Food Hydrocolloids 43 (2015) 540e546

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Preparation and characterization of nanoemulsion encapsulating

curcumin
T.P. Sari a, Bimlesh Mann a, *, Rajesh Kumar a, R.R.B. Singh b, Rajan Sharma a,
Minaxi Bhardwaj a, S. Athira a
a
Dairy Chemistry Division, National Dairy Research Institute, Karnal, Haryana 132001, India
b
Dairy Technology Division, National Dairy Research Institute, Karnal 132001, India

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

Article history: Curcumin is the most active and least stable bioactive component of turmeric (Curcuma longa) plant. In
Received 7 March 2014 the present study, an attempt has been made to overcome the instability during processing and
Accepted 11 July 2014 bioavailability problems of curcumin by nanoencapsulation technology and the effect was evaluated by
Available online 22 July 2014
simulated digestion study. Curcumin was encapsulated in medium chain triglyceride oil droplets of
nanoemulsion prepared by ultrasonification using whey protein concentrate-70 and Tween-80 as
Keywords:
emulsifiers with an encapsulation efficiency of 90.56 ± 0.47%. The prepared nanoemulsion has particles of
Curcumin
average diameter 141.6 ± 15.4 nm and zeta potential of
6.9 ± 0.2 mV. In vitro release kinetics of cur-
Nanoencapsulation
Bioavailability
cumin from nanoemulsion by simulated gastrointestinal studies showed that the curcumin nanoemulsion
Nanoemulsion was relatively resistant to pepsin digestion but pancreatin causes release of curcumin from nanoemulsion.
Simulated gastrointestinal digestion The slow release of curcumin from the nanoemulsion was supposed to increase bioavailability. The total
antioxidant activity of curcumin nanoemulsion was reduced from 3.53 ± 0.11 mM Trolox/mg of curcumin
to 3.33 ± 0.02 mM Trolox/mg of curcumin after encapsulation. The prepared nanoemulsion was stable to
pasteurization, different ionic strengths (0.1e1 M) and pH ranging from 3.0 to 7.0. The study has
important implication in the formation and design of encapsulated bioactive systems.
© 2014 Elsevier Ltd. All rights reserved.

1. Introduction To improve the bioavailability of curcumin, numerous ap-

Curcumin, the bioactive component obtained by the extraction
and purification of ground rhizomes of Curcuma longa has been
found to exert wide range of beneficial biological and pharmaco-
logical activities including antioxidant (Sharma, 1976), anti-
inflammatory (Chainani-Wu, 2003), antimicrobial (De et al.,
2009) and anticancer (Aggarwal, Kumar, & Bharti, 2003) in clin-
ical trials. But studies addressing the metabolism and uptake of
curcumin had shown that either no curcumin or little amount was
detected in serum or tissue after administration (Dhillon et al.,
2008). The main reasons attributing to the low bioavailability are
supposed to be the poor solubility of curcumin in aqueous media,
rapid hydrolysis followed by molecular fragmentation at physio-
logical pH and inactivity of its metabolic products (Lin, Pan, & Lin-
Shiau, 2000).
* Corresponding author.
E-mail addresses: bimleshmann@gmail.com, bmann@rediffmail.com (B. Mann).
proaches have been undertaken by many researchers including,
preparation of nanocurcumin (Bisht et al., 2007), liposomal curcu-
min (Li, Braiteh, & Kurzrock, 2005), curcuminephospholipid com-
plex (Liu, Lou, Zhao, & Fan, 2006), and chelation with metals (John,
Kuttan, & Krishnankutty, 2002). Micro/nanoencapsulation tech-
nology of poorly water soluble bioactives has attracted wide
attention in the food and pharmaceutical industry for the past few
years for various applications like protection of bioactivity and their
controlled release for improving bioavailability. The encapsulation
of polyphenols overcome the drawbacks of their instability, alle-
viate unpleasant tastes or flavors, as well as improve the half-life of
the compound in vivo and in vitro (Augustin & Hemar, 2009;
Mozafari et al., 2008). Nanoencapsulation is generally employed
to improve solubility, stability and bioactivity of various oil-soluble
phytochemicals due to their small droplet size and high kinetic
stability. Among the nanometric encapsulation systems, nano-
emulsions are particularly suitable and widely accepted for food
applications (McClements, Decker, & Weiss, 2007). Encapsulation
into nanoemulsion-based delivery systems of bioactive compounds

