrusion and emulsification. Hot-melt extrusion is a
technique that offers many advantages over others, such as sim-
plicity, continuous operation, high throughput, and the ability to
operate in the absence of any organic solvent (Repka et al., 2008;
Maniruzzaman et al., 2012; Singh et al., 2013; Maniruzzaman et al.,
2014; Vynckier et al., 2014). During extrusion, bioactives (molten
or non-melted) are mixed with molten excipients that bind them
together to form granules, sheets or strands. Upon cooling and
solidification, the bioactives are enclosed by a coating shell or
buried within a matrix. A physical barrier between the core bioac-
tives and its surroundings is thus formed.
The purpose of this work is to utilise hot-melt extrusion to
microencapsulate quercetin powders for taste-masking. To our
knowledge, no work has been reported on the production of
taste-masked quercetin powders. Water-insoluble GRAS (Generally
Regarded As Safe) ingredients, including carnauba wax, shellac
and zein, were used as the coating shell-forming excipients.
Carnauba wax is obtained from the Brazilian palm tree, and is
mainly composed of esters. It possesses the highest melting point
(
85  C) among commercially available natural waxes. This attri-
bute makes it a good candidate for use as a microencapsulating
material (Milanovic et al., 2010; Kim et al., 2013; Madureira et al.,
2015). Shellac is a natural resin secreted by the insect Kerria lacca,
and comprises of polyhydroxy polycarboxylic esters and single
esters. It is a weak acid with an acid value of
80 and is not sol-
uble under acidic conditions (Farag and Leopold, 2011; Oehme
et al., 2011; Soradech et al., 2013). Zein is a water-insoluble prol-
amine isolated from corn gluten (Geraghty et al., 1981; Paliwal
and Palakurthi, 2014; Luo and Wang, 2014). These three GRAS
excipients, being water insoluble and resistant to water and mois-
ture penetration, have been broadly utilised as films, coatings, or

CONTACT Yuancai Dong dong_yuancai@ices.a-star.edu.sg Division of Crystallisation & Particle Science, Institute of Chemical and Engineering Sciences, Jurong
Island, Singapore
ß 2017 Informa UK Limited, trading as Taylor & Francis Group
30 C. M. KHOR ET AL.

