International Journal of Biological Macromolecules

Natural polysaccharide from buckwheat flower and its emulsifying property under high
oil phase condition
--Manuscript Draft--

Manuscript Number: IJBIOMAC-D-23-11611

Article Type: Research Paper

Section/Category: Carbohydrates, Natural Polyacids and Lignins

Keywords: Natural polysaccharide from buckwheat flower and its emulsifying property under high
oil phase condition

Abstract: A basis for the future utilization of natural buckwheat flower polysaccharide (BFP) from
discarded buckwheat flowers was provided by this experiment, which studied the
characterization and emulsifying properties of BFP as emulsifier under high oil phase
conditions. The physico-chemical properties of BFP were determined and was found
that BFP was hydrophilic with an average molecular weight of 2.421×105 Da, and also
with β-D-pyranose glucose ring. The interfacial tension of BFP decreased rapidly with
the start of the experiment, and decreased with the increase of BFP
concentration, showing the potential of BFP as an emulsifier. By exploring the effects
of different oil phases and different concentrations of BFP on emulsion stability, it was
shown that the emulsion stability was strongest at the concentration of 1.0wt% BFP
and 60.0wt% of oil phase. Under such concentration, BFP’s high oil phase emulsion
showed good heat resistance, acid resistance, and ion resistance on emulsion stability.
Furthermore, in the simulated digestion experiment delivering curcumin, the
bioavailability of curcumin in BFP’s high oil phase emulsion was 27.05%, significantly
higher than 7.10% for direct digestion of curcumin, proving a high application value in
the field of high oil phase emulsions by BFP.

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Declaration of Interest Statement

Declaration of interest statement

The authors declare that we have no known competing financial interests or
personal relationships that could have appeared to influence the work reported in this
paper.
Abstract

Abstract

A basis for the future utilization of natural buckwheat flower polysaccharide

(BFP) from discarded buckwheat flowers was provided by this experiment, which
studied the characterization and emulsifying properties of BFP as emulsifier under
high oil phase conditions. The physico-chemical properties of BFP were determined
and was found that BFP was hydrophilic with an average molecular weight of
2.421×105 Da, and also with β-D-pyranose glucose ring. The interfacial tension of
BFP decreased rapidly with the start of the experiment, and decreased with the
increase of BFP concentration, showing the potential of BFP as an emulsifier. By
exploring the effects of different oil phases and different concentrations of BFP on
emulsion stability, it was shown that the emulsion stability was strongest at the
concentration of 1.0wt% BFP and 60.0wt% of oil phase. Under such concentration,
BFP’s high oil phase emulsion showed good heat resistance, acid resistance, and ion
resistance on emulsion stability. Furthermore, in the simulated digestion experiment
delivering curcumin, the bioavailability of curcumin in BFP’s high oil phase emulsion
was 27.05%, significantly higher than 7.10% for direct digestion of curcumin, proving
a high application value in the field of high oil phase emulsions by BFP.
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1 1 Natural polysaccharide from buckwheat flower and