http://dx.doi.org/10.1016/j.foodhyd.2014.07.011
0268-005X/© 2014 Elsevier Ltd. All rights reserved.
 T.P. Sari et al. / Food Hydrocolloids 43 (2015) 540e546
characterized by low solubility in aqueous phase represents an
effective approach to improve the dispersion of the bioactives into
food products, to protect them against degradation or interaction
with other ingredients, to reduce the impact on organoleptic
properties of the food and to improve their bioavailability (Donsi,
Sessa, Mediounic, Mgaidic, & Ferrari, 2011). A kinetically stable
suspension can be obtained by adding stabilizers, emulsifiers or
texture modifiers in the emulsion systems.
The slow release of curcumin from nanoparticulate curcumin
formulation increased the bioavailability of delivered curcumin.
Unlike native curcumin, curcumin was detected in serum for a long
duration time period as observed after 24 h of its administration
in vivo. Whereas, in native case, the curcumin level was subse-
quently decreased in serum with time and not detectable after 1 h,
indicating rapid metabolism of native curcumin in physiological pH
(Mohanty & Sahoo, 2010). Wang et al. (2008) prepared nano-
emulsion of curcumin and found that the anti-inflammatory ac-
tivity improved than the conventional emulsion. Bioavailability of
curcumin was improved by encapsulation with polylactic-co-
glycolic acid nanoparticles (Anand et al., 2010).
Emulsion based colloidal delivery systems are commercially
used in food, pharmaceutical and cosmetics industries. So, it is
necessary to establish their stability to environmental stress and its
effect on the physico-chemical characteristics. It has been reported
that the physical and chemical stability of nanoemulsions greatly
dependent on storage temperature, pH, and ionic strength (Qian,
Decker, Xiao, & McClements, 2012; Rao & McClements, 2011).
There is a need to optimize the right composition of nanoemulsions
such as decrease creaming, flocculation and coalescence for
different commercial applications.
In order to design functional foods with potential to promote
human health, it is necessary to examine the digestibility and
release of food components under simulated gastrointestinal (GI)
conditions (Shani-Levi, Levi-Tal, & Lesmes, 2013). The delivery
systems based on nanoencapsulation have been designed to control
the targeted release of encapsulated material at a specific location
in the human GI tract. The emulsion containing medium chain
triglyceride (MCT) as carrier lipid has been reported to substantially
increase the bioaccessibility of curcumin (Ahmed, Li, McClements,
& Xiao, 2012). Knowledge of digestion kinetics of foods in human
GI tract is also required in establishing processing conditions at the
manufacturing stage to achieve desirable release of nutrients. In the
present study, we focused on the preparation, physico-chemical
characterization and in vitro digestion kinetics of curcumin
nanoemulsion.
2. Materials and methods
2.1. Materials
Curcumin (95% purity) was procured from Plant Lipids Pvt. Ltd.
(Kerala, India) and used without further treatment. MCT-60 with
predominantly MCT C8(58.6%) & C10(41.4%) was obtained from
Kamani Oil Industries Pvt. Ltd. (Mumbai, India). Tween-80 was
Fig. 1. Methodology of preparation of curcumin encapsulated nanoemulsion.
541
purchased from MERCK (Merck Specialities Pvt. Ltd., Worli, Mum-
bai). Whey protein concentrate (WPC-70) was procured from
Modern Dairy Pvt. Ltd. (Karnal, India). Pepsin and pancreatin were
purchased from Sisco Research Laboratories Pvt. Ltd. and Hi Media
laboratories Pvt. Ltd., Mumbai, India, respectively.
2.2. Preparation and characterization of curcumin nanoemulsion
Oil-in-water nanoemulsion was prepared in two stages using
the method described by Jafari, He, and Bhandari (2007) with
modification. Curcumin (10e40 mg) was added to 100 ml of total
emulsion along with MCT-60 (0.5e2%), surfactant Tween-80
(2e10% w/w) and emulsifier WPC-70 (0e1% w/w) in Millipore
water. The coarse emulsion was then prepared using magnetic
stirrer under ambient temperature for different time intervals. Fine
emulsion was prepared by sonifying the coarse emulsion by opti-
mizing the sonification time using ultrasonicator (Fig. 1).
Emulsion stability was measured by centrifugation at 1300  g
(Kubota, Tokyo, Japan) at 5  C for 30 min and heating at 80  C for
30 min. The stable sample was further analyzed for physico-
chemical characterization. Particle size distribution was deter-
mined by dynamic laser light scattering method using Malvern
