Biochemical and Biophysical Research Communications 516 (2019) 1175e1182

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Biochemical and Biophysical Research Communications

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

Structural characterization and emulsifying properties of thinned-

young apples polysaccharides
Yanjie Hou, Tian Gong, Jiangtao Zhang, Xi Yang*, Yurong Guo**
College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi'an, 710062, PR China

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

Article history: The thinned-young apple polysaccharides from three varieties were obtained by hot water extraction at
Received 1 July 2019 88 C for 120 min. The compositional monosaccharides of the three polysaccharides were shown to be the
Accepted 5 July 2019 same (xylose, mannose, galactose and glucose) and the molecular weights of the polysaccharides were in
Available online 8 July 2019
the range of 200e300 kDa. Compared with “Qinyang” and “Pinklady”, the polysaccharide from “Jinshiji”
had the highest emulsifying capacity. Moreover, the variations in pH and cation ion concentrations had
Keywords:
also a significant effect on the emulsifying properties of the extracted polysaccharides. At pH 2.0e4.0, the
Thinned-young apple
prepared emulsion had smaller droplet sizes than at higher pH values. Although the emulsion was stable
Polysaccharide
Emulsifying properties
at low concentrations of Naþ and Ca2þ ions, high concentrations of Naþ and Ca2þ led to significant
Rheological properties destabilization of the emulsion. Conclusively, our results demonstrated the potential application of
thinned-young apple polysaccharide as a natural polysaccharide emulsifying agent.
© 2019 Elsevier Inc. All rights reserved.

1. Introduction as a novel a-amylase inhibitor [5]. On the other hand, thinned-

Fruit thinning is a key step to ensure the size, color and quality of
apple after harvest [1]. Annually, fruit thinning produces approxi-
mately 1.9 million tons of young apples which are unutilized in
China [2]. Most of the thinned-young apples are directly discarded
in the grove and become a good source for microbial reproduction,
including Alternariamali, Physalosporapiricola and Glomer-
ellacigulata, etc. As a consequence, it increases̊ the risk of microbial
infections and may lead to a significant reduction in fruit quality
and yield. In recent years, thinned-young apples have been
increasingly reported to contain numerous phytochemicals and
biological active compounds. For instance, it has been verified that
thinned-young apple polyphenols possessed significant antioxi-
dant, anticancer, antiallergenic and antibacterial properties [3,4].
Specially, thinned-young apple polyphenols had good inhibitory
effects on a-amylase, which pinpointed their potential application
* Corresponding author. College of Food Engineering and Nutritional Science,
Shaanxi Normal University, 620 West Chang'an Avenue, Chang'an, Xi'an, 710119, PR
China.
** Corresponding author. College of Food Engineering and Nutritional Science,
Shaanxi Normal University, 620 West Chang'an Avenue, Chang'an, Xi'an, 710119, PR
China.
E-mail addresses: yangxi@snnu.edu.cn (X. Yang), yrguo730@snnu.edu.cn
(Y. Guo).
young apples also contained ~10% of polysaccharides (w/w, dry
basis) according to our previous works. And further functional as-
says showed that thinned young apple polysaccharide exhibited
good antioxidant activities and exerted significant alleviating ef-
fects on hepatic metabolic disorder in high fat diet (HFD)-induced
obese mice [2,3,6]. Hence, it is promising to extract natural phy-
tochemicals from thinned-young apples.
An emulsion is a dispersion of one liquid dispersed into another
immiscible liquid. In the food industry, there are numerous emul-
sifiers available when preparing an emulsion, including sucrose
ester, fatty glyceride, some proteins and polysaccharides, etc. Based
on the sources, emulsifying agents can be grouped into two cate-
gories: natural emulsifiers and artificial emulsifiers. In the last
decades, natural emulsifiers have attracted increasing attention on
account of their nontoxicity, low cost and accessibility from
renewable resources, etc. As an example, natural polysaccharides,
especially gum arabic and sugar beet pectin, have been successfully
applied in the beverage industry [7e10]. In general, a poly-
saccharide that can be used as emulsifying agent must necessitate
the following criteria: (i) they can adsorb at the oil-water interface;
(ii) they can protect the emulsion droplets against immediate
recoalescence. In theory, all polysaccharides that possess the
interface-adsorbing and emulsion stabilizing properties can be
used as natural emulsifying agents.
In this study, three thinned-young apple polysaccharides