micro/nano-particles for controlled release or to keep core active XDB-C18 column, 3.5 lm, 4.6  50 mm, Santa Clara, CA). Briefly, a
ingredient intact. In this work, microencapsulated powders were certain amount of the microencapsulated powders was first dis-
formulated by hot-melt extrusion of the excipients and quercetin solved in ethanol (for carnauba wax and shellac-microencapsu-
at specific elevated temperatures, followed by cooling and milling. lated powders) or 90% aqueous ethanol solution (for zein-
Dissolution of the microencapsulated powders in pH 6.8 medium microencapsulated powders) and then filtered through 450 nm
(enzyme-free saliva) within 1 min was performed to estimate their PTFE syringe filter. The drug concentration in the filtrate was
potential taste-masking efficiency (Shukla et al., 2009; Gittings measured using HPLC. The mobile phase, composed of 60%
et al., 2014). In vitro bitterness was further evaluated using methanol and 40% ultrapure water with 0.1% phosphoric acid,
TS-5000Z taste sensing system. Sample morphology and their was delivered at 1 mL/min. The temperature of the column was
physical states were investigated by field emission scanning elec- set to be 37  C and quercetin was detected at a UV absorbance of
tron microscopy (FESEM), X-ray diffraction (XRD) and differential 370 nm.
scanning calorimetry (DSC). The chemical structures of quercetin
before and after microencapsulation were examined by fourier-
transform infra-red spectroscopy (FTIR). Finally, in vitro digestion Dissolution in the simulated salivary pH 6.8 medium
of the microencapsulated powders was performed in pH 1.0 and
6.8 medium, simulating enzyme-free gastric and intestinal fluids, Dissolution of the microencapsulated quercetin powders was
respectively (Fernandes et al., 2002; Stippler et al., 2004; Lopes performed in pH 6.8 medium (simulating enzyme-free saliva)
et al., 2015). within 1 min (Shukla et al., 2009; Gittings et al., 2014). Briefly, a
certain amount of the microencapsulated powders containing
5 mg of quercetin was put into 2 mL pH 6.8 phosphate buffer
Materials and methods solutions with 0.1% (w/v) Tween 80 and stirred for 1 min at
Materials 200 rpm. An aliquot was then filtered through 450 nm PTFE fil-
ter. The concentration of the dissolved quercetin was immedi-
Carnauba wax was a gift from Strahl & Pitsch, Inc. (West Babylon, ately measured by HPLC as described above. The dissolution
NY). Shellac was provided by the Japan Shellac Industries, Ltd. was expressed as the percentage of the dissolved quercetin div-
(Osaka, Japan). Zein was kindly donated by Flo Chemical ided by the total quercetin added. The experiments were con-
Corporation (Ashburnham, MA). Quercetin, 37% hydrochloric acid, ducted in triplicate.
sodium phosphate dodecahydrate, potassium chloride, silver chlor-
ide, Tween 80, potassium phosphate monobasic, sodium phos-
phate dibasic and tartaric acid were obtained from Sigma-Aldrich Taste masking evaluation by TS-5000Z taste sensing system
(St. Louis, MO). Potassium hydroxide was supplied by Alfa Aesar
(Haverhill, MA). HPLC grade methanol and absolute ethanol was A certain amount of microencapsulated powders containing
from J.T. Baker (Center Valley, PA) and VWR Chemicals (Radnor, PA), 75 mg of quercetin was put into 30 mL simulated salivary pH 6.8
respectively. Ultrapure water was used throughout the work. medium and stirred at 200 rpm for 1 min. The resulting suspension
was immediately filtered through the 3 lm-pore size filter paper.
Bitterness of the filtered solution was evaluated using TS-5000Z
Preparation of microencapsulated quercetin powders
taste sensing system equipped with a C00 sensor and a reference
Microencapsulated quercetin powders were prepared by hot-melt electrode (Intelligent Sensor Technology Incorporation, Kanagawa,
extrusion technique. In brief, quercetin was mixed with the excipi- Japan). Each measurement lasted for 30 s. The measurement cycle
ent, i.e. carnauba wax, shellac or zein (plasticized with 20 w/w% comprised of a washing cycle, followed by measurement of the
water) for 30 min at different weight ratios in a turbular mixer standard reference solution (Vr). Next, the sample solution was
(Alphie powder mixer, Hexagon, Vadodara, India). Subsequently, measured and then lightly washed in standard reference solution
the physical mixtures were manually fed into the twin-screw melt before the “aftertaste” (Vs) was measured. Sensor output was cal-
extruder (Prism Eurolab 16, Thermo Scientific, Waltham, MA) pre- culated based on Vr (Woertz et al., 2011b). Interpolation difference
heated at 80  C for carnauba wax mixture, or 90  C for shellac and was performed to set the blank pH 6.8 medium as zero (no bitter-
zein mixtures. The extrusion was conducted continuously at a ness). Taste was expressed as bitterness response output. The ref-
screw rotation speed of 100 rpm without using a die. The extru- erence solution was made of 30 mM KCl and 0.3 mM tartaric acid
dates were collected, cooled to room temperature, and then
milled using Retsch MM 200 bead mill (Retsch GmbH, Haan, Table1. content and dissolution data in simulated salivary
Germany) for 8 min at 20 Hz in 25 ml stainless steel vessels with a pH 6.8 medium within 1 min (mean ± SD, n ¼ 3).
2.5 cm diameter stainless steel ball. The obtained microencapsu- Samples Content (%) Dissolution (%)
lated quercetin powders were then kept in screw-cap glass bottles Quercetin 100 3.14 ± 0.23
and placed in a dry cabinet at room temperature before further 80Q20C N.A. N.A.
analysis. The microencapsulated powders prepared from the mix- 70Q30C 65.47 ± 1.18 0.59 ± 0.14
tures, for example, 70% quercetin (Q) and 30% carnaubawax (C), 60Q40C 60.37 ± 0.74 0.56 ± 0.19
50Q50C 49.71 ± 0.25 0.52 ± 0.17
70% quercetin (Q) and 30% shellac (S) and 70% quercetin (Q) and 80Q20S N.A. N.A.
30% zein (Z) were denoted as 70Q30C, 70Q30S and 70Q30Z, 70Q30S 68.25 ± 0.82 4.12 ± 0.51
respectively. All other prepared samples were labelled in the same 60Q40S 59.46 ± 1.12 3.54 ± 0.09
way unless otherwise described. 50Q50S 49.46 ± 0.50 1.84 ± 0.05
40Q60S 39.91 ± 0.42 1.63 ± 0.08
80Q20Z 74.21 ± 0.46 0.40 ± 0.05
Quercetin content analysis 70Q30Z 63.10 ± 0.84 0.15 ± 0.04
60Q40Z 56.70 ± 1.05 0.21 ± 0.02
Quercetin content of the microencapsulated powders was ana- 50Q50Z 46.51 ± 0.38 0.21 ± 0.10
lysed using HPLC (Agilent HPLC 1100 with Agilent Zorbax Eclipse Melting not achieved.
 JOURNAL OF MICROENCAPSULATION 31

in ultrapure water. The cleaning solution was made of 0.1 M HCl Samples were mounted on aluminium stubs and sputtered with
in 30:70 ethanol:ultrapure water. The sensors were filled with 3.33 gold (Cressington 208HR, High Resolution Sputter Coater, Watford,
M KCl and 0.7 mM AgCl in ultrapure water. England) for 120 s at 20 mA before analysis. Scanning was per-
formed at 5.0 kV under lower secondary electron image (LEI) mode.
SEM
Morphologies of quercetin and the microencapsulated powders XRD
were examined by FESEM (JEOL JSM-6700 F, Tokyo, Japan). Diffractograms of quercetin and the microencapsulated powders
were investigated by D8-ADVANCE (BRUKER, Billerica, MA) X-ray
3.5 
diffractometer from 2–40 (2h) in steps of 0.017 using Cu Ka
radiation.
Bitterness sensory output (mv)