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4 2 its emulsifying property under high oil phase
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9 4 Yu Zhanga*, Yanghui Xua, Tianqi Fanga, Qin Qiua, Maobin Chena
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Key Laboratory of Fermentation Engineering, Ministry of Education; National “111” Center for
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20 9 Cellular Regulation and Molecular Pharmaceutics; Hubei Key Laboratory of Industrial
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22 10 Microbiology; School of Biological Engineering and Food, Hubei University of Technology, Wuhan
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24 11 430068, China
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14 Keywords: buckwheat flower; polysaccharide; high oil phase emulsion; simulated
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15 digestion
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 1 17 1.Introduction
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5 18 Buckwheat is a crop that is widely grown around the world and is one of our
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7 19 important sources of food (Zhao et al., 2018). It contains high biological value, balanced
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9 20 amino acid composition of protein, fiber, vitamin B1 and B2, zinc, copper, manganese
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11 21 and selenium (Lee et al., 2010). Studies have shown that buckwheat flowers are rich in
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13 22 polysaccharides, similar to other natural plant polysaccharides (Zhu et al. 2018), and
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15 23 have good application prospects in the fields of food, medicine, cosmetics, etc. In
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17 24 addition to various nutrients, buckwheat polysaccharides have a variety of functional
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19 25 activities, such as hypolipidemic, antioxidant, antitumor, neuroprotective activities and
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21 26 immunoregulatory, with no toxic side effects (Ji et al., 2019). At present, flowers as
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23 27 agricultural residues are commonly used for agricultural purposes such as organic
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25 28 fertilizers, feed purposes such as feed additives, food production and so on. Compared
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27 29 with the synthetic emulsifiers used in the market, such as sorbitol, fatty alcohol ethoxide
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29 30 and sucrose ester, which can cause certain pressure on intestinal absorption, studies
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31 have shown that buckwheat polysaccharide can alleviate intestinal absorption pressure
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32 in mice (Zhu et al. 2018). Although there are already some measures for the application
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36 33 of flower waste, there is little research on the utilization of the polysaccharides in
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38 34 buckwheat flowers (Ne š ovi ć et al., 2021). Considering the biological activity of
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40 35 buckwheat flower polysaccharides and the possibility of being used as a food emulsifier,
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42 36 this is a great waste of resources. Polysaccharide can be extracted from buckwheat
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44 37 flowers by using hot water as extractant, which is simple, low cost with high extraction
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46 38 rate, and is suitable for reuse of waste buckwheat flowers. So extracting
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48 39 polysaccharides from buckwheat flowers for exploration can reduce the waste of social
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50 40 resources and also generate higher economic benefits.
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52 41 Oil-in-water emulsions are thermodynamically unstable systems. As the storage
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54 42 time increases or the external conditions change, the emulsion system will experience
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56 43 flocculation, aggregation, and stratification, causing the entire emulsion stability
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58 44 system to become unbalanced (Tao et al., 2019). Therefore, an emulsifier with good
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 45 emulsifying properties and stability is very important (Chen et al., 2019). In the food
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46 processing process, common emulsifiers are divided into three categories:
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47 macromolecular proteins, polysaccharides, and small molecule surfactants. Emulsifiers
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7 48 have different properties, and their emulsifying ability and the properties of the
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9 49 prepared emulsions also vary greatly (Gao et al., 2021). Compared with conventional
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11 50 emulsions and nanoemulsions, high oil phase emulsions (oil phase ratio ≥50.0wt%)
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51 often have larger particle sizes and can carry more when embedding oil-soluble
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16 52 nutrients without weakening the protective effect on nutrients (Calderón-chiu et al.,
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18 53 2021; Schädle et al., 2022).
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20 54 In this experiment, buckwheat flower polysaccharides (BFP) were prepared from
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22 55 waste buckwheat flowers by water extraction. Firstly, the physical properties of BFP
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24 56 were characterized, and then a high oil phase emulsion was prepared for its better
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26 57 emulsifying properties. By studying the stability of BFP high oil phase emulsion under
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28 58 different conditions, it was found that BFP high oil phase emulsion has good resistance.
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30 59 The protective effect of BFP emulsion on curcumin in simulated gastrointestinal fluid
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32 60 was also explored, and was found to be obvious. The results of this experiment can be
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34 61 used for the future development of BFP in the fields of food, medicine, cosmetics, etc.,
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36 62 and also provide support for further research on BFP.
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63 2.Materials and Methods
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64 2.1. Materials
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47 65 Buckwheat flowers were purchased from Xi'an Guohao Biological Company,
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49 66 Medium chain triglyceride (MCT) was purchased from commercial food suppliers
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67 (Mazola, ACH Food Companies, Memphis, TN, USA), mannose (Man), ribose (Rib),
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54 68 rhamnose (Rha), glucuronic acid (GlcA), galacturonic acid (GalA), N-acetyl-
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56 69 glucosamine, glucose (Glc), n-acetyl-galactose, galactose (Gal), xylose (Xyl),
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58 70 arabinose (Ara), fucose (Fuc), nile red, sodium hydroxide, and other standard products
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60 71 such as hydrochloric acid are all purchased from Sigma-Aldrich (St. Louis, MO, USA)
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 1 72 2.2. Extraction of BFP
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4 73 Buckwheat flowers were ground and extracted by continuous stirring for 5 hours
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6 74 with hot water (1:30m/v) at 90°C. After centrifuging at 8000rpm for 5 minutes, the
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8 75 supernatant was concentrated under reduced pressure at 50°C using a rotary evaporator.
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10 76 Three times the volume of ethanol was added to the concentrated solution, and the
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12 77 polysaccharide was precipitated overnight at 4°C. The polysaccharide was redissolved
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14 78 in water and concentrated again according to the above steps to remove residual ethanol,
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16 79 and then vacuum freeze-dried. The crude polysaccharide prepared by the above steps
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18 80 was dissolved in double distilled water, and 1/4 volume of Sevag reagent was added to
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20 81 remove protein, stirred at 160 rpm for 25 min, centrifuged at 3000rpm for 5 min until
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22 82 there was no milky interface with white precipitate, and decolorized with macroporous
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83 resin (Chen et al., 2019a).
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28 84 2.3. Characterization of BFP
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85 2.3.1. BFP monosaccharide component measurement
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33 86 The sample was measured by high-performance liquid chromatography (LC-
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35 87 20AD); an appropriate amount of sample was dissolved in a sample bottle, 0.6 mol/L
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37 88 NaOH and 0.4 mol/L PMP-methanol were added, and the reaction was carried out at
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39 89 70°C for 1h. Cooled in cold water for 10min; 500 μL of 0.3mol/L HCl was added to
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41 90 neutralize, and then chloroform was added and vortexed for 1min, centrifuged at
