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Synthesis and study the properties of StNPs/gum nanoparticles for salvianolic
acid B-oral delivery system
Article in Food Chemistry · February 2017
DOI: 10.1016/j.foodchem.2017.02.059
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2. Synthesis and study the properties of StNPs/gum nanoparticles for
salvianolic acid B-oral delivery system
Xiaojing Li, Shengju Ge, Jie Yang, Ranran Chang, Caifeng Liang, Liu Xiong, Mei Zhao, Man Li, Qingjie Sun ⇑
College of Food Science and Engineering, Qingdao Agricultural University, Qingdao, Shandong Province 266109, China
a r t i c l e i n f o
Article history:
Received 19 September 2016
Received in revised form 26 January 2017
Accepted 13 February 2017
Available online 16 February 2017
Keywords:
Polyphenol delivery
Ternary system
Stability
Short-chain glucan
Controlled release
a b s t r a c t
To fabricate stable sized and shaped controlled release delivery systems for salvianolic acid B (Sal B), dif-
ferent food gums were individually added to short-chain glucan solution to prepare starch nanoparticles
(StNPs)/gum nanocomposites by self-assembly, and Sal B was embedded in situ. The results showed that
size of StNPs was reduced to ca. 45 nm with the addition of chitosan and rosin, which decreased by over
50% than that of StNPs without the gum. The StNPs/guar gum nanocomposites had the largest size
(109.2 nm) among samples of StNPs with gums. The StNPs with chitosan and gum arabic exhibited an
obvious core-shell structure. The loading capacities of Sal B in StNPs, StNPs/chitosan, and StNPs/gum ara-
bic nanocomposites were 5.2, 8.26 and 8.08%, respectively. The in vitro release of Sal B from StNPs/gum
nanocomposites were sustained and prolonged for over 12 h, indicating that StNPs/gum nanocomposites
are good candidates to control Sal B release.
Ó 2017 Elsevier Ltd. All rights reserved.
1. Introduction
Salvianolic acid B (Sal B) is the largest quantity of water-soluble
bioactive polyphenol of salviae miltiorrhizae (Hu, Liang, Luo, Zhao,
& Jiang, 2005). It is considered to be one of natural functional plant
metabolite with strongest antioxidant capacity currently known
(Zhao et al., 2008). Many reports showed that Sal B could be used
to scavenge free radicals and resist oxidant, and perform better
than vitamin E, vitamin C and mannitol (Zhou, Xie, Xu, Liang, &
Wei, 2014). In addition, Sal B as dietary supplement is usually
added to enhance the functionality of food (Xia et al., 2014; Lim,
Lee, Kim, Shin, & Kwon, 2016).
Current pharmacokinetic reports elucidated the low oral
bioavailability of Sal B. The oral bioavailability of salvianolic acid
B in freely moving rats was calculated to be 2.3% by Wu et al.
(2006). Moreover, Gao, Han, Zhang, Fang, and Wang (2009)
reported that the oral bioavailability of Sal B in dogs was calculated
to be only 1.07 ± 0.43%, which was too low to ameliorate blood vis-
cosity in beagle dogs. Recently, Zhu and Zhang (2015) studied the
pharmacokinetics of Sal B monomer in normal and hyperlipidemic
rats and concluded that Sal B could not play a therapeutic role
according to statistical results due to their low bioavailability.
Nano-sized particles have been used as the encapsulating mate-
rial in active ingredients or drug delivery carriers to decrease insta-
bility and improve bioavailability (D’Addio & Prud’homme, 2011).
Nanoparticles with particle size of less than 200 nm could be
absorbed by small intestinal epithelium cell, and thus enhance
the absorption of encapsulated phenolic phytochemicals (Li,
Jiang, Xu, & Gu, 2015). In addition, the longevity and stability of
the encapsulated drug increased, and its side effects reduced
(Craparo, Bondì, Pitarresi, & Cavallaro, 2011).
Starch, as the most abundantly available and low-cost biomate-
rial, has been widely used for synthesizing starch-based nanopar-
ticles for various biomedical and industry applications (Han,
Borjihan, Bai, Chen, & Jing, 2008). Previously, our group found that
short-chain glucan debranched from waxy maize starch could form
starch nanoparticles (StNPs) via recrystallization (Sun, Li, Dai, Ji, &
Xiong, 2014). However, the poor dispersibility of the synthesized
StNPs in aqueous solutions presents huge limitations. Surfactants
can be adsorb on the surface of nanoparticles and significantly
improve their dispersion stability in aqueous media (Raclesa,
Iacoba, Butnaruc, Sacarescua, & Cazacua, 2014). Food polysaccha-
rides are also natural surfactants for stabilizing nanoparticles as
opposed to conventional surfactants. Many studies have reported
evidence describing the effects of food gums on stabilizing
nanoparticles. Gum arabic could be used to fabricate stable sodium
caseinate nanoparticles resistant to high temperature and strong
acid and alkaline condition (Ye, Edwards, Gilliland, Jameson, &
Singh, 2012), and could improve the dispersion stability of zein
http://dx.doi.org/10.1016/j.foodchem.2017.02.059
0308-8146/Ó 2017 Elsevier Ltd. All rights reserved.