https://doi.org/10.1016/j.bbrc.2019.07.019
0006-291X/© 2019 Elsevier Inc. All rights reserved.
1176 Y. Hou et al. / Biochemical and Biophysical Research Communications 516 (2019) 1175e1182
(“Pinklady”, “Qinyang” and “Jinshiji”) were extracted by hot water,
and then the polysaccharide characteristics were characterized in
terms of their basic compositions, molecular weight and mono-
saccharide compositions. Besides, their structural properties were
further determined by FTIR, NMR spectroscopy, XRD pattern and
thermal analysis (TG/DTG). To explore the emulsifying properties of
thinned-young apple polysaccharides, the emulsions based on 2.0%
polysaccharides solution (w/v, %) were prepared and the effects of
pH, cation type (Naþ, Ca2þ) and concentrations on the emulsions
were also investigated, in order to provide references for searching
natural polysaccharide emulsifier.
2. Materials and methods
2.1. Extraction of thinned-young apple polysaccharides
Thinned-young apples (“Pinklady”, “Qinyang” and “Jinshiji”)
were collected 40 days after blossom from Baishui test station
(Shaanxi province, China). All the fruits were sliced (1.0 mm in
thickness) and immediately immersed into a hot water bath at 95 C
for 15 s. Then the slices were dried at 55 C for 36 h and ground into
powder form, and sieved by a 60 mesh sieve. The polysaccharide
extraction was carried out according to the method of Dou, Meng,
Liu, Li, Ren and Guo [11] with some modifications. The powder was
mixed with 10-fold volumes of 95% ethanol (w/v) and the mixture
was refluxed at 80 C for 6 h to remove phenolic and lipophilic
substances. After filtrating, the residues were dried at 55 C until
constant weight and then mixed with 41-fold volumes of deionized
water for polysaccharide extraction (88 C for 120 min). Subse-
quently, the supernatant was obtained by centrifuging at 4500g
for 20 min and concentrated to approximately a quarter of the
original volume of the solution. The concentrated solution was
mixed with four-volume of 95% ethanol (v/v, %) for polysaccharide
precipitation (4 C, 24 h). After centrifuging the mixture solution at
4500g for 20 min, the precipitate was obtained and then dis-
solved into deionized water and subjected to rotary evaporation at
60 C for 1 h to fully remove the residual ethanol. To remove the
oxidated polyphenols that may have adverse effects on the
following emulsion observation, the polysaccharide solution was
eluted using X-5 macroporous resin. During the run, the poly-
saccharide concentration was 3.0 mg/mL and the flow rate was
2 mL/min. Afterwards, the polysaccharide was obtained by
lyophilization.
̊ from 13。to 65。 with a step of 0.02。by a Powder X-ray Diffrac-
̊ tometer (D8 Advance, Germany).
2.2. Composition identification of thinned-young apple
polysaccharides
The total polysaccharide content was determined by the
phenol-sulfuric acid̊ colorimetric method [12]. Different concen-
trations of glucose solutions (0e90 mg/mL) were used as standards
̊
to establish a standard curve. The content of phenolic compounds
in polysaccharides was determined according to the Folin-
̊
Ciocalteu method [13]. Briefly, 2 mL of 0.2 mg/mL polysaccharide
solution was mixed with 2.0 mL Folin-Ciocalteu reagent (2 N,
Sigma) and 24.0 mL 7.5% of sodium carbonate solution and then
the solution volume was diluted to 50 mL with deionized water.
Subsequently the̊ mixture was incubated at room temperature for