3.0 Quercetin
2.5
50Q50S
60Q40S DSC
2.0 70Q30S

1.5
40Q60S Thermograms of quercetin and the microencapsulated powders
50Q50C
60Q40C were examined using Diamond DSC Calorimeter (PerkinElmer,
1.0
Taste not perceived by the human tongue
70Q30C Waltham, MA). The samples were heated to 350  C at 10  C/min
50Q50Z under N2.
0.5 60Q40Z
Carnauba wax-microencapsulated powders
70Q30Z
0.0 Zein-microencapsulated powders 80Q20Z
FTIR
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0
% Mean quercetin dissolved
FTIR spectra were obtained in the range 400–4000 cm1 using FTS
3000MX FTIR (Bio Rad, Hercules, CA) KBr pellet method. 30 spec-
Figure 1. Bitterness sensory output of raw quercetin and microencapsulated
quercetin powders (sample denotation: for example, 50Q50C contains 50% quer- tral scans were performed for each sample with spectral width of
cetin and 50% carnauba wax; 50Q50S contains 50% quercetin and 50% shellac; 1 cm1, at a speed of 5 KHz.
50Q50Z contains 50% quercetin and 50% zein).

Figure 2. SEM images of (a) pure quercetin and microencapsulated powders: (b) 70Q30C (70% quercetin and 30% carnauba wax), (c) 40Q60S (40% quercetin and
60% shellac) and (d) 70Q30Z (70% quercetin and 30% zein).
32 C. M. KHOR ET AL.

(a) (b)

Shellac

Intensity
Intensity

40Q60S

Carnauba Wax

70Q30C
Quercetin

Quercetin
0 5 10 15 20 25 30 35 40
0 5 10 15 20 25 30 35 40
2θ
2θ

(c)

Zein
Intensity

70Q30Z

Quercetin

0 5 10 15 20 25 30 35 40
2θ
Figure 3. XRD patterns of raw querceitn and microencapsulated powders: (a) 70Q30C (70% quercetin and 30% carnauba wax), (b) 40Q60S (40% quercetin and 60%
shellac) and (c) 70Q30Z (70% quercetin and 30% zein).

In vitro digestion milling in this work. The yield of the process was around 70%
with some losses from the dead volume of the equipment
In vitro digestion profiles of quercetin and the microencapsulated
and adherence of some melts to the chamber wall and
powders were examined using USP dissolution apparatus, method II
extruder. The content of all the samples with different quer-
(Agilent 708 DS, Santa Clara, CA). In brief, to simulate enzyme-free
cetin/excipient ratio is exhibited in Table 1. With carnauba wax
gastric dissolution, 20 mg equivalent of quercetin or microencapsu-
and shellac as the excipients, production of 80Q20C and
lated powders were added to 750 mL of pH 1.0 medium containing
80Q20S was not successful, as the wax failed to melt at such
0.1% Tween 80 at 37  C with a stirring speed of 100 rpm. After
a high quercetin ratio. All other samples were successfully pre-
120 min, 250 mL of 0.2 M sodium phosphate solution with 0.1% (w/
pared. Compared with zein, the quercetin content of carnauba
v) Tween 80 preheated to 37  C was immediately added to adjust
wax and shellac-microencapsulated powders was closer to the
the pH of the medium to 6.8, simulating enzyme-free intestinal dis-
theoretical value, especially at lower quercetin content. This is
solution for another 120 min (Fernandes et al., 2002; Stippler et al.,
because carnauba wax and shellac is more easily melted than
2004; Lopes et al., 2015). Aliquots of 0.5 mL solution were with-
zein and possess good mixing with quercetin at lower quer-
drawn at specific time intervals and filtered through 450 nm PTFE
cetin content.
filters for HPLC analysis as described above. The experiments were
Since the taste-masking mechanism for microencapsulated
conducted in triplicate. Dissolution was expressed as a percentage
powders is their ability to inhibit the dissolution of unpleasant
of quercetin concentration detected at specific time intervals, div-
compounds in the oral cavity, in vitro dissolution in the simu-
ided by the total added quercetin concentration.
lated salivary pH 6.8 medium was measured as a convenient
and fast approach to estimate the potential taste-masking effi-
Results and discussion ciency (Shukla et al., 2009; Gittings et al., 2014). Table 1 exhib-
its the dissolution data of the microencapsulated quercetin
Dissolution in simulated salivary pH 6.8 medium
powders within 1 min in pH 6.8 medium. As shown,
Carnauba wax, shellac and zein-microencapsulated quercetin 3.14 ± 0.23% of the non-encapsulated pure quercetin was dis-
powders were prepared by hot-melt extrusion followed by solved within 1 min. Compared to non-encapsulated pure