Zetasizer Nano ZS90 (Malvern Instruments, UK). Zeta potential, the
electrical charge on the oil droplets in the emulsions was determined
under holder temperature of 25  C and electrical voltage 3.9 V.
2.3. Determination of encapsulation efficiency
The encapsulation efficiency was determined and calculated by
method of Surassmo, Min, Bejrapha, and Cho (2010) with modifica-
tion. Nanoemulsion was passed through Vivaspin concentrators of
MWCO (Molecular Weight Cut Off) 100 kDa and centrifuged at
1300  g and 5  C for 30 min. The permeate was collected to calculate
encapsulation efficiency by measuring its total phenolic content.
Total phenolic content of nanoemulsions and permeate was analyzed
by FolineCiocalteu's method given by Zheng and Wang (2001).
2.4. Determination of stability of nanoemulsion under processing
conditions
The effect of different processing conditions like heating (63  C
for 30 min and 95  C for 10 min), pH (3e7) and ionic strength
(0.1e1 M NaCl) was studied which will be applicable for their
commercial utilization. The particle size distribution, zeta potential
and stability were measured after exposing the nanoemulsions to
each treatment.
2.5. Antioxidant activity of curcumin nanoemulsion
Radical scavenging activity of the curcumin emulsions before and
after encapsulation was determined by DPPH (2, 2 diphenyl-1-picryl
hydrazyl) method given by Williams, Cuvelier, and Berset (1995).
The results were expressed as trolox equivalent antioxidant capacity
(TEAC) values i.e. mM of trolox equivalent/mg of curcumin.
542 T.P. Sari et al. / Food Hydrocolloids 43 (2015) 540e546
2.6. Release kinetics of curcumin from nanoemulsion
The release mechanism of encapsulated curcumin under simu-
lated GI conditions was evaluated by mimicking the physiological
situation in the upper tract (stomach and small intestine) of human
body. Simulated gastric fluid (SGF) was prepared by the method
given by Sanchez, Fernandez-Garcia, Margolles, Reyes-Gavilan, and
Ruas-Madiedo (2010). Curcumin nanoemulsion (adjusted to pH 1.5)
was mixed with SGF in the ratio of 1:3 in a series of test tubes. The
mixture was placed in a shaking water bath at 37  C for 2 h which
represents the average duration of GI transit. The entire samples
were subdivided in different test tubes with equal volume. During
the course of experiment the samples were removed after every
hour for testing the percentage of curcumin released with respect
to time. The release of curcumin from nanoemulsion was evaluated
after deactivation of enzyme by keeping the emulsions at 90  C/
10 min. Simulated intestinal fluid (SIF) was prepared by the method
given by Liang et al. (2012) with slight modifications. The SIF con-
tained 4 mg/ml pancreatin and 25 mg/ml bile salts. The pH was
adjusted to 7.5 using 1 N NaOH. The sample after 2 h of gastric
digestion was mixed with SIF and allowed to incubate for further
3 h at 37  C. Samples were removed from the water bath as
mentioned above after each hour and release of curcumin from
nanoemulsion was measured as the total phenolic contents after
deactivating of enzymes.
2.7. Statistical analysis
All statistical analysis was done using MS-EXCEL-2010 package.
All measurements were done for at least three replicates. Results
are presented as means ± standard error of means (SEM).
3. Results and discussion
3.1. Optimization and characterization of curcumin nanoemulsion
Different concentration of both core (curcumin and oil) and
coating materials (emulsifiers, and/or co-surfactant) at different
mass ratios were used to optimize the stable formulations (Fig. 1).
The conditions for the preparation of nanoemulsions were opti-
mized and stable nanoemulsions were obtained by sonification at
4  C for 15 min. The most stable formulation was obtained with
40 mg curcumin dissolved in a suitable solvent (2% MCT) þ 2%
Tween-80 þ 0.5% WPC-70 having particle size of 141.6 ± 15.4 nm,
zeta potential of
6.9 ± 0.2 mV and poly dispersity index (PDI) of
0.273. The pH of the nanoemulsion was 5.57 ± 0.1 and it was stable
for 27 ± 2 days at 25  C (Table 1). The relatively lower value of PDI
for nanoemulsion can be correlated with more stability on storage.
The reason for the aggregation of nanoemulsion during storage in
globular protein stabilized emulsions is flocculation characterized
by the exposure of non-polar groups in the protein (Kim, Decker, &
McClements, 2002). In the present study, both WPC and Tween-80
Table 1
Physico-chemical characteristics of curcumin nanoemulsion.