2 h and the absorbance of the mixture was determined at 765 nm.
Furthermore, 0.2 mg/mL thinned-young apple polysaccharide was
scanned
̊ within the range of 190e400 nm by a Hitachi U-3010 UV
spectrophotometer (HITICHI Company, Japan). Meanwhile,
deionized water instead of polysaccharide solution was taken as a
control.
2.3. Characterization of thinned-young apple polysaccharides
2.3.1. Molecular weight
The method of Liang [14] was used with some modifications to
determine the molecular weight of the polysaccharides. 1.0 mg/mL
polysaccharide solution was prepared and filtered through a
0.45 mm filter membrane. Then, the solution was injected into a
high performance size exclusion chromatography (HPSEC), during
which 0.05 M NaNO3 was used as mobile phase and the flow rate
was 0.7 mL/min. The molecular weight was estimated according to
the standard curve, which was established by the Dextrans stan-
dard of T-10, T-40, T-70, T-150, T-500, T-1000 and T-2000.
2.3.2. Determination of monosaccharides composition
4.0 mL of 3 M TFA was mixed with 80 mg of polysaccharide, and
the solution was hydrolyzed on an oil bath at 110 C for 4 h to release
constitutive monosaccharidesThe hydrolyzed solution was cooled
to room temperature, after which 3.0 mL of methanol was added
into the hydrolyzed solution to remove the residual TFA. Then, the
hydrolysate was derived and injected into a gas chromatography-
mass spectrometer (GC-MS) for composition analysis [15]. During
the procedure, the column temperature was kept at 120 C for 1 min
and was then linearly increased to 240 C within 49 min at a heating
rate of 2.5 C/min, after which the column temperature was kept at
240 C for 7 min. The pure helium with a flow rate of 1.0 mL/min was
served as the carrier gas.
2.3.3. FTIR and NMR analysis
About 10 mg of thinned-young apple polysaccharide was mixed
with dried KBr (1:100) and pressed into a semitransparent pellet
for FTIR analysis. The spectrum within the range of 4000 to
400 cm1 was recorded by using an IR spectrometer (Tensor27,
Bruker, Germany).
The freezing-dried polysaccharides were dissolved in D2O to
achieve a concentration of 40 mg/g. Then, the solutions were
determined by using a Bruker 600 MHz NMR spectrometer (Bruker,
Rheinstetten, Germany), during which the 1H and 13C NMR spectra
were recorded.
2.3.4. X-ray diffraction ̊
X-ray diffraction was used to measure the crystalline structures
of thinned-young apple polysaccharides. Before the test, the poly-
saccharides was dried at a oven at 45 C overnight to remove the
residual moisture, after which the polysaccharide was scanned
̊
̊
̊
2.3.5. Thermal analysis (TG/DTG)
̊Thermal analysis was performed to investigate the thermal
properties of thinned-young apple polysaccharides. About 10 mg
polysaccharide was heated from 25 to 600 C with a heating rate of
10 C/min by using a thermalgravimetric analyzer (Q6000SDT, TA
Instrument, USA). Meanwhile, pure nitrogen served as protective
gas and an empty aluminum pan as the reference.
2.4. Emulsion preparation
Emulsion preparation was carried out based on the method
reported by Schmidt, Schütz and Schuchmann [16] with some
modifications. 5.0 mL of the polysaccharide solution (2.0%, w/v)
was mixed with 1.25 mL colleseed oil by a rotor homogenizer (T25
digital ultra-turrax, IKA, Germany) at 20000 rmp for 120 s for
emulsion preparation. In the meantime, the deionized water
instead of polysaccharides solution was taken as the control.
̊