(a) Wax
Endothermic
70Q30C
Pure Quercetin
0 50 100 150 200 250 300 350
Temperature (°C)
(c)
Endothermic
0 50 100
Temperature (°C)
Figure 4. DSC thermograms of microencapsulated powders: (a) 70Q30C (70% quercetin and 30% carnauba wax), (b) 40Q60S (40% quercetin and 60% shellac) and
(c) 70Q30Z (70% quercetin and 30% zein).
quercetin, the dissolution of 70Q30C dramatically decreased to
0.59 ± 0.14%. A reasonably good taste-masking efficacy can thus
be expected from carnauba wax-microencapsulated quercetin
powders, which is presumably due to the hydrophobicity and
€ using a variety of sensors, including umami, saltiness, sourness,
non-erodibility of the wax (Ozyazıcı et al., 2006; Raymond and
Champagne, 2014). A further decrease of quercetin ratio to pro-
duce 60Q40C and 50Q50C did not lead to significant changes
in dissolution. For shellac-microencapsulated quercetin, dissol-
ution of 70Q30S and 60Q40S gave rise to 4.12 ± 0.51% and
3.54 ± 0.09%, which was even higher than un-encapsulated quer-
cetin. This could be due to: (1) shellac having high solubility in
pH 6.8 medium, and thus failure to form an effective barrier
around quercetin to inhibit dissolution (Raymond and
Champagne, 2014); (2) the perfectness of the quercetin crystals
was diminished during the hot-melt extrusion process leading
to a faster dissolution rate. Further decreasing the quercetin
ratio to create 50Q50S resulted in a decrease in dissolution to
1.84 ± 0.05%, which plateaued off at 40Q60S with dissolution of
1.63 ± 0.0.08%. For zein, the dissolution of 80Q20Z was
0.40 ± 0.05%, which further reduced to 0.15 ± 0.04% for 70Q30Z.
Further decreasing of the quercetin ratio to produce 60Q40Z
and 50Q50Z did not yield a significant reduction in dissolution.
Comparing the three excipients used, zein exhibited the best
capacity to inhibit quercetin dissolution, which could be
explained by its high hydrophobicity and non-erosion in
the enzyme-free pH 6.8 medium (Hurtado-Lo pez and Murdan,
2002).
JOURNAL OF MICROENCAPSULATION 33
(b)
Shellac
Endothermic
40Q60S
Pure Quercetin
0 50 100 150 200 250 300 350
Temperature (°C)
Zein
70Q30Z
Pure Quercetin
150 200 250 300 350
In vitro bitterness evaluation by taste sensing system
Taste sensing system TS-5000Z, or electronic tongue, is capable of
detecting different tastes by measurement of electrical potential
sweetness, astringency and bitterness (Woertz et al., 2011a,
2011b). It is recommended by the manufacturer that bitterness
output values above one can be fully perceived by humans. Pure
shellac, zein and carnauba wax excipients were measured, and
had their bitterness response output values close to zero, indicat-
ing they are not bitter. Figure 1 exhibits the bitterness sensory
output vs. the mean percentage of quercetin dissolved. As can be
seen, the non-encapsulated quercetin had the highest bitterness
sensory output of 2.91 ± 0.08 mV, suggesting it is the most bitter,
compared with the microencapsulated powders. Shellac-
microencapsulated powders, i.e. 40Q60S, 50Q50S, 60Q40S and
70Q30S showed lower bitterness in comparison with the non-
encapsulated quercetin but significantly higher bitterness when
compared to carnauba wax and zein-microencapsulated powders.
This could be due to the higher percentage of quercetin dissolved
from shellac-microencapsulated powders. It is also noteworthy to
point out that, although the microencapsulated quercetin pow-
ders 60Q40S and 70Q30S had higher dissolution than the non-
encapsulated quercetin, their bitterness output was lower than
the latter. The reason is perhaps that, both shellac and quercetin
are weak acids; the shellac dissolved in pH 6.8 medium at high
concentration may thus interfere with the penetration of the
34 C. M. KHOR ET AL.