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43 91 3000r/min for 10min, and the supernatant was carefully taken and extracted three times.
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45 92 The chromatographic column model is Xtimate C18 4.6*200mm 5um, the column
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47 93 temperature is 30°C, the flow rate is 1.0ml/min, the detection wavelength is 250nm,
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49 94 and the mobile phase is 0.05M potassium dihydrogen phosphate solution (adjusted to
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51 95 pH 6.70 with sodium hydroxide solution)-acetonitrile=83-17 (Hu et al., 2021; Lo et al.,
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53 96 2011).
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55 97 2.3.2. Measurement of molecular weight of BFP
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57 98 The molecular weight and distribution of BFP were determined by gel permeation
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59 99 chromatography-multi-angle laser light scattering (GPC-MALLS). A 1.00mg/mL BFP
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 100 aqueous solution was prepared with the mobile phase (0.20mol/L NaCl solution). After
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101 centrifugation (4°C, 8000rpm, 10min), the sample solution was passed through a
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102 0.22um microporous membrane. The detection conditions were: Waters high-
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7 103 performance liquid chromatography system (Alliances 2695, Waters, USA), equipped
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9 104 with a gel filtration column (8.0×300.00mm, OHpak SB-806HQ, Japan, Shodex, Japan);
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11 105 flow rate was 0.40mL/min, NaCl solution was 0.20mol/L; injection volume was 200μL.
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13 106 The HPLC system was also equipped with a MALLS photometer (Dawn Heleos, Wyatt),
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15 107 a refractive index (RI) detector (Optilab rEX, Wyatt Technology Company, USA), and
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17 108 a multi-angle laser light scattering (LS) photometer (Dawn Heleos, Wyatt Technology
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19 109 Company, USA). During the chromatographic separation process, the absorbance of the
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21 110 was measured using MALLS-RI-LS detectors (Li et al., 2022).
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23 111 2.3.3. Water contact angle of BFP
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25 112 Contact angle (Powereach, JC2000C1) was used to test the hydrophilicity of BFP
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27 113 film, and pure water was suspended on the sample in a dry environment. The angle
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29 114 between the droplet and BFP film was calculated through the fitting of the instrument,
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31 115 and the hydrophilicity of the material was judged by the size of the angle (Zhu et al.,
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116 2021).
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36 117 2.3.4. Measurement of BFP interfacial tension
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38 118 The interfacial tension of BFP solution was measured by a liquid drop tensiometer
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40 119 (Teclis, France). The interfacial tension of BFP adsorbed at the oil-water interface at a
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42 120 specific adsorption time (t) was determined by the axisymmetric drop method. A U-
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44 121 shaped syringe containing MCT oil was immersed in a quartz tank containing different
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46 122 concentrations of BFP (0.10wt~0.50wt%) aqueous solution (12 mL). A 10 μL droplet
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48 123 of MCT oil was formed at the tip of the U-shaped syringe, and the shape of the droplet
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50 124 and the change of interfacial tension with time were continuously recorded by a camera.
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52 125 The entire detection process was carried out at room temperature (Chen et al., 2021;
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54 126 Han et al., 2020).
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56 127 2.3.5. Fourier-Transform Infrared (FTIR)
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58 128 The structure of BFP was determined by FT-IR. The dried sample was evenly
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60 129 mixed with KBr powder (1:100) for tablet pressing. Scanning was performed using
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 130 Nicolet iS10 Fourier transform infrared spectrometer in the wave range of 4000cm-1-
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131 400cm-1 (Zhang et al., 2022).
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6 132 2.4. Preparation of BFP emulsion
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133 The emulsion with an oil-water ratio of 6:4 was prepared by BFP solution
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11 134 (0.1wt%~2.5wt%) and MCT using phosphate buffer solution (pH 7.0). The mixture was
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13 135 pre-sheared (FA25 Fluko China) for 30 seconds at 10000 rpm using a high-speed
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15 136 shearing machine, and then homogenized for three minutes at 16000 rpm.
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17 137 The emulsions with 1.0wt% BFP and different oil-water ratios (4:6, 5:5, 6:4 and
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19 138 7:3) were prepared by BFP solution and MCT using phosphate buffer (pH 7.0). The
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21 139 mixture was pre-sheared for 30 seconds at 10000 rpm using a high-speed shearing
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23 140 machine, and then homogenized for three minutes at 16000 rpm.
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26 141 2.5. Characterization of BFP emulsions
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29 142 2.5.1. Microscope observation of BFP emulsion
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31 143 The droplet morphology of emulsion was observed and photographed with an
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33 144 optical microscope. 5 μL emulsion was absorbed and dropped on a glass slide, covered
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35 145 with a cover glass slide, and observed with a polarizing microscope (Nikon).
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37 146 2.5.2. Fluorescence microscope observation of BFP emulsion
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39 147 The microstructure of the emulsion was observed by fluorescence microscope
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148 (BZ-X800E). The prepared emulsion was diluted, 20 μL nile red (2mg/ml) solution was
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149 added into 1ml emulsion sample, shaken and mixed well, and the oil droplets were
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46 150 stained. A small amount of dyed emulsion was prepared on a glass slide, and observed
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48 151 at the wavelength of 488nm. All the above operations were performed under dark
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50 152 conditions.
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52 153 2.5.3. Zeta potential measurement of BFP emulsions
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54 154 The potential of emulsion droplets was measured using Zetasizer Nano-ZS
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56 155 (Malvern Instruments, UK). Before the test, the sample was shaken evenly, and 50μL
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58 156 emulsion was diluted in 4950 μL 10mM phosphate buffer solution to prevent scattering
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60 157 effect. The sample pool was a standard capillary electric pool (The light scattering angle
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 158 was 17° laser wavelength 633nm, He/Ne gas laser). The sample was measured at 25℃
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159 for three times in parallel.
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5 160 2.5.4. Average particle size(d32) and particle size distribution (PSD) of BFP
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7 161 emulsions
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9 162 The particle size of the emulsion was measured using static light scattering
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11 163 (Mastersizer2000, Malvern Instruments, UK), and phosphate buffer solution with pH=7
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13 164 was used as the dispersion medium. Before measurement, the sample was shaken
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15 165 evenly and added to the dispersion medium drop by drop until the signal meet the test
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17 166 requirements. The refractive index of the dispersed phase and the continuous phase
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19 167 were 1.475 and 1.300, respectively, and the pump speed was 2000r/min. Each sample
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21 168 was measured three times and averaged. The average particle size of the emulsion was
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23 169 expressed as d32.
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25 170 2.5.5. Determination of rheological properties of BFP emulsions
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27 171 The rheological behavior of oil-water interface of emulsion samples were
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29 172 measured, using a rheometer (HAAKE Rheostress6000, Scientific, USA), and a parallel
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31 173 plate with a diameter of 35.00 mm was selected as the measurement accessory with the
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174 gap set to 1.00 mm. The shear rate ranges was from 0.10 to 1000s⁻ ¹. Through dynamic