 Y. Hou et al. / Biochemical and Biophysical Research Communications 516 (2019) 1175e1182
2.5. The influence of pH, cation type and concentrations on
emulsions
To investigate the emulsifying properties of thinned-young ap-
ple polysaccharides, the emulsions at various pH values, cation
concentrations (Naþ and Ca2þ) were prepared according to the
method mentioned above. The pH values of polysaccharide solu-
tions were adjusted by 0.5 M NaOH and 0.2 M HCl solutions to 2.0,
4.0, 6.0 and 8.0, respectively. Similarly, different concentrations of
Naþ and Ca2þ were added into 2.0% polysaccharides solution to
achieve gradient concentrations of ionic environment (20, 40, 60,
80 mg/g, respectively). Then, the emulsions were prepared by ho-
mogenizing the mixture of 5.0 mL of polysaccharide solution and
1.25 mL of colleseed oil, as described earlier.
2.6. Rheological properties of the emulsions
The rheological properties of the emulsions were investigated
by a stress-controlled rheometer (AR-G2, TA Instruments, USA)
equipped with a parallel plate (40 mm, 1 mm gap). After ~2.5 mL of
emulsion was loaded onto the sample stage, the viscosity of the
emulsion within the shear rate of 0.1e600 s1 was recorded.
2.7. Microscopic observation of the emulsions
Before the observation, the staining solutions were prepared
according to Yang, Nisar, Hou, Gou, Sun and Guo [17]. Briefly, Nile
red was dissolved into 1,2-propylenge glycol to achieve a concen-
tration of 0.1% (w/v) and the staining solution was held at 4 C for
24 h for equilibrium. Then, the emulsion containing 30 mL staining
solution was prepared as described earlier. After that, ~40 mL of
emulsion was smeared on a glass slide, followed by covering with a
cover slip, and immediately the emulsions were observed and
imaged by a confocal laser scanning microscope (CLSM, FV1200,
Olympus Corporation, Japan) in the fluorescence model and at
595 nm of excitation wavelength.
2.8. Statistical analysis
All the data presented in the study were analyzed by SPSS 18.0
software and the results were expressed as the mean ± standard
deviation (SD). All the figures were plotted using Origin 8.0
software.
3. Results and discussion
3.1. Compositions of thinned-young apple polysaccharides
We firstly analyzed the compositions of the extracted poly-
saccharides, and found that the total polysaccharide contents of
“Pinklady”, “Qinyang” and “Jinshiji” were 92.6, 91.54, and 93.57%,
respectively. Moreover, no polyphenols were detected in the
polysaccharides. As shown in Fig. 1A, the UV spectra showed ̊ that
there was no significant absorption peak occurring at around
260e280 nm, indicating no protein lingering in the
polysaccharides.
3.2. Molecular weight and monosaccharide compositions
The molecular weights of three thinned-young apple poly-
saccharides are displayed in Fig. 1B, and the detailed molecular
parameters are summarized in Table 1. As shown in Fig. 1B, three
polysaccharides exhibited similar molecular distribution curves,
but “Jinshiji” polysaccharide had the highest molecular weight
(293 kDa), followed by “Pinklady” (270 kDa) and “Qinyang”
1177
polysaccharide (207 kDa). Besides, the ratio of Mw to Mn can also be
calculated to indicate the polydispersity index of polysaccharides. A
higher polydispersity index represents that the polysaccharide
molecules exist more dispersed in aqueous solutions [18]. From
Table 1, we found that three polysaccharides possessed similar
polydispersity indexes.