Nanoemulsion composition 2% MCT* þ 2% Tween-80 þ 0.5%
WPC-70** þ 40 mg curcumin
Size (nm) 141.6 ± 15.4
Zeta potential (mV)
6.9 ± 0.2
Poly dispersity index 0.273
Stability at room temperature (days) 27 ± 2
pH 5.57 ± 0.1
Encapsulation efficiency (%) 90.56 ± 0.47
MCT*: medium chain triglycerides and WPC-70**: whey protein concentrates-70.
Values mentioned above is mean ± SEM (n ¼ 3).
were tried alone and in combination to get stable formulations.
Preliminary studies in our laboratory (Sari et al., 2013) have indi-
cated that individual use of both Tween-80 (1e10%) and WPC-70
(1e5%) did not show a positive result with respect to their stabil-
ity as studied during heating followed by centrifugation.
Researchers have successfully prepared emulsions of curcumin
with relatively higher concentration of Tween-80 (Nakagawa,
Sowasod, Charinpanitkul, Soottitantawat, & Tanthapanichakoon,
2011; Wang et al., 2008). There are two types of emulsifiers being
used in foods i.e. low molecular weight (e.g. Tween-80) and high
molecular weight (e.g. proteins) surfactants. At similar bulk con-
centrations (W/V), low molecular weight surfactants decrease the
surface or interfacial tension to a greater extent than the macro-
molecular surfactants. This difference mainly related to differences
in orientation and configuration of these surfactants at the inter-
face. In the case of low molecular mass surfactants, the entire
molecule adsorbs and instantaneously orients itself and this par-
titioning of the entire molecule between the two phases facilitates
a maximum reduction of interfacial tension. Although proteins are
less surface active than low molecular mass surfactants, but
emulsions formed by low molecular mass surfactants are mostly
unstable. This is because proteins, in addition to lowering interfa-
cial tension, can form a continuous viscoelastic membrane like film
around oil droplets via noncovalent intermolecular interactions
and via covalent disulphide cross-linking whereas low molecular
mass surfactants cannot form such a viscoelastic films. Thus,
emulsions which contain both low molecular and macromolecular
surfactants, the stability of colloidally dispersed phases is primarily
dependent on protein films adsorbed at the interfaces (Damodaran,
1997). Moreover, in case of water soluble non-ionic surfactant,
whether the addition occurs before or after emulsification has
relatively little influence on the competitive adsorption behavior
(Courthaudon, Dickinison, Matsumura, & Williams, 1991). In light
of the above discussion we may say that in the present formulation
both are present at the interface.
The zeta potential roughly characterizes the surface charge of
the emulsion particles. High absolute values lead to repulsive forces
between particles, which may improve the physical stability of
multiphase systems (Muller, 1996). The zeta potential of the
emulsion has been presented in Table 1. The relatively low value of
zeta potential in the prepared nanoemulsion may be due to the
presence of relatively higher amount of Tween-80 used in formu-
lations so that the surface charge is contributed only by whey
proteins. Electrostatic and steric repulsion are the two main sta-
bilizing forces for whey protein stabilized emulsions. Electrostatic
interactions between similarly charged droplets are repulsive, and
may be either short range or long range depending on ionic
strength (Demetriades, Coupland, & McClements, 1997). When
droplets come so close together that the adsorbed emulsifier layer
begins to overlap, some short-range interactions become impor-
tant, such as steric, thermal fluctuation and hydration forces
(Israelachvili, 1992). So it indicates the emulsion droplets stabilized
by protein due to electrostatic and steric repulsion but electrostatic
repulsion predominates due to the surface charge on the proteins.
Whereas the oil droplets of emulsions are stabilized by Tween-80
against aggregation by steric (rather than electrostatic) repulsion
due to relatively large hydrophilic (polyoxyethylene) head groups
of adsorbed Tween molecules (Klang & Valenta, 2011).
3.2. Encapsulation efficiency of curcumin in its nanoemulsion
The encapsulation efficiency of curcumin into nanoemulsion
was 90.56 ± 0.47% (Table 1) which was evaluated by considering
total phenolic content as a marker. To determine the encapsulation
efficiency the samples (native and nanoencapsulated) were passed
 T.P. Sari et al. / Food Hydrocolloids 43 (2015) 540e546 543