⇑ Corresponding author at: Changcheng Road, Chengyang District, Qingdao,
China.
E-mail address: phdsun@163.com (Q. Sun).
Food Chemistry 229 (2017) 111–119
Contents lists available at ScienceDirect
Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

3. nanoparticles at pH 3.0–8.0 (Chen & Zhong, 2015). In addition,
selenium nanoparticles could keep favorable dispersion stability
in gum arabic solutions for about 30 days (Kong et al., 2014). More-
over, Hu and McClements (2015) reported that core-shell zein-
alginate nanoparticles had smaller size and were more stable over
a range of temperatures, pH values, and salt concentration than
pure zein nanoparticles.
On the other hand, food polysaccharides could also be used as a
template to prepare size-controlled and uniform nanoparticles or
nanofibers. Cellulose and chitosan (CS) can interact with metal
nanoparticles to control the nucleation of nanoparticles (Wei,
Sun, Qian, Ye, & Ma, 2009). Lokanathan, Ahsan Uddin, Rojas, and
Laine (2014) found that cellulose nanocrystals were capable of
minimizing the growth of silver nanoparticles (AgNP) through
steric hindrance. Yu, Si, Chen, Bian, and Chen (2006) reported that
sodium alginate (SA) played a template-like role in the synthesis of
polyaniline-SA nanofibers and affected the average diameter by
changing the concentration of SA.
To the best of our knowledge, there has not been a relevant
study on the monodispersed and size-controlled StNPs prepared
by adding polysaccharides as a template. The objectives of this
study were to (1) utilize different polysaccharides to coat or be
used as a template to prepare stable StNPs; (2) prepare stable
ternary nanoparticles using Sal B, StNPs, and different food gums
for the treatment of cerebrovascular diseases; (3) understand
interaction the forces between StNPs and different polysaccha-
rides; and (4) determine the stability of complex nanoparticles
and the in vitro controlled release of Sal B. This research will bring
some enlightenment for the potential of starch nanocarrier’s appli-
cating in food and medicinal field.
2. Materials and methods
2.1. Materials
Waxy maize starch (98% amylopectin) was purchased from
Tianjin Tingfung Starch Development Co., Ltd (Tianjin, China).
Pullulanase (E.C.3.2.1.41, 6000 ASPU/g, 1.15 g/mL) was supplied
by Novozymes Investment Co. Ltd. (Bagsvaerd, Denmark). Pancre-
atin (batch No. SLBC2100V) was purchased from Sigma-Aldrich
Chemical Co. (St. Louis, MO, USA). Salvianolic acid B (Sal B, 95%)
was purchased from Shanghai Moqi Biotechnology Co., Ltd. (Shang-
hai, China). Chitosan (CS, degree of deacetylation 84.5%) was pur-
chased from Qingdao Haipu Biotechnology Co., Ltd. (Qingdao,
China). Guar gum (GG) was purchased from Qingdao Tianxin Food
Additives Co., Ltd. (Qingdao, Shandong, China). Gum arabic (GA),
sodium alginate (SA), and rosin (RS) were obtained from Tianjin
Kaixin Chemical Industrial Co., Ltd. (Tianjin, China). All other
reagents were of analytical grade.
2.2. Preparation of gum solution
The stock solution of GG, GA, SA, CS, and RS were prepared by
dissolving 0.1 g gum in 10 ml distilled water (1% acetic acid
solution for CS, absolute ethyl alcohol for RS), respectively, and
was stirred overnight at room temperature. The solution was
centrifuged at 2000g for 5 min to remove the insoluble materials.
2.3. Synthesis of StNPs/gum nanocomposites
According to the method of Sun et al. (2014), short-chain glucan
powder was prepared and then dissolved in deionized water (10%,
w/v) by heating in a sealed tube at 120 °C for 30 min. After cooling
down to 25 °C, different food gum solutions were added into the
short-chain glucan solution to reach 0.1% (w/v) with vigorous
stirring, or equal distilled water as the substitute of food gum solu-
tion was added to prepare native StNPs. For the CS sample, to make
it soluble in the SGC solution, 1% (w/v) acetic acid was added. Then
the solutions were stored at 25 °C for 12 h. The suspensions were
washed several times with distilled water until neutrality was
achieved and were then vacuum freeze-dried to obtain StNPs/
gum nanocomposites or StNPs powder.