Moreover, GC-MS was applied to determine the mono-
saccharide compositions of thinned-young apple polysaccharide,
and the results are listed in Table 2. It was found that three poly-
saccharides had the same monosaccharide compositions: xylose,
mannose, galactose and glucose. Of those monosaccharides,
glucose and mannose were the major compositions, which
accounted for ~93% of total monosaccharides in “Qinyang”, fol-
lowed by ~87.9% for “Jinshiji” ~83% for “Pinklady”.
3.3. FTIR analysis
As shown in Fig. 2A, the broad absorption peaks appearing at
3377 cm1 for “Pinklady”, 3381 cm1 for “Qinyang” and 3392 cm1
for “Jinshiji” were attributed to the eOH stretching vibrations, and
the weak peaks occurring at 2931 cm1 were caused by the
stretching vibration of CeH of the eCH2 groups. Besides, the ab-
sorption peaks at approximately 1740 cm1 were attributed to the
C¼O stretching vibration of ester carbonyl. Compared with “Jin-
shiji” and “Pinklady”, “Qinyang” did not show a discernible ab-
sorption peak at near 1741 cm1, indicating that “Qinyang”
polysaccharide was almost no esterified. The peaks at around
1610 cm1 and 1410 cm1 were assigned to C¼O stretching vibra-
tion of non-methylated carboxyl and COO- asymmetric stretching
vibration, respectively [19]. Moreover, the absorption band at
around 1250 cm1 and 1076 cm1 were corresponded to the vi-
bration of CeO glycosidic bond. Specially, the broad absorption
peak at around 600 cm1 indicated that polysaccharides may
contain skeletal models of pyranose unit. In addition, the absorp-
tion peak at 885 cm1 indicated the presence of b-type glycosidic
bond [20,21]. Generally, the esterified carboxyl groups in poly-
saccharides are able to stabilize emulsions by adsorbing onto the
oil-and-water interface [22]. Therefore, the amount of hydrophobic
groups is an important parameter when assessing the emulsifying
capacity of polysaccharides [23]. In our study, both of “Pinklady”
and “Jinshiji” had absorption peaks at 1741 cm1, indicating that the
polysaccharides contained esterified carboxyl groups, which may
be favorable for their emulsifying properties.
1 13
3.4. H and C NMR spectra
1
H and 13C NMR spectra were used to explore polysaccharide
structures. The spectra of three polysaccharides are essentially
similar, and therefore, “Jinshiji” was selected as a representative to
present NMR spectra, as shown in Fig. 2B&C. The elucidation of 1H
and 13C NMR spectra of “Pinklady” and “Qinyang” were given as
supplemental materials. It was observed from Fig. 2B that the sig-
nals from 3.3 to 5.1 ppm were the characteristic signals of poly-
saccharides [24]. The signals appearing at 5.50e4.90 ppm and
4.90e4.30 ppm suggested the polysaccharides had a- and b-
glycosidic linkages, respectively [25]. These signals corresponded to
the anomeric carbon signals at the region of 10290 ppm in 13C
NMR. The result was consistent with the FTIR analysis. Besides, the
signal of 4.50-4.00 was assigned to the ring proton region. The
signals of 3.94, 3.84 and 3.16 were attributed to the presence of the
b-D-xylose. The signals at around 5.1 and 4.5 ppm were attributed
to a-D-glucose and b-D-mannose, respectively [24]. In addition,
four signals were found to locate at the region of 102-92 ppm
(Fig. 2C), indicating the presence of b-D-xylose, b-D-glucose, a-D-
mannose and a-D-galactose [26]. The signal at 80.7 ppm suggested
1178 Y. Hou et al. / Biochemical and Biophysical Research Communications 516 (2019) 1175e1182