Table 2
Effect of pH, ionic strength and heating on particle size and zeta potential of cur-
cumin nanoemulsions.

Particle Zeta
size (nm) potential (mV)

Effect of pH
Control nanoemulsion 141.6 ± 15.4
6.9 ± 0.2
(pH: 5.57 ± 0.1)
pH 3.0 163.4 ± 7.03 þ2.03 ± 0.1
pH 5.0 143 ± 8.08
6.9 ± 0.2
pH 7.0 140.5 ± 5.74
6.26 ± 0.4

Effect of ionic strength

0.1 M NaCl 137.9 ± 11.7
1.83 ± 0.1
0.5 M NaCl 140.3 ± 9.2
1.25 ± 0.3
1.0 M NaCl 140.7 ± 7.4
0.523 ± 0.1

Effect of heating
63  C/30 min (pasteurization) 139.0 ± 11.7
4.19 ± 0.7
95  C/10 min (boiling) 531.9 ± 6.76
1.65 ± 0.2

Values mentioned above is mean ± SEM (n ¼ 3).

the conformation of the b-lactoglobulin monomer. These changes

Fig. 2. Images of curcumin solution (15 ml) passed through 100 kDa MWCO mem-
brane; a) native curcumin dissolved in ethyl alcohol, b) nanoencapsulated curcumin
(2% oil þ 2% Tween-80 þ 0.5% whey protein concentrate-70).
through 100 kDa MWCO membrane. From Fig. 2, it can be depicted
that native curcumin dissolved in ethyl alcohol passed through the
membrane while, nanoencapsulated curcumin retains in the
retentate.
3.3. Changes in size and zeta potential of nanoemulsion under
different processing conditions
3.3.1. Effect of heating
The influence of different heat treatment on the particle size
distribution of the curcumin nanoemulsion was evaluated at 63  C
for 30 min (pasteurization) and 95  C for 10 min (boiling) and re-
sults are presented in Fig. 3 and Table 2. The average size of the
particles changed slightly after pasteurization while boiling causes
gradual increase in the average size due to aggregation (Fig. 3). The
nanoemulsion was unstable to aggregation of the particles at
elevated temperature most likely due to the denaturation of whey
protein which causes aggregation. On heating to 70  C, the dimers
of b-lactoglobulin dissociate to form monomers (Hambling,
McAlpine, & Sawyer, 1992) followed by some critical change in
lead to the exposure of hydrophobic groups and the free sulphydryl
group (Cys121) (Relkin, Meylheuc, Launay, & Raynal, 1998), resulting
in a reactive monomer which can propagate an aggregation reac-
tion leading to the formation of non-native dimers, trimmers, tet-
ramers and larger aggregates by polymerization. b-Lactoglobulin
being the most abundant whey protein, dominates the overall
behavior of whey protein products. Qian et al. (2012) investigated
the influence of temperature on the physical and chemical stability
of b-carotene enriched nanoemulsions and concluded that the
nanoemulsions were prone to droplet aggregation at elevated
temperatures (>37  C).
The zeta potential of the nanoemulsion was also measured as a
function of heat treatments. The denaturation of whey protein also
caused the change in the surface charge of particles. The zeta po-
tential was
6.9 ± 0.2 mV (Table 1) initially and the magnitude of
the charge decreased appreciably with increasing temperature.
After pasteurization and boiling, the particles had relatively low net
charges, with the magnitude of the zeta potential being
4.19 ± 0.7
and
1.65 ± 0.2 mV (Table 2), respectively.
3.3.2. Effect of pH
The particle size and zeta potential of the nanoemulsion were
evaluated after adjusting to different pH (2.0e7.0) (Table 2 and
Fig. 4). Average particle size increased with decrease in acidity of
the emulsion i.e. from 163.4 to 140.5 nm at pH 7.0. The higher mean