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175 stress scanning from 0.10 to 100Pa at a constant frequency of 0.1Hz, the linear
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38 176 viscoelastic region was determined, which was to facilitate the subsequent dynamic
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40 177 viscoelastic test. For the frequency sweep mode, the energy storage modulus (G') and
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42 178 loss modulus (G") of emulsions were measured in the frequency range from 0.1 to 10Hz
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44 179 at a stress of 0.5Pa. The 1.00~3.00mLemulsion sample to be measured was first
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46 180 uniformly laid on the sample table, and then the height of the rotor and the sample table
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48 181 was adjusted so that the rotor just touched the sample surface.
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51 182 2.6. Influence of environmental factors on the stability of emulsion
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54 183 2.6.1. Effect of heat treatment on emulsifying property of BFP emulsion
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56 184 BFP emulsion of 1.0wt% with an oil-water ratio of 6:4 was prepared and heated
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58 185 in a water bath at 60, 70, 80, 90℃ for 30 min, respectively. After cooling to room
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60 186 temperature, the emulsions were placed in a constant temperature incubator at 25°C for
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 187 12h. The mean particle size, particle size distribution and zeta-potential of the emulsion
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188 samples were measured, and the macroscopic changes were observed by polarizing
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189 microscope.
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7 190 2.6.2. Effect of pH on emulsifying property of BFP emulsion
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9 191 1.0wt% BFP emulsion with an oil-water ratio of 6:4 was prepared and divided into
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11 192 different vials. Then 1.00mol/L of HCl or NaOH solution was added to the emulsion
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13 193 system to adjust the pH value between 2.0 and 9.0. The emulsions were placed in a
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15 194 constant temperature incubator at 25℃ for 12h. The mean particle size, particle size
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17 195 distribution and zeta-potential of the emulsion samples were measured, and the macro
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19 196 changes were observed by polarizing microscope.
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21 197 2.6.3. Effect of ions on emulsifying properties of BFP emulsion
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23 198 After preparation 1.0wt% BFP emulsion with an oil-water ratio of 6:4, the BFP
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25 199 emulsion was divided into different bottles, and NaCl and CaCl₂ were added so that
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27 200 the content of NaCl in the final emulsion system was 0.100~0.500mol/L and the CaCl₂
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29 201 was 0.010~0.050mol/L, respectively. The emulsions were placed in a constant
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31 202 temperature incubator at 25℃ for 12h. The mean particle size, particle size distribution
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33
34
203 and zeta potential of the emulsion samples were measured, and the macro changes were
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204 observed by polarizing microscope.
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39 205 2.7. Simulation of in vitro digestion of curcumin-coated BFP emulsion
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42 206 Curcumin was added to the oil phase (MCT) and stirred magnetically for 12h to
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44 207 ensure the maximum solubility. Undissolved curcumin was removed by centrifuging at
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46 208 8000 rpm. A gastrointestinal digestion simulation model was constructed according to
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48 209 the method (Wooster et al., 2014) with some modification. Preparation of the simulated
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50 210 gastric acid: 2 g NaCl and 7 mL HCl (1 mol/L) were added to 1 L of deionized water.
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52 211 The final pH value of the digestive juice was adjusted to 2 with HCL (0.1 mol/L).
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54 212 Preparation of the simulated intestinal juice: 6.8 g KH2PO4 was added to 500 mL of
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56 213 deionized water and the pH value of the digestive juice was adjusted to 6.5 with NaOH
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58 214 (0.1 mol/L), then the solution was diluted to 1 L with deionized water. 1g sample was
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60 215 used for digestion experiment. After pH adjustment to 2.0 with HCl (0.1 mol/L), pepsin
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 216 (4000 U/mL) was added and digested at 50 rpm and 37 °C for 120 minutes. The pH
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217 value was adjusted to 6.5 with sodium acetate buffer and intestinal digestive juice was
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218 added after simulating gastric digestion for 120 minutes. BFP emulsion with a
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7 219 concentration of 2.0wt% was prepared using MCT dissolved with curcumin as the oil
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9 220 phase. 0.1mL BFP emulsion [2.0wt%BFP+MCT (cur)] was mixed with 3.9ml methanol,
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11 221 centrifuged, and the absorbance of the supernatant at 425nm was measured (Multiskan
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13 222 SkyHigh, Thermo Scientific, USA). Gastric juice was added to the emulsion in a ratio
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15 223 of 1:1 and digested at 37°C for 2 hours at a constant stirring speed of 50 rpm. 0.1ml of
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17 224 the mixed liquid after gastric digestion was mixed with 3.9ml methanol, centrifuged
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19 225 and the absorbance at 425nm was measured. Intestinal digestive juices in a ratio of 1:1
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21 226 was added and digested at 37°C for 2 hours at a constant stirring speed of 50 rpm. 0.1ml
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23 227 of mixed liquid after intestinal digestion was mixed with 3.9ml methanol, centrifuged
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25 228 and the absorbance at 425nm was measured. The protective effect of BFP emulsion on
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27 229 curcumin was determined by comparing the absorption value of BFP emulsion with Ga
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29 230 emulsion of the same concentration at different stages of digestion. The calculation
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31 231 formula of the bioavailability, loading rate and retention rate of curcumin is:
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33 𝐶
34 232 Bioavailability = 𝐶𝑖𝑑 × 100
𝑙𝑎
35
𝐶
36
37
233 Loading rate = 𝐶𝑙𝑎 × 100
𝑡𝑎
38
𝐶𝑐𝑎
39 234 Retention rate = × 100
40 𝐶𝑙𝑎
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42 235 Here, Cid represents the light absorption value at the end of intestinal digestion. Cla
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44 236 represents the absorption value of curcumin loaded with emulsion. Cta represents the
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46 237 light absorption value of curcumin loaded by MCT. Cca represents the light absorption
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48 238 value of curcumin in the stomach stage and intestinal stage.
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50
51 239 2.8. Statistic analysis
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54 240 All data was analyzed by Origin 21. A nonrepetitive two-factor analysis of
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56 241 variance (ANOVA) was performed to determine the statistical significance of each
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58 242 group of data obtained from three independent assays on each sample.
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 1 243 3.Results and Discussion
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5 244 3.1. Characterization of BFP
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7 245 3.1.1. Monosaccharide component analysis
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246 The monosaccharides of buckwheat flower polysaccharide were mannose, ribose,
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247 rhamnose, glucose, galactose, arabinose and fucose. The molar ratio was
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14 248 1.82:1.54:6.68:8.73:11.21:9.41:1.00
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16 249 3.1.2. Molecular weight
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18 250 The polysaccharide with low molecular weight and average molecular weight of
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20 251 BFP was 2.421×105 Da, which could be quickly adsorbed on the oil-water interface and
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22 252 help to reduce the interfacial free energy. The higher molecules of the polysaccharide
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24 253 prevented the instability of the emulsion and made it more stable in the emulsion (Li et
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26 254 al., 2018).
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28 255 3.1.3. FT-IR analysis
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30 256 The FT-IR results were shown in Fig.1A. There was a distinct absorption peak
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32 257 near 3274 cm-1, most probably caused by the contraction vibration of hydroxyl groups