2.4. Synthesis of Sal B loaded StNPs/gum nanocomposites
Sal B (2 g) was added into the SGC solution (100 mL), and then
different food gums were added to reach 0.1% (w/v), with vigorous
stirring. The solutions were then stored at 25 °C for 12 h, and sus-
pensions were washed and vacuum freeze-dried as section 2.3. The
solution obtained from washing the Sal B loaded StNPs/gum
nanocomposites each time was mixed together for determining
loading capacity and encapsulation efficiency.
2.5. Determination of morphologies of the nanoparticles
Morphologies of StNPs, StNPs/gum nanocomposites, and StNPs/
gum-Sal B nanocomposites were taken with a Hitachi (Tokyo,
Japan) 7650 transmission electron microscope with an acceleration
voltage of 80 kV. The treatment condition of samples was in accor-
dance with our previous report (Li et al., 2016).
2.6. Determination of average size and size distribution of the
nanoparticles
The average size and size distribution of the nanoparticles were
determined by dynamic light scattering using a Malvern Zetasizer
Nano (Malvern Instruments Ltd., UK) equipped with a He-Ne laser
(0.4 mW, 633 nm) and a temperature-controlled cell holder. The
measurements were performed according to the method of Liu,
Zhao, Ren, Zhao, and Yang (2011). The mean intensity-weighted
diameter was recorded.
2.7. Determination of zeta potential of the nanoparticles
The StNPs and StNPs/gum nanocomposites suspensions (0.01%,
w/v) were measured for their electrophoretic mobility by laser
Doppler velocimetry using a Malvern Zetasizer Nano, following
the method reported by Li et al. (2016).
2.8. Particle stability against environmental stresses
2.8.1. pH stability
According to the method of Joye, Nelis, and McClements (2015),
the StNPs and StNPs/gum nanocomposites were dispersed in dis-
tilled water at 0.1% (w/v), and their pH was adjusted at room tem-
perature by adding NaOH or HCl (0.05 mM) to obtain pH values
ranging from pH 3.0 to 9.0. The samples were then stored for
0.5 h, and their average size and zeta potential were measured as
described above (sections 2.7 and 2.8).
2.8.2. Temperature stability
The StNPs and StNPs/gum nanocomposites were dispersed in
distilled water at 0.1% (w/v) and incubated in water baths set at
different temperatures (30–90 °C) for 30 min. The samples were
then cool down to room temperature and then stored for 0.5 h,
and their average size and zeta potential were analyzed.
2.8.3. Ionic strength
The StNPs and StNPs/gum nanocomposites were dispersed in
NaCl solutions (0, 100, 200, 300, 400, and 500 mM) at 0.1% (w/v).
112 X. Li et al. / Food Chemistry 229 (2017) 111–119

4. The samples were then stored for 0.5 h at room temperature for the
analysis of the particle size distribution and zeta potential.
2.9. Differential scanning calorimeter (DSC)
The thermal properties of StNPs and StNPs/gum nanocomposite
samples were investigated using a differential scanning calorime-
ter (DSC1, Mettler-Toledo, Schwerzenbach, Switzerland), as
described by Sun et al. (2014).
2.10. X-ray diffraction pattern (XRD)
The X-ray diffraction pattern of the StNPs and StNPs/gum
nanocomposite samples was studied with an X-ray diffractometer
(Bruker AXS Model D8 Discover) under the conditions described by
Watcharatewinkul, Puttanlek, Rungsardthong, and Uttapap (2009).
Before determination, the samples were equilibrated to 20% mois-
ture content in a saturated relative humidity chamber for 24 h at
room temperature. The scanning range and rate were 5–40° (2h)
and 1.0°/min, respectively.
2.11. Fourier transform infrared (FTIR) spectra
The infrared spectra of StNPs and StNPs/gum nanocomposite
samples were recorded on an FTIR spectrophotometer (NEXUS-
870, ThermoNicolet Corporation), as described by Kunal, Banthia,
and Majumdar (2008). All samples were collected using KBr
method, and were then subjected to attenuated total reflectance
(ATR) spectroscopy in the range of 4000–400 cm1
, and the resolu-
tion was 4 cm1
.