Fig. 3. Average particle size and particle size distributions of curcumin nanoemulsion Fig. 4. Average particle size and particle size distributions of curcumin nanoemulsion
after pasteurization (63  C/30 min) and boiling (95  C/10 min). maintained at different pH (3.0, 5.0 and 7.0).
544 T.P. Sari et al. / Food Hydrocolloids 43 (2015) 540e546
value of particle size at low pH was may be due to the aggregation
of particles at pH values close to the isoelectric point of whey
protein. This can also be correlated with the reduction in the net
negative charge at low pH values which leads to particle aggrega-
tion and corresponding increase in the particle size.
There was a significant difference in the zeta potential of the
nanoemulsion with change in pH from 3.0 to 7.0. The particles at pH
3.0 have net positive charge (þ2.03 ± 0.1 mV) compared to particles
at pH 7.0 (
6.26 ± 0.4 mV) (Table 2). This is due to the fact that the
surface charge of the milk proteins shifts to positive at pH below
the isoelectric point. This shift in the zeta potential indicates that
the surface charge of the prepared nanoemulsion is contributed by
milk proteins. Rao and McClements (2011) studied the impact of
nano/microemulsions on various environmental stress and found
that relatively stable nanoemulsions were formed at pH 6.0 and 7.0,
but extensive particle growth/aggregation occurred at lower and
higher pH values, which was attributed to either chemical (hy-
drolysis) or physical (electrical charge) effects. The reduction in the
electrical charge on the particles at lower pH value would reduce
the electrostatic repulsion between them, thereby leading to ag-
gregation (Rao & McClements, 2012).
3.3.3. Effect of ionic strength
Salts are common food additives and are also present in the
human gastrointestinal tract. Salts may therefore affect the func-
tional performance of colloidal delivery systems in a product or
after ingestion. For these reasons, the influence of salt (0.1e1 M
NaCl) on properties of nanoemulsion was examined. The addition
of salt did not cause any significant change in the particle size
(Fig. 5). The zeta potential of the nanoemulsion shifted toward zero
due to the decrease in the electrostatic repulsion between the
molecules upon addition of salt and consequently the charge on the
molecules was reduced (Table 2). Thus addition of salt may lead to
destabilization of the nanoemulsions. The destabilization of the
nanoemulsions at high salt concentrations can be attributed to
screening of the electrostatic repulsion between the protein coated
droplets by the salt ions. At relatively low salt levels, the electro-
static repulsion is still sufficiently strong to overcome the weak van
der Waals and hydrophobic attraction, but above a critical salt level,
it is no longer strong enough so that the attractive forces dominate,
leading to droplet aggregation (McClements, 2005).
3.4. DPPH scavenging activity measurements of curcumin
nanoemulsion
The antioxidant activity of the encapsulated curcumin is slightly
lower than that of unencapsulated curcumin. The total antioxidant
Fig. 5. Average particle size and particle size distributions of curcumin nanoemulsion
maintained at different ionic strengths (0.1e1.0 M NaCl).
activity of curcumin nanoemulsion determined by DPPH radical
scavenging assay was 3.33 ± 0.02 mM Trolox/mg of curcumin,
whereas radical scavenging activity of native curcumin was
3.53 ± 0.11 02 mM Trolox/mg of curcumin. To exert the bioactivity,
antioxidants have to be bio-accessible, i.e., released from the food
matrix and solubilized (Bouayed, Hoffmann, & Bohn, 2011).
Although the antioxidant activity of the encapsulated compound is
lower than unencapsulated, the fact is that, the encapsulation not
only protects curcumin from degradation but may also preserve the
antioxidant activity.
Donsi et al. (2011) evaluated the effect of the delivery systems of
curcumin by comparing the antioxidant activity of the encapsu-
lated compound with that of unencapsulated curcumin. The anti-
oxidant activity of curcumin encapsulated in solid lipid
nanoemulsion using FRAP assay was 0.996 ± 0.07 mM L-ascorbic
acid equivalent while that of unencapsulated curcumin shows a
value of 2.504 ± 0.06 mM L-ascorbic acid equivalent.
3.5. In vitro digestion of curcumin nanoemulsion
The digestion of encapsulated bioactives in the gastrointestinal
tract is a complex process and its impact on the release of bioactive
component plays a major role in the uptake, distribution as well as
bioavailability of the component. The gastric and intestinal digest
was analyzed for % release of total phenolic content (curcumin)
from nanoemulsion (Fig. 6). The results indicated that the encap-
sulated curcumin was very stable and not released from the de-
livery system by the action of pepsin. During simulated gastric
digestion (2 h), over 90% of the encapsulated curcumin was