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34 258 within or between polysaccharide molecules, which was the characteristic peak of
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36 259 hydroxyl groups on sugar rings (F. Chen et al., 2019). The absorption peak at 2934cm-1
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38 260 was the C-H stretching vibration absorption peak, the absorption peak at 1634cm-1 was
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40 261 the C-O asymmetric vibration peak, and the absorption peaks at 1544cm-1 and 1416cm-1
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42
43
262 were the C-H stretching vibration peaks (Sebeia et al., 2019). A distinct absorption peak
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45 263 appeared at 1236cm-1, which might be due to the stretching and bending vibrations of
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47 264 CH3 at C-H and C-O. The absorption peak near 1015cm-1 might be caused by the C-O-C
48
49 265 or C-OH stretching vibration of the pyranose ring (Zhang et al., 2022). The absorption
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51 266 peak at 588cm-1 might be caused by the contraction vibration of the D-ribose glucose
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53 267 ring (Wiercigroch et al., 2017). It could be inferred that the sugar ring of buckwheat
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55 268 flower polysaccharide was β-D-pyranose glucose ring (He et al., 2016).
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57 269 3.1.4. Water contact angle
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59 270 The contact angle (θO/W) can be used as a direct way to evaluate the surface
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 271 wettability of the sample, which plays a crucial role in the adsorption and stabilization
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272 of particles at the oil/water interface (Wang et al., 2015). Studies have shown that when
4
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273 the water contact angle is less than 90°, liquid is more likely to wet solid materials,
6
7 274 indicating that the solid surface is hydrophilic; when the water contact angle is greater
8
9 275 than 90°, the liquid is not easy to wet solid materials, indicating that the solid surface
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11 276 is hydrophobic (Gu et al., 2016). In this study, the water contact angle of BFP tablet
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13 277 was 32.7°and 39.8° (Fig.1C), which indicated that BFP was hydrophilic and had
14
15 278 excellent advantages in adsorption at the water interface, making it easier to make
16
17 279 water-in-oil emulsions.
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19
20 280 3.2. Interfacial tension of BFP
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23 281 At the beginning of the measurement, the interfacial tension decreased rapidly
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25 282 (Fig.1B), indicating that BFP was adsorbed at the oil-water interface. As the
26
27 283 concentration of BFP increased, the interfacial tension became lower. It decreased from
28
29 284 17.2 mN/m to 14.1 mN/m, and the decrease in interfacial tension could initially
30
31 285 determine the emulsifying ability of BFP. Ga, as a natural plant polysaccharide, is
32
33 286 widely used in the food industry due to its excellent emulsifying properties (Cui et al.,
34
35 287 2021; Shao et al., 2020; Williams & Phillips, 2009). As shown in Fig.1B, the interfacial
36
37
288 tension of 0.2% BFP (16.2mN/m) was significantly lower than that of commercial
38
39
40
289 emulsifier Ga (18.5mN/m), which displayed the potential of BFP as a natural emulsifier.
41
42
43 290 3.3. Effect of different polysaccharide ratio on emulsion
44
45
46 291 The addition of different proportions of natural polysaccharides also affects the
47
48 292 stability of the emulsion. In the emulsion system, the stability of the emulsion can be
49
50 293 improved by increasing the concentration of the emulsifier, but excessive emulsifier
51
52 294 can also cause food safety problems. This rule must also be followed when exploring
53
54 295 the emulsifying properties of BFP (Y. Hu et al., 2020; Mayo-Prieto et al.,2022). The
55
56 296 particle size distribution, d32 and zeta potential of BFP emulsions with different BFP
57
58 297 concentrations (0.1wt%, 0.5wt%, 1.0wt%, 2.0wt%, 2.5wt%, pH 7.0) were shown in
59
60 298 Fig.2A. As the concentration of BFP increased, the d32 of the emulsion decreased from
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62
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 299 60.0μm to 8.3μm. The d32 of emulsions with BFP concentrations of 0.1wt% and 0.5wt%
1
2
3
300 was larger, which might because low-concentration polysaccharides could not
4
5
301 completely wrap oil droplets, resulting in the formation of larger droplet aggregates (Xu
6
7 302 et al., 2016). When the BFP concentration was high (above 1.0wt%), the particle size
8
9 303 distribution was more uniform and the d32 of the emulsion was low, indicating that the
10
11 304 droplets were completely wrapped by polysaccharides. The absolute potential value of
12
13 305 emulsion droplets reflects the surface charge density and repulsive force between
14
15 306 droplets. The stability of emulsion is positively correlated with the charge on the surface
16
17 307 of droplets (Chen et al., 2019b). Zeta potential is an indicator to measure the stability
18
19 308 of emulsion caused by electrostatic repulsion. The smaller the particle size, the higher
20
21 309 the absolute value of zeta potential (Nakauma et al., 2008). When BFP concentration
22
23 310 was greater than 1.0wt%, as BFP concentration increased, the absolute value of zeta
24
25 311 potential increased, indicating that emulsion stability was enhanced (McClements &
26
27 312 Gumus, 2016). At the same time, BFP emulsion with a concentration of 1.0wt% had
28
29 313 the same emulsifying ability as Ga emulsion with a concentration of 2.0wt%, further
30
31 314 demonstrating the excellence of BFP as a surfactant.
32
33
34
315 From the particle size distribution chart (Fig.2B), it could be seen that BFP
35
36
316 emulsion with a concentration of 1.0wt% had a more uniform particle size distribution
37
38 317 and was unimodal while other concentrations were bimodal. Combining Fig.2A and
39
40 318 considering the food safety requirements, that food additives should be as little as
41
42 319 possible, subsequent experiments will be conducted using a ratio of 1.0wt% BFP.
43
44
45 320 3.4. Effect of oil phase ratio on emulsion
46
47
48 321 The oil phase ratio had little effect on the average particle size (Fig.2C), but the
49
50 322 zeta potential value of the BFP emulsion with an oil phase ratio of 6:4 was the highest,
51
52 323 and the particle size distribution chart (Fig.2D) also showed that the particle size
53
54 324 distribution was more uniform with a unimodal distribution. Also, the droplet size of
55
56 325 6:4 oil-water ratio under fluorescence microscopy was uniform, completely stained by
57
58 326 nile red (Fig.3A), indicating that the emulsion type was a water-in-oil emulsion.
59
60 327 Therefore, emulsions with oil-water ratio of 6:4 and 1.0wt% BFP were used in the
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 328 subsequent experiments.
1
2
3
4
329 3.5. Effect of heat treatment on the stability of BFP emulsion
5
6
7
330 The stability of emulsified food during manufacturing, transportation, and storage
8
9
331 is important, and thermal stability is an important indicator for evaluating this stability.
10
11 332 Since many foods undergo high-temperature sterilization and other procedures during
12
13 333 production and processing, and are sometimes stored in high-temperature environments,
14
15 334 good thermal stability is very beneficial for the application of emulsified foods (J.
16
17 335 O’Mahony et al., 2017). The thermal stability of BFP emulsion with an oil-water ratio
18
19 336 of 6:4 and a BFP emulsifier content of 1.0wt% was explored. When heated, the droplets
20
21 337 of the emulsion did not change significantly under the microscope (Fig.3B) and were
22
23 338 uniformly spherical without aggregation. However, as the treatment temperature
24
25 339 increased, the particle size of the emulsion tended to increase (Fig.4A). The average
26
27 340 particle size of BFP emulsion after heat treatment increased from 9.21μm (25℃) to
28
29 341 10.84μm (60℃), 11.95μm (70℃), 12.12μm (80℃), and 13.83μm (90℃). Heat treatment
30
31 342 might have some impact on the particle size of the emulsion (Fig.4B). From the particle
32
33 343 size distribution chart, it could be seen that the peak shape had a tendency to shift to the
34
35 344 right. But the zeta potential of BFP emulsion after heat treatment (Fig.4A) changed little.
36
37
345 The results indicated that BFP emulsion had good heat resistance.
38
39
40
346 Rheological properties are an important indicator for evaluating the stability of
41
42 347 polymers and a method for understanding the complex structure of solvents. At the
43
44 348 same time, the stability was characterized by measuring the apparent viscosity and
45
46 349 frequency scanning test of BFP after heat treatment. The apparent viscosity of BFP
47
48 350 emulsion after heat treatment decreased with increasing shear rate, showing shear
49
50 351 thinning phenomenon, which may be due to the destruction of internal structure by
51
52 352 shear force. As the treatment temperature increased, the apparent viscosity of BFP also
53
54 353 showed an increasing trend (Fig.5A). Generally speaking, the higher the apparent
55
56 354 viscosity, the higher the stability of the emulsion (Ge et al., 2021). It is shown that BFP
57
58 355 emulsion after heat treatment has better stability. The changes in storage modulus (G')
59
60 356 and loss modulus (G") under fixed stress (0.5Pa) at 1~5Hz were also studied for BFP
61
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 357 emulsion after heat treatment. As shown in Fig.5B, G' was greater than G", indicating
1
2
3
358 that it was still mainly elastic after heat treatment and exhibit gel-like properties. Both
4
5
359 G' and G" showed an increasing trend, which might be due to the increase in
6
7 360 intermolecular forces as the particle size of the emulsion increased after heat treatment,
8
9 361 resulting in enhanced viscoelasticity of the emulsion (Y.-T. Xu et al., 2018). Thermal
10
11 362 stability indicated that BFP emulsion had excellent heat-resistant properties.
12
13
14 363 3.6. Effect of pH on emulsifying property of BFP emulsion
15
16
17 364 Microscopic observation of 1.0wt% BFP emulsion under different pH values