2.12. Loading capacity and encapsulating efficiency
The mixed solution obtained from washing the Sal B loaded
StNPs nanocomposites (Section 2.4) was diluted to a suitable con-
centration, and the amount of Sal B was quantified at a wavelength
of 286 nm using a Persee UV-1810 spectrophotometer against a
predetermined Sal B standard calibration curve. The loading capac-
ity (LC) and encapsulating efficiency (EE) of the Sal B was calcu-
lated as follows:
EE ð%Þ ¼
Total amount of Salvianolic acid B weight Free Salvianolic acid B weight
Total amount of Salvianolic acid B weight
100
LC ð%Þ ¼
Total amount of Salvianolic acid B weight Free Salvianolic acid B weight
Total amount of nanoparticle weight
100
2.13. In vitro release studies
An in vitro drug release from the Sal B loaded StNPs nanocom-
posites and Sal B loaded StNPs/gum nanocomposites was mea-
sured using a United States Pharmacopeia XXIII dissolution
apparatus 2 (paddle apparatus) according to Liu et al. (2015) with
minor modifications. Briefly, nanocomposites powder (1 g) was
dispersed in dissolution media (900 mL) at a paddle rotation speed
of 50 rpm at 37 °C. The drug release analysis was continuously per-
formed in different dissolution media. In the first stage, drug
release was measured in simulated gastric fluid (SGF, pH 1.2,
0.1 M HCl solution with 0.05 M NaCl, 3.2 g/L pepsin) for 2 h. Then,
the pH of the medium was adjusted to 6.8 by adding an appropri-
ate amount of anhydrous Na3PO4. Pretreated pancreatin (0.45 g)
was then added to the medium to simulate intestinal environment,
and the drug release was measured in simulated intestinal fluid for
4 h. Afterwards, the pH was increased to 7.4 by adding anhydrous
Na3PO4 to simulate colon fluid, and this stage was maintained for
up to 24 h. Samples were withdrawn at 0, 1, 2, 4, 6, 8, 12, 18 and
24 h, and equal fresh media were added at regular intervals. The
amount of Sal B released from the nanoparticles was determined
by a Persee UV-1810 spectrophotometer.
2.14. Statistical analysis
Each measurement was carried out using at least three fresh,
independently prepared samples. The data were subjected to sta-
tistical analysis using SPSS 17.0 (SPSS Inc., Chicago, United States),
analyzed using analysis of variance (ANOVA), and expressed as
mean values ± standard deviations. Differences were considered
at a significant level of 95% (p 0.05).
3. Results and discussion
3.1. Morphology and size of StNPs/gum nanocomposites
The food gum-templated StNPs’ surface morphology and actual
particle size were determined by transmission electron microscopy
(TEM) and DLS. Fig. 1A shows that the StNPs without food gums
were spherical in shape and 70–90 nm in size. When food gums
were added to short-chain glucan solution to serve as template
material, four kinds of StNPs in spherical, spindly, core-shell, and
irregular shapes were prepared. Their size could be divided into
two groups: one of ultra-fine size with an average diameter of 9–
30 nm and the other larger with average diameter of 50–150 nm.
The obtained nanocomposites were much smaller in size compared
to conventional colloidal delivery systems. For example, liposomes
prepared by phosphatidylcholine were close to spherical with lar-
ger diameters (200–250 nm) (Lin et al., 2014). Moreover, Isacchi
et al. (2011) reduced the size of liposomes (140 nm) by increasing
the liposomal surface charge with the method of chemical grafting.
The homogeneous StNPs/CS nanocomposites (Fig. 1B) were
spherical, and their average diameter was in the range of 20–
50 nm. CS chains could be adsorbed on the surface of nucleus
formed during the recrystallization of the short-chain glucan and
limit further deposition of short-chain glucan, leading to the for-
mation of nanoparticles with reduced size. What is new is that
the StNPs/CS nanocomposites showed a clear core-shell structure
with some cavities, similar to the structure of vesicles. The
StNPs/GA nanocomposites (Fig. 1D) had morphology similar to that
of the StNPs/CS nanocomposites. The hollow nanoparticles had
diameters of about 50 nm. The vesicle-like structures are helpful
for the embedding of active ingredients. This is probably the first
report of vesicle self-assembled by short-chain glucan and CS
(GA). Generally, CS and GA coated nanoparticles exhibited solid
shape with a larger mean size compared to neat nanoparticles
(Chen Zhong, 2015; Wang, Yang, Yuan, Gao, Huang, 2016; Ye
et al., 2012). The StNPs/SA nanocomposites were spindly, monodis-
perse, and homogeneous (Fig. 1E). Yu et al. (2006) reported that SA
could play a significant role in the synthesis of polyaniline-SA
nanofibers, because SA with a linear structure could act as ‘‘nano-
fiber seeding” to control the shape of polyaniline nanomaterial.
When GG was added to short-chain glucan solution to fabricate
nanoparticles, irregular nanoparticles with obvious aggregation
could be observed (Fig. 1C), GG could be strongly adsorbed onto
the surface of the nucleus, and its complicated side chains
(Fig. S1) created huge steric hindrance. The RS-templated StNPs
showed a small particle size with an irregular spherical shape
(Fig. 1F). In addition, there were many ultrafine nanoparticles on
the surface of the irregular nanoparticles. This could be due to
the growth orientation of the short-chain glucan on the surface
of RS molecule.
X. Li et al. / Food Chemistry 229 (2017) 111–119 113

5. 3.2. Stability of StNPs/gum nanocomposites
3.2.1. pH stability
As shown in Fig. 2, the zeta potential of all the suspensions had
a similar decreased trend when the pH changed from 3.0 to 10.0.