(a) (b) Shellac

Carnauba Wax

40Q60S

% Transmittance
% Transmittance

70Q30C

Quercetin
Quercetin

4000 3500 3000 2500 2000 1500 1000 500 4000 3500 3000 2500 2000 1500 1000 500
Wavenumber (cm−1) Wavenumber (cm−1)

(c)
Zein

70Q30Z
% Transmittance

Quercetin

4000 3500 3000 2500 2000 1500 1000 500

Wavenumber (cm−1)
Figure 5. FTIR of raw querceitn and microencapsulated powders: (a) 70Q30C (70% quercetin and 30% carnauba wax), (b) 40Q60S (40% quercetin and 60% shellac)
and (c) 70Q30Z (70% quercetin and 30% zein).

qucertin molecules into the sensor membrane (Tahara and XRD

Toko, 2013). The bitterness sensory output of all carnauba wax
(50Q50C, 60Q40C and 70Q30C, inside the circle in Figure 1)
and zein-microencapsulated powders (50Q50Z, 60Q40Z, 70Q30Z
and 80Q20Z, inside the oval in Figure 1) were less than 0.6 mv,
which is within the range that cannot be fully perceived by the
human tongue. The in vitro bitterness evaluation by taste sens-
ing system showed that carnauba wax and zein had potentially
better taste-masking efficiency than shellac. In addition, the bit-
terness sensory output results correlate well with the dissolution
data.
SEM
Figure 2 displays the typical SEM images of non-encapsulated
pure quercetin and the microencapsulated powders. Pure quer-
cetin can be seen to be mainly composed of approximately 10 lm
cuboid particles as well as a small fraction of 1–2 lm irregularly-
shaped fine particles. The size of the microencapsulated powders
ranged from dozens to more than 100 lm. This is because the
excipients (carnauba wax, shellac or zein) were in molten state
during the extrusion process (80–90  C), which bound the un-
melted quercetin (Tm
320  C) particles together leading to the
formation of the larger aggregates upon solidification and milling.
It is also found that carnauba wax-microencapsulated quercetin
powders turned out rather evenly distributed in size and rounded
in appearance, which is probably due to the crystalline nature of
the wax and thus ease of milling.
The XRD patterns of pure quercetin and its microencapsulated
powders are illustrated in Figure 3. Carnauba wax was observed
to exhibit two intense diffraction peaks suggesting its crystalline
nature, while shellac and zein displayed halo patterns indicative of
their amorphous nature. Pure quercetin presented its main intense
peaks at 2h of 10.74 , 12.36 , 13.98 , 26.42 and 27.18 , demon-
strating its crystallinity. All these characteristic diffraction peaks
can still be distinguished from the microencapsulated quercetin
powders, confirming that quercetin was still crystalline after
microencapsulation.
DSC
DSC thermograms of pure quercetin and microencapsulated
quercetin powders are given in Figure 4. Pure quercetin exhib-
ited a sharp and narrow melting endotherm at 320.5  C. For
carnauba wax-microencapsulated quercetin powders, two endo-
therms appeared at 84.1 and 316.5  C, corresponding to the
melting point of carnauba wax and quercetin respectively. The
melting point of quercetin after microencapsulation was
depressed by 4  C. In addition, the enthalpy decreased from
297 to 219 J/g, indicating reduced crystallinity of quercetin. The
thermogram of zein-microencapsulated quercetin powders
showed an endotherm at 136.7  C, which could be ascribed to
the dehydration of the water added to zein as a plasticiser