18
19 365 (Fig.3B) showed that acidic environment had little effect on emulsion particle size, and
20
21 366 the particle size of the emulsion had hardly changed. When the pH was 8 and 9, the
22
23 367 particle size of the emulsion was significantly larger. It can be seen from the result that
24
25 368 (Fig.6A), when the pH was 2, 4, 6, and 7, there was no significant change in the particle
26
27 369 size. That means, acidic environments hardly affected BFP emulsion. However, when
28
29 370 the pH value increased to 8 and 9, the particle size of the emulsion increased from
30
31 371 18.20μm to 25.96μm; the zeta potential of the emulsion after treatment showed a
32
33 372 downward trend. From the result of Fig.6B, it could be seen that the peak shape had a
34
35 373 clear trend of enlargement, which may be due to the increase in anions in the emulsion
36
37
374 adsorbed on cations of polysaccharides, thereby reducing intermolecular electrostatic
38
39
40
375 repulsion and adsorbing larger droplets (Xu et al., 2017).
41
42 376 The apparent viscosity and viscoelasticity of BFP emulsion under the pH of 2, 4,
43
44 377 and 6 all showed an increasing trend (Fig.7), which may be due to the conversion of -
45
46 378 NH2 group to -NH3+ under acidic conditions, increasing the intermolecular electrostatic
47
48 379 interactions and reducing the possibility of droplet aggregation. Under alkaline
49
50 380 conditions, the apparent viscosity and viscoelasticity showed a decreasing trend (Fig.7),
51
52 381 which may be due to the increase of OH- reducing the intermolecular electrostatic
53
54 382 repulsion and weakening the rheological properties of the emulsion, which was
55
56 383 consistent with the previous inferences.
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 1 384 3.7. Effect of ions on emulsifying properties of BFP emulsion
2
3
4 385 The microscopic observation of the effect of ions on the emulsion was shown in
5
6 386 Fig.3B. It was shown that different concentrations of Na+ and Ca2+ had little effect on
7
8 387 the emulsion droplets. As was shown in Fig.8, the average particle size of BFP emulsion
9
10 388 fluctuated around 10 μm, displaying unimodal without aggregation under different ion
11
12 389 and different concentration conditions, and the fluctuation of zeta potential was not
13
14 390 obvious.
15
16 391 Similarly, there was an increasing trend in the apparent viscosity and
17
18 392 viscoelasticity of BFP emulsion than that of the control emulsion without treated ions
19
20 393 (Fig.9).
21
22 394 The addition of ions increased the adsorption density of polysaccharides at the
23
24
395 interface. The change in ion strength in the aqueous phase promoted the interaction
25
26
27
396 between hydrophobic chains of polysaccharides, while cations connected the
28
29 397 hydrophilic polysaccharides, thereby reducing interfacial tension and enhancing the
30
31 398 rheological properties of the interfacial membrane (Chen et al., 2023).
32
33
34 399 3.8. Simulation digestion of curcumin-coated 1.0wt%BFP emulsion
35
36
37 400 Curcumin is a highly lipophilic compound, which limits its absorption in the
38
39 401 human body. It has a relatively high metabolic degradation rate during physiological
40
41 402 transport, and the final metabolites are inactive and rapidly eliminated from the body,
42
43 403 reducing the bioavailability of curcumin (Hales et al., 2020). It was shown from Table.1
44
45 404 that the bioavailability of curcumin in the human body was only 7.10%, which was
46
47 405 consistent with the easy degradation of curcumin in biological transport. However, BFP
48
49 406 emulsion with 1.0wt% and 60wt% oil phase had a good protective effect on curcumin,
50
51 407 increasing the retention rate of curcumin from 40.24% to 64.47% during the gastric
52
53 408 digestion stage and from 17.64% to 41.97% during the intestinal digestion stage,
54
55 409 increasing the bioavailability of curcumin from 7.10% to 27.05%. Compared with Ga
56
57 410 emulsion of the same concentration, all indicators of BFP emulsion are higher than Ga.
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 1 411 4.Conclusions
2
3
4
5 412 At present, most of the emulsifiers used in food are synthetic emulsifiers. With the
6
7 413 increasing attention to healthy food, the application of natural emulsifiers has recently
8
9 414 attracted much attention. The buckwheat flower polysaccharide (BFP) from discarded
10
11 415 buckwheat flowers was prepared as a novel source of emulsifier, and the properties and
12
13 416 stability of high oil phase of BFP emulsions were characterized in this study. When the
14
15 417 oil phase was 60.0 wt%, the emulsifying stability of the emulsion with 1.0 wt% BFP
16
17 418 was the strongest. At this concentration, BFP emulsion had good heat resistance, ion
18
19 419 resistance, and acid resistance, and the average particle size always fluctuated around
20
21 420 10μm. However, under alkaline conditions, due to the influence of static electricity, the
22
23 421 particle size become larger and had a certain impact on the stability of the emulsion. In
24
25 422 the simulated digestion experiment, BFP emulsion had a good protective effect on
26
27 423 curcumin, significantly increasing its retention rate during gastric and intestinal
28
29 424 digestion stages. This study indicates that BFP can be used as a new natural emulsifier,
30
31
32
425 not only providing new ideas for the recycling of buckwheat flower resources, but also
33
34
426 for the future development of BFP in the fields of food, medicine, cosmetics, etc.
35
36 427 However, the emulsification mechanism of BFP remains to be further explored.
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38 428
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 1 429 Acknowledgements
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5 430 This research was financially supported by Hubei Natural Science Foundation
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7 431 (2022CFB476).
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25 562 https://doi.org/10.1533/9781845695873.252
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27 563 Wooster, T. J., Day, L., Xu, M., Golding, M., Oiseth, S., Keogh, J. & Clifton, P. (2014).
28
29 564 Impact of different biopolymer networks on the digestion of gastric structured
30
31 565 emulsions. Food Hydrocolloids, 36, 102–114.
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33
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566 https://doi.org/10.1016/j.foodhyd.2013.09.009
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36
567 Xu, X., Zhong, J., Chen, J., Liu, C., Luo, L., Luo, S., Wu, L. & McClements, D. J.
37
38 568 (2016). Effectiveness of partially hydrolyzed rice glutelin as a food emulsifier:
39
40 569 Comparison to whey protein. Food Chemistry, 213, 700–707.
41
42 570 https://doi.org/10.1016/j.foodchem.2016.07.047
43
44 571 Xu, Y.-T., Tang, C.-H., Liu, T.-X. & Liu, R. (2018). Ovalbumin as an Outstanding
45
46 572 Pickering Nanostabilizer for High Internal Phase Emulsions. Journal of Agricultural
47
48 573 and Food Chemistry, 66(33), 8795–8804. https://doi.org/10.1021/acs.jafc.8b02183
49
50 574 Xu, X., Luo, L., Liu, C. & McClements, D. J. (2017). Utilization of anionic
51
52 575 polysaccharides to improve the stability of rice glutelin emulsions: Impact of
53
54 576 polysaccharide type, pH, salt, and temperature. Food Hydrocolloids, 64, 112–122.
55
56 577 https://doi.org/10.1016/j.foodhyd.2016.11.005
57
58 578 Zhao, J., Jiang, L., Tang, X., Peng, L., Li, X., Zhao, G. & Zhong, L. (2018). Chemical
59
60 579 Composition, Antimicrobial and Antioxidant Activities of the Flower Volatile Oils
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 580 of Fagopyrum esculentum, Fagopyrum tataricum and Fagopyrum Cymosum.
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581 Molecules, 23(1), 182. https://doi.org/10.3390/molecules23010182
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582 Zhang, Y., Li, K., Chen, M., Fang, S., Zhen, D., Cao, J., Wu, Z. & Zhang, K. (2022). A
6
7 583 novel polysaccharide prepared from Chrysanthemum morifolium cv. Fubaiju tea and
8
9 584 its emulsifying properties. International Journal of Food Science & Technology,
10
11 585 57(6), 3385–3399. https://doi.org/10.1111/ijfs.15658
12
13 586 Zhu, X., Chen, J., Hu, Y., Zhang, N., Fu, Y. & Chen, X. (2021). Tuning complexation
14
15 587 of carboxymethyl cellulose/ cationic chitosan to stabilize Pickering emulsion for
16
17 588 curcumin encapsulation. Food Hydrocolloids, 110, 106135.
18
19 589 https://doi.org/10.1016/j.foodhyd.2020.106135
20
21 590 Zhu, LiuYing; Li, Juan; Wei, Changhao; LUO, TING; Deng, Zeyuan; Ya, Weifan;
22
23 591 Zheng, Liufeng (2020). Polysaccharide from Fagopyrum esculentum Moench bee
24
25 592 pollen alleviates microbiota dysbiosis to improve intestinal barrier function in
26
27 593 antibiotic-treated mice. Food & Function, 10.1039. D0FO01948H–.
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29 594 https:/doi:10.1039/D0FO01948H
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31 595
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 1 596 Figure captions
2
3
4
5 597 Fig.1.Physicochemical properties of BFP, Fourier infrared spectrum (A), Interfacial
6
7 598 tension(B), Water contact angle (C)
8
9 599 Fig.2.Average particle size, zeta potential and particle size distribution of different
10
11 600 polysaccharide proportions（A,B) and different oil phase ratio (C,D)
12
13 601 Fig.3.Fluorescence microscopy of BFP emulsion (A) and polarization microscopy
14
15 602 under different conditions (pH, temperature, ion)(B)
16
17 603 Fig.4.1.0wt% BFP (60% oil phase) emulsion heated at different temperatures (40 to
18
19 604 90°C) d32, zeta-potential (A) and PSD (B)
20
21 605 Fig.5.Viscosity curves (A) and frequency-modulus curves (B) of 1.0wt%BFP (60% oil
22
23 606 phase) emulsion at different temperatures (40~90℃). The experiment was conducted
24
25 607 at 25°C.
26
27 608 Fig.6.1.0wt% BFP (60% oil phase) emulsion at different pH (pH2~pH9) d32, zeta
28
29 609 potential (A) and PSD (B)
30
31
32
610 Fig.7.Viscosity curves (A) and frequency-modulus curves (B) of 1.0wt%BFP (60% oil
33
34
611 phase) emulsion at different pH (pH2~pH9). The experiment was conducted at 25°C.
35
36 612 Fig.8.1.0wt% BFP (60% oil phase) emulsion at different ions added (0.1~0.5mo
37 + 2+
38 613 l/L Na and 0.01~0.05mol/L Ca ) d32, zeta potential and PSD.
39
40 614 Fig.9. Viscosity curves and frequency-modulus curves of 1.0wt%BFP (60% oil phase)
41
42 615 emulsion at different ions added (0.1~0.5mol/L Na + and 0.01~0.05mol/L Ca2+). The
43
44 616 experiment was conducted at 25°C.
45
46 617
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 618 Fig.1.
1
2
3
619
4
5
6
7
8
9
10
11
12
13
14
15
16 24
17