Nanoparticles size was closely related with their zeta potential
(Dai et al., 2015). The high absolute values of zeta potential could
increase electrostatic repulsion among nanoparticles to prevent
aggregation. The zeta potentials of StNPs/GA nanocomposites’
and StNPs/SA nanocomposites’ suspensions went from 5.37 mV
to 20.2 mV and from 10.5 mV to 24.6 mV as pH values
increased from 3.0 to 10.0, respectively, which corresponded to
their decreased mean diameters. Similarly, Chen and Zhong
(2015) reported that zeta potential of GA coated zein nanoparticles
exhibited a decreasing magnitude trend when the pH increased
from 3.0 to 8.0, which induced their decrease in mean diameters.
The RS-templated StNPs were stable in size (43.4–46.9 nm) across
the entire pH range (3.0–10.0) and were ideal for the encapsulation
and transportation of nutraceuticals and drugs.
3.2.2. Temperature stability
The zeta potential of StNPs with or without food gums was
stable at temperatures from 30 °C to 60 °C (Fig. S2), which could
be attributed to the maintenance of the StNPs’ integrity. With
increasing heating temperature (80–90 °C), above the melting tem-
perature (see Table 1), the integrity of the StNPs and nanocompos-
ites was destroyed, resulting in the decreased surface electrical
charge of the nanoparticles.
Heating treatment could increase the collision frequency
between nanoparticles, and StNPs structure was destroyed when
exceeding their melting temperature, which may promote particle
aggregation. As shown in Fig. S2, all StNPs, with or without food
Fig. 1. TEM images and particle size distribution of starch nanoparticles without (A) or with chitosan (B), Guar gum (C), Gum arabic (D), Sodium alginate (E) and Rosin (F).
114 X. Li et al. / Food Chemistry 229 (2017) 111–119

6. gums, were stable in particle size at temperatures from 30 °C to
60 °C. The diameter of the StNPs, StNPs/CS nanocomposites, and
StNPs/GG nanocomposites began to increase at 70 °C, which could
have a close correlation with their low melting temperature (see
Table 1). In contrast, the StNPs/SA nanocomposites, StNPs/GA
nanocomposites, and StNPs/RS nanocomposites were destabilized
when temperature increased to 80 °C. When the temperature
was 90 °C, the size of the StNPs increased to 634.5 nm, which is
almost 7 times larger than that of the StNPs at 30 °C. The increase
in the size of the StNPs with food gums was less than that of the
StNPs after exposure to 90 °C; particularly, the StNPs/SA nanocom-
posites and StNPs/RS nanocomposites increased only to 155.7 nm
and 123.4 nm. This could be attributed to the interaction between
SA or RS and short-chain glucan. Similarly, Joye, Davidov-Pardo,
and McClements (2015) reported that sodium caseinate could
make zein nanoparticles keep stable at 90 °C, though the size of
zein nanoparticles increased from 100 nm to 270 nm due to the
coating of sodium caseinate.
3.2.3. Ionic strength stability
Salt plays an important role in improving flavor and extending
the shelf life of food products during food processing. Therefore,
research on the effect of salt concentration on the nanoparticles
stability is meaningful. The stability of StNPs and StNPs/gum
nanocomposites against ionic strength is shown in Fig. S3. All
StNPs sample suspensions were stable in particle size with the
addition of salt (200 mM NaCl). However, the particle size of
StNPs increased with increasing salt concentration (200 mM
NaCl). This could be attributed to the decreasing net charge of
the StNPs, from 10.39 mV to 4.4 mV with salt concentrations from
200 mM to 500 mM (Fig. S3A). Similarly, the dispersion of GA
coated zein nanoparticles was stable in salt solution (300 mM
NaCl) (Chen Zhong, 2015). Compared with pure StNPs, GA- and
SA-templated StNPs were much more stable (Fig. S3B) within the
range of salt concentration tested, which could be due to stronger
electrostatic repulsion provided by GA and SA. Recently, Joye et al.
(2015) found that the coating of anionic pectin could make gliadin
nanoparticles more stable at 50–200 mM NaCl. The coating of food
gums on the surface of nanoparticles could increase steric repul-
sion among nanoparticles. This was why StNPs/RS nanocomposites
were stable in size at the high salt concentration condition, though
their charge was lower than that of the StNPs without food gums.
3.3. Differential scanning calorimeter
The melting temperatures and enthalpies change (DH) of differ-
ent food gum-templated StNPs were determined, and the results
are shown in Table 1. Onset, peak, and conclusion temperatures
(To, Tp, and Tc) of StNPs were about 65 °C, 88 °C, and 101 °C, respec-
tively. These parameters were lower than those of StNPs prepared
by nanoprecipitation with a 1:4 ratio of short-chain glucan to abso-
lute ethanol (81 °C, 95 °C, 109 °C, respectively) (Qiu et al., 2016).