100
90
80
70 pH 1.0
Dissolution %
60
50
40
30
20
10
0
0 50 100
Time(min)
Figure 6. In vitro digestion profiles of raw quercetin and microencapsulated powders of 70Q30C (70% quercetin and 30% carnauba wax), 40Q60S (40% quercetin and
60% shellac) and 70Q30Z (70% quercetin and 30% zein).
during hot-melt extrusion. The melting endotherm of quercetin
completely disappeared in both zein and shellac-microencapsu-
lated powders. A possible explanation is that zein and shellac
decomposed prior to the melting point of quercetin leading to
masking effects (Lai et al., 2015).
FTIR
Figure 5 presents the FTIR spectra of pure quercetin and microen-
capsulated quercetin powders. Pure quercetin exhibited character-
istic bands at 1660 cm1 (C¼O stretching), 1600 cm1 (aromatic
ring stretch), 1200 cm1 and 1430 cm1 (OH phenolic stretch),
1310 cm1 and 1155 cm1 (C–O–C vibration), and finally the broad
peak from 3100 cm1 to 3500 cm1 (OH bond vibration) (Patel
et al., 2012; Li et al., 2013; Gonçalves et al., 2015; Lai et al., 2015).
All these characteristic bands were still observed in the
spectrums of microencapsulated quercetin powders, suggesting a
consistent chemical structure in quercetin before and after
microencapsulation.
In vitro digestion
Besides the good taste-masking efficiency, microencapsulated
powders should also possess the comparable dissolution rate in
comparison with the un-encapsulated quercetin to preserve its
bioavailability. Figure 6 presents the in vitro digestion (dissolution)
of the microencapsulated quercetin powders in pH 1.0
(0–120 min) and 6.8 (120–240 min) media, representing gastric and
intestinal fluids respectively. It is well known that the dissolution
of an agent from its microencapsulated form is governed by diffu-
sion, swelling or erosion of the coating layer/matrix (Ohara et al.,
2005). As can be seen, a burst dissolution to 29.8 ± 2.2% was
observed for pure quercetin within the first 15 min. After that,
quercetin continued to dissolve, but at a much slower rate, reach-
ing 35.8 ± 1.7% and 42.2 ± 1.4% dissolution within 120 min and
240 min respectively. The initial outburst is presumably contrib-
uted by fine quercetin particles of around 1–2 lm in size. The
larger specific surface area of fine quercetin particles resulted in a
faster dissolution rate. Carnauba wax-microencapsulated quercetin
powders displayed a linear dissolution pattern in pH 1.0 medium,
JOURNAL OF MICROENCAPSULATION 35
70Q30C
70Q30Z
40Q60S
Quercetin
pH 6.8
150 200 250
reaching 32.8 ± 0.6% within 120 min. Upon pH adjustment to 6.8,
dissolution slowed down and plateaued to 40.8 ± 2.4% after
240 min. The dissolution profile of carnauba wax-microencapsu-
lated powders can be ascribed mainly to its molecular diffusion,
as the wax was found to be non-swellable and non-erodible in
€
the dissolution medium (Ozyazıcı et al., 2006; Raymond and
Champagne, 2014). Shellac-microencapsulated quercetin powders
also exhibited a linear dissolution pattern in pH 1.0 medium, but
at a significantly slower rate: only 9.5 ± 0.7% of the encapsulated
quercetin was dissolved after 120 min. However, upon pH adjust-
ment from 1.0 to 6.8, dissolution abruptly increased to 38.1 ± 2.0%
(within 135 min) and 49.3 ± 1.4% (within 150 min); after that, dis-
solution reached a plateau. It has been reported that, shellac,
being a weak acid, is virtually insoluble in pH 1.0 medium but,
was soluble (erosion of 8.87% (w/w)) in pH 6.8 medium; in add-
ition, its swelling rate was pronouncedly increased from
8.6% in the acidic medium to more than 150% (w/w) in the pH
6.8 medium (Stummer et al. 2010; Limmatvapirat et al.
2004).Therefore, the dissolution of shellac-microencapsulated
quercetin powders in pH 1.0 medium is mainly governed by diffu-
sion, which brings about a slow and linear dissolution profile;
while swelling and erosion of shellac in pH 6.8 medium contrib-
uted to immense dissolution enhancement. Zein-microencapsu-
lated quercetin powders presented 8.1 ± 0.1% dissolution in
15 min; after that, it dissolved at an extremely slow rate with only
11.4 ± 0.4% and 11.3 ± 0.6% dissolution within 120 and 240 min,
respectively. As zein is highly hydrophobic and non-erodible in
the enzyme-lacking dissolution medium, quercetin dissolution
from the zein-microencapsulated powders was predominantly
controlled by diffusion leading to the much slower dissolution
rate (Hunt, 1951; Geraghty, et al. 1981; Zhang et al., 2015).
Conclusions
Carnauba wax, shellac and zein-microencapsulated quercetin pow-
ders were successfully prepared using the hot-melt extrusion tech-
nique. A significantly reduced dissolution rate of the carnauba
wax- and zein-microencapsulated powders compared to non-
encapsulated quercetin in the simulated salivary pH 6.8 medium
within 1 min suggests potentially good taste-masking efficiency.
36 C. M. KHOR ET AL.
In vitro bitterness evaluation by taste sensing system confirmed
their lower bitterness. Carnauba wax and shellac-microencapsu-
lated powders displayed comparable digestion profiles as that of
pure quercetin in pH 1.0 and 6.8 medium; while zein-microencap-
sulated powders presented a much slower dissolution rate. Based
on the taste-masking efficiency and in vitro digestion profiles, car-
nauba wax-microencapsulated quercetin is presumably suitable
for further downstream development (e.g. food formulation). Hot-
melt extrusion microencapsulation is thus an effective technology
to produce taste-masked bitter polyphenol compounds.
Acknowledgements
This work was supported by Agency for Science, Technology and