Fig. 1. UV spectra of thinned-young apple polysaccharides (A) and molecular weight distribution curves of thinned-young apple polysaccharides (B).

Table 1 attributed to the evaporation of the moisture absorbed in poly-

Molecular weight distribution parameters of thinned-young apple polysaccharides. saccharides. The second weight loss occurred at around 150 C,
Samples Peaks 1 2 3 which may be due to the loss of the bounded water [30]. The third
mass loss occurred at the range of 200e350 C, which corresponded
“Pinklady” Mn (kDa) 44.76 0.76 0.13
Mw (kDa) 270.86 0.89 0.19 to the decomposition of polysaccharides. During the third step,
Mw/Mn 6.05 1.17 1.46 “Pinklady”, “Qinyang” and “Jinshiji” polysaccharides lost about 47,
“Qinyang” Mn (kDa) 33.87 0.71 0.13 46 and 48% of their original weights, respectively. The results
Mw (kDa) 207.75 0.82 0.19
suggested that “Qinyang” had a higher thermal stability than
Mw/Mn 6.13 1.15 1.46
“Jinshiji” Mn (kDa) 48.55 0.77 0.13
“Pinklady” and “Jinshiji”.
Mw (kDa) 293.19 0.90 0.20
Mw/Mn 6.04 1.17 1.54
3.6. Emulsifying properties and microscopic observation of the
emulsions

Table 2
Monosaccharide constituents of thinned-young apple polysaccharides.
Samples “Pinklady” “Qinyang”
Xylose 6.9 3.8
Mannose 36.4 32.7
Galactose 9.1 3.5
Glucose 47.4 60.0
that the position C-3 connecting by 1 / 3 linked signal of glucose
residues and 1 / 6 linked monosaccharides backbone existed in
thinned-young apple polysaccharides [27]. Unlike “Qinyang”
polysaccharide, “Pinklady” and “Jinshiji” had a significant signal at
174 ppm, which was due to the presence of eCOOH groups [28].
3.5. XRD and TG/DTG
The crystalline structure of thinned-young apple poly-
saccharides can be caused by the partial alignment of poly-
saccharides molecules [29]. As displayed in Fig. 3A, the results
showed there were semi-crystalline and amorphous structures
existing in thinned young apple polysaccharides [30]. Both “Jin-
shiji” and “Pinklady” exhibited two broad peaks at the 2q of
20.2。and 30.5。, respectively, whereas “Qinyang” only had one
peak occurred at 20.2。. These results indicated that “Qinyang” may
have a lower crystallinity.
Thermal analysis (TG/DTG) is performed to investigate the
thermal properties of thinned-young apple polysaccharides. As
shown in Fig. 3B&C, the polysaccharides exhibited three main
weight loss steps. The first step occurred below 100 C, which was
After preparing emulsions, we firstly investigated the rheolog-
ical properties of the emulsions at different ionic environments, pH
“Jinshiji” values etc. As shown in Fig. 4A&B, all the emulsions exhibited the
5.99 characteristics of non-Newtonian fluids, with significant shear-
26.2 thinning properties [31]. The viscosity decreasing trends for all
6.47
emulsions were the same and conformed to the order of “Jin-
61.7
shiji” > “Pinklady” > “Qinyang”, and therefore, “Jinshiji” was
selected as a represent to explore the effects of cation ions (Naþ and
Ca2þ). The viscosity of “Pinklady” and “Qinyang” were provided in
the supplementary materials. The incorporation of low amounts of
Naþ (20 and 40 mg/g) increased the viscosity of the emulsions,
which was attributed more compact molecular conformation of the
̊
polysaccharide induced by Naþ. Comparing with Naþ, Ca2þ signif-
icantly increased the viscosity of the emulsions, which may be
̊
related with the formation of gel-like network [31]. Generally, Naþ
had a greater shielding effect on the polysaccharide chains by
collapsing down the side chains [32]. However, Ca2þ could facilitate
the formation of cross links by combining with the dissociative
carboxylic acid groups in polysaccharide chains [31,33], thereby
increasing the emulsions viscosity. It is worth noting that the
emulsion viscosity was decreased with the increasing cation
addition (Naþ and Ca2þ). It is understood by the fact that high
strength of ionic environments diminished the electrostatic re-
pulsions between polysaccharides molecules, and even caused
desorption of polysaccharide molecules from the droplet surface
[34].
Moreover, the viscosity of the prepared emulsions at various pH
values ranging from 2.0 to 8.0 are displayed in Fig. 4C. Generally, pH
values can influence the emulsion viscosity by altering the molec-
ular conformation of polysaccharides. At pH ¼ 2.0 and 4.0, the
Y. Hou et al. / Biochemical and Biophysical Research Communications 516 (2019) 1175e1182 1179