The incorporation of food gums markedly increased the To, Tp,
and DH of the StNPs (p 0.05). Interactions of polysaccharides
and starch might reduce starch-chain mobility to influence gela-
tinization properties of starch (Xu et al., 2012). Therefore, melting
temperature of nanocomposites increased when polysaccharide
interacted with short-chain glucan. Tp increased while Tc-To
decreased, indicating that food gums did indeed favor formation
of StNPs with uniform crystallinity and fine crystalline. Nanoparti-
cles with a larger number of double-helices and a more ordered
crystalline array would have higher DH (Altay Gunasekaran,
2006). As shown in Table 1, the DH of the StNPs exhibited signifi-
cant increase with the addition of food gums (p 0.05), which
could be that short-chain glucan assembled on the surface of food
gums (as a template) and formed more ordered crystalline arrays.
-30
-25
-20
-15
-10
-5
0
5
10
15
20
3 4 5 6 7 8 9 10
Zeta
Potential
(mV)
pH
0
20
40
60
80
100
120
140
3 4 5 6 7 8 9 10
D
(nm)
pH
Fig. 2. Zeta potential and average particle diameter of starch nanoparticles without
(r) or with chitosan (j), Guar gum (N), Gum arabic ( ), sodium alginate ( ) and
rosin (d) under different pH.
Table 1
Thermal characteristics and crystallinity degree of starch nanoparticles (StNPs) without or with different food gums.
Sample Onset Temperature/°C Peak
Temperature/°C
Conclusion Temperature/°C DH/Jg1
Crystallinity Degree (%)
StNPs 65.64 ± 1.04d
88.99 ± 0.55d
101.93 ± 0.67b
11.20 ± 0.12f
53.72 ± 0.98c
StNPs/CS 73.96 ± 0.51a
92.46 ± 0.73b
103.99 ± 0.43a
15.05 ± 0.03e
58.16 ± 1.69b
StNPs/GG 70.43 ± 0.32c
90.86 ± 0.44c
104.22 ± 0.55a
18.87 ± 0.15a
62.35 ± 1.47a
StNPs/GA 72.15 ± 0.78b
91.09 ± 0.77b
103.57 ± 0.95a
17.67 ± 0.09b
58.67 ± 0.76b
StNPs/SA 74.45 ± 0.84a
95.06 ± 0.32a
101.74 ± 0.60b
15.75 ± 0.14d
55.24 ± 1.14c
StNPs/RS 72.70 ± 1.21ab
91.89 ± 0.66bc
103.49 ± 0.72a
16.46 ± 0.26c
64.09 ± 2.02a
Values represent the mean ± standard deviation of triplicate tests. Values in column having different superscripts (a, b, c, d) were significantly different (p 0.05).
StNPs represent starch nanoparticles, StNP/CS represent chitosan templeted or coating nanoparticles, GG represents Guar gum, GA represents Gum arabic, SA represents
Sodium alginate, RS represents Rosin.
X. Li et al. / Food Chemistry 229 (2017) 111–119 115

7. 3.4. X-ray diffraction pattern
All StNPs showed typical B-type XRD patterns with strong peaks
at 2h close to 5.6°, 15.3°, 17.1°, 22.5°, and 24.3° (Fig. S4), which was
in accordance with our previous reports (Sun et al., 2014). StNPs/
gum nanocomposites’ characteristic diffraction peaks intensity at
2h = 5.6°, 15.3°, and 17.1° increased significantly. The degree of
crystallinity of StNPs was 53.72%, higher than that of nanoparticles
using fractionated amylose (30.2–45.6%) and amylopectin (8.6–
9.2%) from potato starch reported by Qiu, Qin, Zhang, Xiong, and
Sun (2016), which indicated that StNPs fabricated from short-
chain glucan possessed a more compact structure. The degree of
crystallinity of the food gum-templated StNPs were also higher
than that of bare StNPs (Table 1). Interaction extent between
short-chain glucan, oriented arrange extent of the double helices,
numbers of double helices and crystal, and crystal size could cause
differences in crystallinity degree. The increasing crystallinity
degree of StNPs/gum nanocomposites could be caused by the
enhancement of interaction between short-chain glucan and the
oriented growth of crystal nuclei along the template. In addition,
the crystallinity degree of the StNPs/RS nanocomposites was
higher than that of the others, which indicated that most of the
RS served as a template in the growth of crystal nuclei, with few
coated on the surface of StNPs.
3.5. Fourier transform infrared spectra
To further understand why StNPs with different sizes and mor-
phologies could be successfully prepared using different food gums
as the template, FITR spectroscopy was applied to examine
whether there are interactions between SGCs and food gums. The
StNPs exhibited characteristic bands at 3305 cm1
(OAH stretch-
ing), 2918 cm1
(CAH stretching), 1647 cm1
(d (OAH) bending
of water), and 1353 cm1
(CH2) (Shi, Wang, Li, Benu, 2012).