Research (ASTAR) of Singapore (project grant ICES/14–425A02).
Disclosure statement
The authors report no conflicts of interest. The authors alone are
responsible for the content and writing of this article.
Funding
This work was supported by Agency for Science, Technology and
Research (ASTAR) of Singapore (project grant ICES/14–425A02).
References
Ahn J, Lee H, Kim S, Park J, Ha T. The anti-obesity effect of quer-
cetin is mediated by the AMPK and MAPK signaling pathways.
Biochem Biophys Res Commun, 2008;373:545–9.
Celli GB, Ghanem A, Brooks MS-L. Bioactive encapsulated powders
for functional foods—a review of methods and current limita-
tions. Food Bioprocess Tech, 2015;8:1825–37.
Champagne CP, Fustier P. Microencapsulation for the improved
delivery of bioactive compounds into foods. Curr Opin
Biotechnol, 2007;18:184–90.
de Vos P, Faas MM, Spasojevic M, Sikkema J. Encapsulation for
preservation of functionality and targeted delivery of bioactive
food components. Int Diary J, 2010;20:292–302.
Drewnowski A, Gomez-Carneros C. Bitter taste, phytonutrients,
and the consumer: a review. Am J Clin Nutr, 2000;72:1424–35.
Farag Y, Leopold CS. Investigation of drug release from pellets
coated with different shellac types. Drug Dev Ind Pharm,
2011;37:193–200.
Fernandes CM, Teresa Vieira M, Veiga FJB. Physicochemical charac-
terization and in vitro dissolution behavior of nicardipine–cyclo-
dextrins inclusion compounds. Eur J Pharm Sci, 2002;15:79–88.
Gaudette NJ, Pickering GJ. Modifying bitterness in functional food
systems. Crit Rev Food Sci Nutr, 2013;53:464–81.
Geraghty D, Peifer MA, Rubenstein I, Messing J. The primary struc-
ture of a plant storage protein: zein. Nucleic Acids Res,
1981;9:5163–74.
Gittings S, Turnbull N, Roberts CJ, Gershkovich P. Dissolution
methodology for taste masked oral dosage forms. J Control
Release, 2014;173:32–42.
 Cocero
Gonçalves V, Rodrıguez-Rojo S, De Paz E, Mato C, Martın A,
M. Production of water soluble quercetin formulations by pres-
surized ethyl acetate-in-water emulsion technique using natural
origin surfactants. Food Hydrocoll, 2015;51:295–304.
Hunt J. The composition of gastric juice. J Physiol (Lond),
1951;113:419–24.
Hurtado-Lo pez P, Murdan S. Zein microspheres as drug/antigen
carriers: a study of their degradation and erosion, in the pres-
ence and absence of enzymes. J Microencapsul, 2002;23:
303–14.
Kawabata K, Mukai R, Ishisak A. Quercetin and related polyphe-
nols: new insights and implications for their bioactivity and bio-
availability. Food Funct, 2015;6:1399–417.
Kim IH, Lee H, Kim JE, Song KB, Lee YS, Chung DS, Min SC. Plum
coatings of lemongrass oil-incorporating carnauba wax-based
nanoemulsion. J Food Sci, 2013;78:E1551–9.
Lai F, Franceschini I, Corrias F, Sala MC, Cilurzo F, Sinico C, Pinib E.
Maltodextrin fast dissolving films for quercetin nanocrystal
delivery. A feasibility study. Carbohydr Polym, 2015;121:217–23.
Li B, Konecke S, Harich K, Wegiel L, Taylor LS, Edgar KJ. Solid dis-
persion of quercetin in cellulose derivative matrices influences
both solubility and stability. Carbohydr Polym, 2013;92:2033–40.
Limmatvapirat S, Limmatvapirat C, Luangtana-anan M, Nunthanid
J, Oguchi T, Tozuka Y, Yamamoto K, Puttipipatkhachorn S.
Modification of physicochemical and mechanical properties of
shellac by partial hydrolysis. Int J Pharm, 2004;278:41–9.
Lopes MA, Abrahim-Vieira B, Oliveira C, Fonte P, Souza AM, Lira T,
Sequeira JA, Rodrigues CR, Cabral LM, Sarmento B, et al.
Probing insulin bioactivity in oral nanoparticles produced by
ultrasonication-assisted emulsification/internal gelation. Int J
Nanomed, 2015;10:5865–80.
Luo Y, Wang Q. Zein-based micro-and nano-particles for drug and
nutrient delivery: a review. J Appl Polym Sci, 2014;131:40696.
Madureira AR, Campos DA, Fonte P, Nunes S, Reis F, Gomes AM,
Sarmento B, Pintado MM. Characterization of solid lipid nano-
particles produced with carnauba wax for rosmarinic acid oral
delivery. RSC Adv, 2015;5:22665–73.
Maniruzzaman M, Boateng JS, Bonnefille M, Aranyos A, Mitchell
JC, Douroumis D. Taste masking of paracetamol by hot-melt
extrusion: an in vitro and in vivo evaluation. Eur J Pharm
Biopharm, 2012;80:433–42.
Maniruzzaman M, Boateng JS, Chowdhry BZ, Snowden MJ,
Douroumis D. A review on the taste masking of bitter APIs: hot-
melt extrusion (HME) evaluation. Drug Dev Ind Pharm,
2014;40:145–56.
Meydani M, Hasan ST. Dietary polyphenols and obesity. Nutrients,
2010;2:737–51.
Milanovic J, Manojlovic V, Levic S, Rajic N, Nedovic V, Bugarski B.
Microencapsulation of flavors in carnauba wax. Sensors (Basel),
2010;10:901–12.
Nabavi SF, Russo GL, Daglia M, Nabavi SM. Role of quercetin as an
alternative for obesity treatment: you are what you eat! Food
Chem, 2015;179:305–10.
Nazzaro F, Orlando P, Fratianni F, Coppola R. Microencapsulation
in food science and biotechnology. Curr Opin Biotecnol,
2012;23:182–6.
Oehme A, Valotis A, Krammer G, Zimmermann I, Schreier P.
Preparation and characterization of shellac-coated anthocyanin
pectin beads as dietary colonic delivery system. Mol Nutr Food
Res, 2011;55:S75–S85.
Ohara T, Kitamura S, Kitagawa T, Terada K. Dissolution mechanism
of poorly water-soluble drug from extended release solid dis-
persion system with ethylcellulose and hydroxypropylmethylcel-
lulose. Int J Pharm, 2005;302:95–102.
€
Ozyazıcı M, Go €kçe EH, Ertan G. Release and diffusional modeling
of metronidazole lipid matrices. Eur J Pharm Biopharm, 2006;
63:331–9.
Paliwal R, Palakurthi S. Zein in controlled drug delivery and tissue
engineering. J Control Release, 2014;189:108–22.