A B
18 0.1wt%BFP
0.2wt%BFP

Interfacial tension (mN/m)

19 22 0.3wt%BFP
0.4wt%BFP
20 0.5wt%BFP
0.2wt%Ga
21 20
22
23
18
24
25
26 16
27
28
14
29 0 500 1000 1500 2000 2500 3000 3500
30 Time (s)
31
32
33
34
C
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 620 Fig.2.
1
2
3
621
4
5
6
7
8 80
Meam particle size 14 2.5wt%
9
10 A Zeta potential -40
B 12
2.0wt%
1.0wt%
0.5wt%
0.1wt%
11 -38 2.0wt%GA
Meam particle size (μm)

60
12

Zeta potential (mV)

10
13

Volume (%)
-36
14 8
15 40
-34 6
16
17 4
-32
18 20
19 2
20 -30

21 0
22 0 -28
2.5 2.0 1.0 0.5 0.1 2.0GA 0.1 1 10 100 1000 10000
23
24 Concentration (wt%) Partical size (μm)
-37 16
25 Meam particle size
26
27
C Zeta
-36 D 14
4:6
5:5
10 6:4
28
Meam particle size (μm)

12 7:3
-35
29
10
30
Zeta (mV)

volume (%)

-34
31 8
32
-33
33 5 6
34
-32 4
35
36 2
-31
37
38 0
0 -30
39 4:6 5:5 6:4 7:3 0.01 0.1 1 10 100
40 Proportion（ O/W) Partical size (μm)
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 622 Fig.3.
1
2
3
623
4
5
6
7
A
8
9
10
11
12
13
14
15
16
17
B 25℃ 60℃ 70℃ 80℃ 90℃
18
19
20
21
22
23
24
25
26
27
pH2 pH4 pH6 pH7 pH8 pH9
28
29
30
31
32
33
34
35
36
37 0.01M Ca2+ 0.02M Ca2+ 0.03M Ca2+ 0.04M Ca2+ 0.05M Ca2+
38
39
40
41
42
43
44
45
46
47 0.10M Na+ 0.20M Na+ 0.30M Na+ 0.40M Na+ 0.50M Na+
48
49
50
51
52
53
54
55
56
57 624
58
59 625
60
61
62
63
64
65
 626 Fig.4.
1
2
3
627
4
5
628
6
7 629
8
9 630
10
11 631
12
13 632
14
15
16 633
17
18 634
19
20 635
21 -40
25℃
A 22
23
Mean particle size
Zeta potential B
16
60℃
15
24 14 70℃
25 -38
80℃
Mean particle size (μm)