The characteristic bands at around 1022, 1074, and 1156 cm1
were known as fingerprint region, which was caused by C-O ether
stretching vibration (Delval et al., 2004).
The FTIR spectra of five food gums were determined (data not
shown), though there are some differences among the FTIR spectra
of StNPs and five gums, StNPs and StNPs/food gum nanocompos-
ites had similar FTIR spectra. This could be due to the fact that
StNPs was supersaturated in this ratio of StNPs to food gums,
which is in good agreement with the recent reports (Assadpour,
Jafari, Maghsoudlou, 2017; Jafari, Sabahi, Rahaie, 2016).
Fig. S5 shows that the OH band of the StNPs was shifted to a lower
wavenumber when food gums were added, implying that hydroxyl
groups interaction between short-chain glucan and food gums
increased. The results suggested that hydrogen bonding between
StNPs and food gum molecules was a possible interaction in the
StNPs/gum nanocomposite system.
3.6. Loading capacity and encapsulation efficiency
Particle size, loading capacity, and encapsulation efficiency of
Sal B in StNPs/gum nanocomposites are listed in Table 2. Compared
with StNPs samples in the absence of Sal B, the particle size of
StNPs and StNPs/gum nanocomposites encapsulating Sal B showed
an increasing trend, which increased 59.6%, 32.7%, 72.1%, 17.7%,
80.5%, and 38.9%, respectively. However, their particle size was still
lower than 200 nm, which was in favor of increasing absorption
and the bioavailability of Sal B.
During the recrystallization of short-chain glucan, Sal B could be
tightly complexed inside short-chain glucan helices cavity. The
loading capacity and encapsulation efficiency of Sal B in StNPs
were 5.21% and 31.25%, respectively, which was higher compared
to values reported in the literature. Isacchi et al. (2011) developed
liposomes as Sal B carriers and determined encapsulation effi-
ciency of Sal B loaded conventional and PEGylated liposomes.
The results showed that conventional and PEGylated liposomes
had low encapsulation efficiency (24.84% and 22.72%). Peng et al.
(2010) reported that loading capacity of Sal B in phospholipid com-
plex loaded nanoparticles was only 3.21%. Our results showed that
most food gums improved StNPs’ loading capacity and encapsula-
tion efficiency (Table 2). In particular, the loading capacity of
StNPs/CS, and StNPs/GA nanocomposite could reach to 8.26% and
8.08%, respectively, because they can interact with StNPs through
hydrogen bonds and hydrophobic interactions to form a compact
covering layer, leading to embedding more Sal B. However, the
loading capacity of the StNPs/GG nanocomposites was weaker than
that of the others, which could be due to the strong adsorption of
GG onto the surface of nucleus and its structure with a high degree
of branching increasing the difficulty of the recrystallization to
form a relatively loose structure. The loose StNPs/GG nanocompos-
ites could not protect Sal B from removal after water washing.
What is more, the StNPs/GG nanocomposites’ size was large, which
made them easy to aggregate and led to difficulty regarding the
embedding of Sal B.
3.7. Morphology of Sal B loaded StNPs/gum nanocomposites
The morphology of the Sal B loaded StNPs/gum was also charac-
terized by TEM. As shown in Fig. 3, there are some changes in the
shape of the StNPs samples with the loading of Sal B. Compared
with the StNPs/gum nanocomposites, less or even no core-shell
structure could be observed, which was because the cavities of
the StNPs/CS and StNPs/GA nanocomposites were filled up by Sal
B; the StNPs/SA-Sal B nanocomposites became slenderer than the
spindly StNPs/SA nanocomposites, while spherical nanoparticles
including StNPs, StNPs/GG, and StNPs/RS nanocomposites had no
significant differences in shape before and after the embedding
of Sal B. Though the aggregation degree of Sal B loaded StNPs
nanocomposites decreased compared to that of StNPs, the shape
of the Sal B loaded StNPs nanocomposites became more irregular,
and their granules were not integrated and formed by the aggrega-
tion of many small nanoparticles. This could be because Sal B
inhibited the formation of hydrogen bonds between nanoparticles.
Xiao et al. (2013) also found that green tea polyphenols signifi-
cantly inhibited the retrogradation of rice, maize, and potato
starches.
3.8. In vitro release studies
Fig. 4 illustrates the Sal B release profiles from StNPs and StNPs/
gum nanocomposites in simulated gastrointestinal environment.
StNPs was found to strongly retain the loaded Sal B in gastric envi-
ronment. After 2 h incubation, only 21.42% Sal B was released. Such
slow release in the stomach is desirable for an oral carrier, because
Table 2
Encapsulation efficiency (EE) and loading capacity (LC) of starch nanoparticles (StNPs)
without or with different food gums for Salvianolic acid B (SaB) and their average
particle diameter (D).