Patel AR, Heussen PC, Hazekamp J, Drost E, Velikov KP. Quercetin
loaded biopolymeric colloidal particles prepared by simultan-
eous precipitation of quercetin with hydrophobic protein in
aqueous medium. Food Chem, 2012;133:423–9.
Raymond Y, Champagne CP. Dissolution of lipid-based matrices in
simulated gastrointestinal solutions to evaluate their potential
for the encapsulation of bioactive ingredients for foods. Int J
Food Sci, 2014;2014:749630.
Repka MA, Majumdar S, Kumar Battu S, Srirangam R, Upadhye SB.
Applications of hot-melt extrusion for drug delivery. Expert
Opin Drug Deliv, 2008;5:1357–76.
Sahoo NG, Kakran M, Shaal LA, Li L, M€ uller RH, Pal M, Tan LP.
Preparation and characterization of quercetin nanocrystals.
J Pharm Sci, 2011;100:2379–90.
Shukla D, Chakraborty S, Singh S, Mishra B. Fabrication and
evaluation of taste masked resinate of risperidone and its
orally disintegrating tablets. Chem Pharm Bull, 2009;
57:337–45.
Singh A, Majumdar S, Deng W, Mohammed N, Chittiboyina A,
Raman V, Shah S, Repka MA. Development and characterization
of taste masked Efavirenz pellets utilizing hot melt extrusion.
J Drug Del Sci Technol, 2013;23:157–63.
Soradech S, Limatvapirat S, Luangtana-anan M. Stability enhance-
ment of shellac by formation of composite film: Effect of gelatin
and plasticizers. J Food Eng, 2013;116:572–80.
Stippler E, Kopp S, Dressman J. Comparison of US pharmacopeia
simulated intestinal fluid TS (without pancreatin) and phos-
phate standard buffer pH 6.8, TS of the international
JOURNAL OF MICROENCAPSULATION 37
pharmacopoeia with respect to their use in in vitro dissolution
testing. Dissolut Technol, 2004;11:6–11.
Stummer S, Salar-Behzadi S, Unger FM, Oelzant S, Penning M,
Viernstein H. Application of shellac for the development of pro-
biotic formulations. Food Res Int, 2010;43:1312–20.
Sun-Waterhouse D, Wadhwa SS. Industry-relevant approaches for
minimising the bitterness of bioactive compounds in functional
foods: a review. Food Biopro Technol, 2013;6:607–27.
Tahara Y, Toko K. Electronic tongues – a review. IEEE Sens J,
2013;13:3001–11.
Vynckier AK, Dierickx L, Voorspoels J, Gonnissen Y, Remon JP,
Vervaet C. Hot-melt co-extrusion: requirements, challenges and
opportunities for pharmaceutical applications. J Pharm
Pharmacol, 2014;66:167–79.
Wach A, Pyrzyn ska K, Biesaga M. Quercetin content in some food
and herbal samples. Food Chem, 2007;100:699–704.
Wang S, Moustaid-Moussa N, Chen L, Mo H, Shastri A, Su R, Bapat
P, Kwun I, Shen CL. Novel insights of dietary polyphenols and
obesity. J Nutr Biochem, 2014;25:1–18.
Woertz K, Tissen C, Kleinebudde P, Breitkreutz J. A comparative
study on two electronic tongues for pharmaceutical formulation
development. J Pharm Biomed Anal, 2011a;55:272–81.
Woertz K, Tissen C, Kleinebudde P, Breitkreutz J. Taste sensing sys-
tems (electronic tongues) for pharmaceutical applications. Int J
Pharm, 2011b;417:256–71.
Zhang Y, Cui L, Che X, Zhang H, Shi N, Li C, Chen Y, Kong W. Zein-
based films and their usage for controlled delivery: origin, classes
and current landscape. J Control Release, 2015;206:206–19.