Fig. 2. FTIR (A), 1H (B) and 13

C (C) NMR spectra of thinned-young apple polysaccharides.
1180 Y. Hou et al. / Biochemical and Biophysical Research Communications 516 (2019) 1175e1182

Fig. 3. X-ray (A), TG (B) and DTG (C) curves of thinned-young apple polysaccharides.

polysaccharide molecules that adsorbed onto the surfaces of the

emulsion droplets were less dissociated, and therefore had a
compact conformation. That is favorable for preventing emulsion
droplets from coalescence [35]. At pH ¼ 6.0 and 8.0, the emulsion
viscosity was decreased, which may be because polysaccharide
molecules had a looser conformation at high pH values. On the
other hand, at high pH values, the carboxyl groups in the poly-
Fig. 4. The viscosity of polysaccharide solution and the resulting emulsions at different
conditions. (A): different Naþ ion concentrations (20, 40, 60 and 80 mg/g poly-
saccharides); (B): different Ca2þ ion concentrations (20, 40, 60 and 80 mg/g poly-
saccharides); (C): different pH values (2.0, 4.0 6.0 and 8.0).

saccharides were more dissociated, causing increased intra-

molecular repulsions, weakening the strength of hydrated layer on
the droplet surface. Thus, at high pH value, the emulsions were
unstable, and small emulsion droplets tended to merge to form a
larger droplet [17,36].
The oil droplets of the emulsions were imaged by CLSM, and the
results are presented in Fig. 5. It was found that “Jinshiji” had the
smallest droplet size, as reflected in Fig. 5A, demonstrating its good
emulsifying capacity. At pH ¼ 2.0 and 4.0, the emulsion droplets
were uniform, whereas the droplets coalescence occurred when
 Y. Hou et al. / Biochemical and Biophysical Research Communications 516 (2019) 1175e1182 1181

the pH increased to above 6.0 (Fig. 5B). Especially, when the pH

increased to 8.0, the coalescence of droplets became very signifi-
cant. Moreover, at low concentrations of Naþ (below 20 mg/g) and
Ca2þ (below 40 mg/g), the emulsion droplets were uniform and had
significant smaller sizes (Fig. 5C and D). However, as the ionic
concentration increased, the droplets coalescence was observed
and the droplet sizes were beyond 100 mm.
In summary, three thinned young apple polysaccharides were
obtained by hot water extraction, and composition monosaccharide
analyses suggested that they had the same monosaccharide con-
stituents but varying molar ratios. In detail, all three poly-
saccharides were composed of xylose, mannose, galactose and
glucose, among which glucose and mannose accounted for ~93% of
total monosaccharides for “Qinyang”, ~87.9% for “Jinshiji” and ~83%
for “Pinklady”, respectively. The polysaccharide from “Jinshiji” had
better emulsifying capacity than other two polysaccharides. At pH
2.0e4.0, the emulsions had uniform and smaller droplets than at
high pH values. Besides, at low concentrations of Naþ, Ca2þ, the
emulsion exhibited good stability, whereas high concentrations of
Naþ, Ca2þ led to significant de-emulsification of the emulsion.
Conclusively, our results may provide information for searching
natural polysaccharide emulsifying agents to be used in the food
industry.

Conflicts of interest

We declare that we have no personal relationships with other

people and organizations that influence our work. There is no
commercial or associative interest that represents a conflict of in-
terest in connection with the work submitted.

Acknowledgement

This work is financially supported by China Agricultural

Research System (CARS-28) from 584 Ministry of Agriculture of the
People's Republic of China.

Appendix A. Supplementary data

Supplementary data to this article can be found online at

https://doi.org/10.1016/j.bbrc.2019.07.019.

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