Sample D (nm) EE (%) LC (%)
StNPs-SaB 154.7 ± 3.51b
(96.9)A
31.25 ± 0.58c
5.21 ± 0.07c
StNPs/CS-SaB 57.2 ± 1.68f
(43.1) 49.57 ± 2.09a
8.26 ± 0.25a
StNPs/GG-SaB 187.9 ± 3.19a
(109.2) 30.16 ± 0.44d
5.03 ± 0.24c
StNPs/AG-SaB 112.9 ± 4.23d
(95.9) 48.49 ± 1.30a
8.08 ± 0.16a
StNPs/SA-SaB 136.1 ± 0.96c
(75.4) 40.32 ± 1.51b
6.72 ± 0.28b
StNPs/RS-SaB 63.5 ± 1.03e
(45.7) 38.97 ± 1.34b
6.50 ± 0.19b
Values represent the mean ± standard deviation of triplicate tests. Values in column
having different superscripts (a, b, c, d) were significantly different (p 0.05). ()A
showed average particle diameters of StNPs and StNPs with different gums.
116 X. Li et al. / Food Chemistry 229 (2017) 111–119

8. there would be more Sal B available for absorption in the intestine.
Intestinal a-amylase plays an important role in controlling Sal B
release from StNPs because starch is apt to be hydrolyzed by a-
amylase in the small intestine. Sal B loaded StNPs displayed an ini-
tial burst release in simulated intestinal fluid, and then sustained
release in simulated colon fluid for over 18 h. After 24 h incubation
in different conditions, 94.05% Sal B was released from StNPs,
higher than the cumulative release amount (77%) of Sal B from gly-
cyrrhetinic acid compound liposomes (Lin et al., 2014). However,
the simulated digestive fluid in the report of Lin et al. (2014) was
only physiological saline.
Most StNPs/gum nanocomposites showed better capacities in
controlling the release of Sal B in simulated gastric fluid. This
was due to the fact that GA, GG, SA, and RS in media with pH values
lower than 6.5 had poor solubility and were hard to swell, leading
to their strong contraction on the surface of StNPs. In particular,
only 10.36% Sal B was released from StNPs/SA nanocomposites
after incubated in simulated gastric fluid. Similar findings were
previously reported by Huang et al. (2014), who fabricated core-
shell gelatin-alginate composite microparticles, and found that
they could remain intact in gastric juice for at least 3 h, indicating
that the gelatin core could be well protected by alginate shell in
Fig. 3. TEM images of starch nanoparticles without (A) or with chitosan (B), Guar gum (C), gum arabic (D), sodium alginate (E) and Rosin (F) loaded with salvianolic acid B.
0 4 8 12 16 20 24
0
20
40
60
80
100
pH=7.4
pH=6.8
Cumulative
Release
(%)
Time (h)
StNPs
StNPs/CS
StNPs/GG
StNPs/GA
StNPs/SA
StNPs/RS
pH=1.2
Fig. 4. Cumulative release profile of salvianolic acid B loaded starch nanoparticles
(StNPs) and StNPs/gum nanocomposite in simulated gastric fluid (pH = 1.2),
intestinal fluid (pH = 6.8), and colon fluid (pH = 7.4).
X. Li et al. / Food Chemistry 229 (2017) 111–119 117

9. acid environment. Sal B release was greater from StNPs/CS than
StNPs in simulated gastric fluid (Fig. 4), which could be attributed
to the swelling of CS in acidic condition and breakdown of hydro-
gen bonds between StNPs and CS. However, the release of Sal B
showed a relatively slower releasing rate in most of StNPs/gum
nanocomposites than that in StNPs, which indicated the coating
of gums (except for CS) could significantly improve the capacity
of StNPs in controlling release of Sal B.
4. Conclusions
we have successfully complexed food gums (as a template) with
short-chain glucan to form StNPs/food gum nanocomposites with
controlled size to encapsulate salvianolic acid B (Sal B) and found
that biopolymer nanoparticles could be stable against pH, salt,
and high temperature. In addition, they showed different
morphologies such as spherical, spindly, and core-shell shapes.
Particularly StNPs/CS nanocomposites and StNPs/GA nanocompos-
ites, with hollow or solid nanoparticle structure, could be used as a
promising delivery system for nutrients or drugs in physiological
conditions. Additionally, our results demonstrate that StNPs/gum
nanocomposites performed well in carrying and controlled releas-
ing Sal B in artificial intestinal juice and blood fluid and are suitable
for delivering active compounds to blood fluid.
Acknowledgment
The study was supported by the National Natural Science Foun-
dation, China (Grant No. 31671814).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.foodchem.2017.
02.059.
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