retained in emulsion. The resistance of nanoemulsion
toward pepsin digestion may be attributed to the resistance of b-
lactoglobulin against pepsin digestion. Since WPC-70 is composed
of 50e60% b-lactoglobulin and native b-lactoglobulin has been
reported to be almost resistant to breakdown in the gastric
compartment following simulated digestion (Fu, Abbott, & Hatzos,
2002; Schmidt, Meijer, Slangen, & Van Beresteijn, 1995) due to its
compact globular structure (Guo, Fox, & Flynn, 1995), 8.48% release
of curcumin after gastric digestion in the present study may be due
to highly acidic condition.
The synergistic interactions between enzymes and acidic-
alkaline pH conditions result in a significant breakdown of food
Fig. 6. Release of curcumin from the nanoemulsion under simulated gastro-intestinal
digestion condition by using simulated gastric fluid (SGF) and simulated intestinal fluid
(SIF).
 T.P. Sari et al. / Food Hydrocolloids 43 (2015) 540e546
materials including emulsions (Sarkar, Goh, Singh, & Singh, 2009).
Stability of emulsion droplets in the gastric environment is a
desirable attribute for an encapsulation system in order to protect
the encapsulant from the harsh gastric environment (Anal & Singh,
2007; Vandenberg, Drolet, Scott, & De la Noue, 2001). But in case of
intestinal digestion, the incubation with SIF resulted in destabili-
zation of the emulsion and approximately 77% of the encapsulated
curcumin was released within 2 h of incubation (Fig. 6). Fu et al.
(2002) and Schmidt et al. (1995) reported that b-lactoglobulin is
almost completely digested by pancreatic enzymes. This indicates
that the prepared nanoemulsions of curcumin are stable to gastric
enzymes but can be destabilized with subsequent release of cur-
cumin in presence of pancreatin. Bossios et al. (2011) reported that
only b-lactoglobulin was broken down to smaller peptides after
gastro-duodenal digestion although a sizable amount of intact
protein still remained. Simulated intestinal fluid (SIF) is a mixture
of pancreatin and bile salts. Bile salts may change the interface
which facilitates activity of lipase present in pancreatin and helps in
the release of curcumin. Similar observation was made by Sarkar,
Horne, and Singh (2010) that bile salt preferentially adsorbs at
the oilewater interface displacing the protein or binding on to the
protein coated droplets, thereby allowing lipase to act. In the
presence of bile salts, coalescence between emulsion droplets
seemed to occur, resulting in oiling-off and breakdown of the
b-lactoglobulin stabilized emulsion system. This was probably due
to the lipolytic activities of pancreatin, being favored by the bile
salts displacing the interfacial b-lactoglobulin. Apart from
the possible lipolytic action of the pancreatin causing coalescence,
the proteolysis of the adsorbed protein layer (due to the tryp-
sinechymotrypsin fractions present in pancreatin) at the droplets
surface of the emulsion would possibly have resulted in reduced
surface viscosity compared to intact interface and facilitates
coalescence.
Benzariaa, Maresca, Taiebc, and Dumay (2013) reported that
enzymatic digestion of PC (phosphor casein) curcumin complexes
was quite resistant to pepsin treatment in simulated stomach
digestion. Pancreatin treatment caused greater and faster release of
curcumin, showing 38e77% of total curcumin released within
5e60 min incubation, and 81% after 3 h incubation. Incubation in
simulated intestinal environment resulted in destabilization of the
emulsion and approximately 60% of the encapsulated curcumin
was released within 2 h of incubation (Tikekar, Pan, & Nitin, 2013).
Devadasu, Wadsworth, and Kumar (2012) reported that curcumin
is mainly found in the intestinal tract after oral administration of
nanoparticles. Our study also supported this statement as majority
of curcumin got released with intestinal enzymatic action.
4. Conclusion
Curcumin being highly unstable and hydrophobic is difficult to
incorporate in aqueous food systems. For this reason the curcumin
was encapsulated inside carrier oil in an emulsion form. The results
in the present study showed that curcumin releases slowly from
nanoemulsion under simulated digestion. These results have
important implication in the design of nanoemulsions of curcumin
for commercial applications. It is concluded that the nano-
encapsulation of highly lipophilic and unstable compounds is an
effective platform to increase the hydrophilicity, bioaccessibility
and to protect them from degradation.
Acknowledgments
We are grateful for the financial support given by National Dairy
Research Institute, Karnal, Haryana and NAIP, ICAR for this work.
(Grant No. C30029), We would like to thank Plant Lipid Pvt. Ltd.,
545
Kerala (India) and Modern Dairy Pvt. Ltd., Haryana (India) for
providing curcumin and WPC-70, respectively, for the emulsion
preparation.
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