26 12
90℃
Zeta potential (mV)

27
volume (%)

10
28 -36
10
29
8
30
31 -34 6
32
33
5 4
34
35 -32
2
36
37 0
38
0 -30
39 25 60 70 80 90 0.1 1 10 100 1000
40 Temperature (℃ ) Partical size (μm)
41
42 636
43
44 637
45
46 638
47
48 639
49
50 640
51
52
641
53
54
55
642
56
57 643
58
59
60
61
62
63
64
65
 644 Fig.5.
1
2
3
645
4
5
646
6
7 647
8
9 648
10
11 649
12
13
14
650
15
16 651
17
18 652
19
20 653
21
22 654
23
24 655
25
26 656
27
28
100000
29 25℃ 60℃ 70℃ 80℃ 90℃ G'
25℃ 25℃ 60℃ 70℃ 80℃ 90℃ G"
A 30
31
60℃ B
70℃ 100 100
32 80℃
33 90℃
34
10000
35
η (mPas)

36

G"(Pa)
G' (Pa)

37
38
39
401000
41
42 10 10
43
44
45
100
46 0.1 1 10 100 1000 1 2 3
47
48 ɣ̇ (1/s) f(Hz)
49
50
657
51
52 658
53
54 659
55
56 660
57
58 661
59
60
61
62
63
64
65
 662 Fig.6.
1
2
3
663
4
5
664
6
7 665
8
9 666
10
11 667
12
13
14
668
15
16 669
17
18 670
19
20
30
21 -36 16

A22
Mean particle size
Zeta potential B 14
pH2
pH4
23
25 -34 pH6
24
12 pH7
Mean particle size (μm)

25
Zeta potential (mV)

26 pH8
20 10
27 -32 pH9
volume (%)

28
8
29
15
30 -30
6
31
10
32
-28 4
33
34
5 2
35
36 -26
0
37
0
38 pH2 pH4 pH6 pH7 pH8 pH9 0.1 1 10 100 1000
39
40 pH Partical size (μm)
41 671
42
43
44
672
45
46
673
47
48 674
49
50 675
51
52 676
53
54 677
55
56 678
57
58 679
59
60 680
61
62
63
64
65
 681 Fig.7.
1
2
3
682
4
5
683
6
7 684
8
9 685
10
11 686
12
13 687
14
15 688
16
17 689
18 pH2 pH4 pH6 pH8 pH9 G'
100000
19 pH2 pH2 pH4 pH6 pH8 pH9 G"

A
20
21
pH4 B
pH6
22 pH7 100 100
23 pH8
24 pH9
10000
25
η (mPas)

26
G' (Pa)

G"(Pa)
27 10 10
28
29
30 1000
31
32
1 1
33
34
35 100
36 0.1 1 10 100 1000 0.1 1
37 ɣ̇ (1/s) f(Hz)
38
39 690
40
41 691
42
43 692
44
45 693
46
47 694
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 695 Fig.8.
1
2
3
696
4
5 -40 16

A 6
Mean particle size
Zeta potential B 14
0.1mol/L Na+
7 0.2mol/L Na+
108 -38 0.3mol/L Na+
12
Mean particle size (μm)

9 0.4mol/L Na+

Zeta potential (mV)

10
11 10 0.5mol/L Na+

volume (%)
-36 CK
12
8
13
14
5 -34 6
15
16
4
17
18 -32
2
19
20
0
21
0 -30
22 0.1 0.2 0.3 0.4 0.5 0.1 1 10 100 1000
23
24 Concentration (mol/L) Partical size (μm)
C 25
26
Mean particle size
Zeta potential -34
D 18
0.01mol/L Ca2+
16
27 0.02mol/L Ca2+
10
28 14 0.03mol/L Ca2+
Mean particle size (μm)

29 0.04mol/L Ca2+
Zeta potential (mV)

-32
30 12
0.05mol/L Ca2+
31
volume (%)

32 10 CK
33 -30
8
34
5
35 6
36 -28
37 4
38
39 2
40 -26
0
41
0
42 0.01 0.02 0.03 0.04 0.05 0.1 1 10 100 1000
43
44 Concentration (mol/L) Partical size (μm)
45
46
697
47
48
698
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 699 Fig.9.
1
2
3
700
4
5
701
6
7 702
8
9 703
10
11 0.01mol/L Ca2+ 0.02mol/L Ca2+ 0.03mol/L Ca2+ 0.04mol/L Ca2+ 0.05mol/L Ca2+ G'
100000 0.01mol/L Ca2+ 0.02mol/L Ca2+ 0.03mol/L Ca2+ 0.04mol/L Ca2+ 0.05mol/L Ca2+ G"
12
0.01mol/L Ca2+
13 0.02mol/L Ca2+
A
14
15
0.03mol/L Ca2+
0.04mol/L Ca2+
B 100 100

16 0.05mol/L Ca2+
17
10000 CK
18
η (mPas)

19

G"(Pa)
20 G' (Pa)
21 10 10
22
231000
24
25
26
27
28 100 1 1
29 0.1 1 10 100 1000 0.1 1
30 f(Hz)
ɣ̇ (1/s)
31
100000 0.10mol/L Na+ 0.20mol/L Na+ 0.30mol/L Na+ 0.40mol/L Na+ 0.50mol/L Na+ G'
32 + 0.10mol/L Na+ 0.20mol/L Na+ 0.30mol/L Na+ 0.40mol/L Na+ 0.50mol/L Na+ G"
0.10mol/L Na

C D
33 0.20mol/L Na+
34 0.30mol/L Na+
35 0.40mol/L Na+ 100 100
36 0.50mol/L Na+
10000
37 CK
38
η (mPas)

39
G' (Pa)

G"(Pa)
40
41
421000 10 10
43
44
45
46
47
48100 1 1
49 0.1 1 10 100 1000 0.1 1
50 ɣ̇ (1/s) f(Hz)
51
52 704
53
54 705
55
56 706
57
58 707
59
60 708
61
62
63
64
65
 709 Table .1.
1
2
3
710 The result of simulation digestion of curcumin-coated BFP emulsion
4
5
6 MCT BFP Ga
7
8
9 100.00% 89.71%±1.37% 88.09%±3.01%
10 Load rate
11
12
13 40.24%±1.24% 64.47%±2.46% 59.65%±1.79%
14 Gastric retention rate
15
16
17 17.64%±2.32% 41.97%±1.33% 39.24%±1.54%
18 Intestinal retention rate
19
20
21 7.10%±1.72% 27.05%±2.13% 23.40%±1.63%
22 Bioavailability
23
24
25 711
26
27 712
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65