Rutin-modified soybean protein isolate-pectin-genipin nanoparticle carrier

Abstract

The invention provides a rutin-modified soybean protein isolate-pectin-genipin nano-particle carrier and a preparation method of the rutin-modified soybean protein isolate-pectin-genipin nano-particle carrier. The preparation method of the nanoparticle carrier comprises the following steps: preparing a soybean protein isolate solution, a soybean protein isolate solution after pH shift, an aqueous solution containing rutin-modified soybean protein isolate and an aqueous solution containing rutin-modified soybean protein isolate-pectin; the aqueous solution contains rutin-modified soy isolate protein-pectin-genipin, the concentration of the genipin is 0.05 mg / mL, and the rutin-modified soy isolate protein-pectin-genipin nano-particle carrier is prepared from the rutin-modified soy isolate protein-pectin-genipin nano-particle carrier. The invention further provides application of the rutin-modified soy isolate protein-pectin-genipin nanoparticle carrier. The rutin-modified soy isolate protein-pectin-genipin nanoparticle carrier is used for sustained release of gastric drugs and targeted release of drugs in small intestines. According to the preparation method disclosed by the invention, a rutin-modified soybean protein isolate-polysaccharide nanoparticle carrier with a shell-core structure can be formed, gastric acid erosion can be effectively resisted, and the rutin-modified soybean protein isolate-polysaccharide nanoparticle carrier is delivered to small intestines in a targeted manner.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of food science and technology, and particularly relates to a preparation method of rutin-modified soybean protein isolate-pectin-gossypol nanoparticles carrier and application thereof. BACKGROUND

[0002] Rutin is the main biofunctional component of medicinal and edible materials Flos Sophorae and Sophora Fruit, and has many benefits such as anti-inflammatory, antioxidant, and anti-tumor effects. However, rutin belongs to the fourth class in the BCS drug classification system, has very low water solubility (<0.1 mg / mL), and poor membrane permeability, which affects its absorption in the body and leads to low oral availability (<2%), thereby severely limiting its application potential and resulting in almost no related food development on the market.

[0003] Currently, there are many studies on encapsulation delivery carriers for rutin, such as cyclodextrin inclusion compounds, phospholipid complexes, solid dispersions, liposomes, protein nanoparticles, and microcapsules. However, these methods are limited by inherent defects, such as poor stability of cyclodextrin inclusion compounds, easy rutin leakage; rutin gelatin or chitosan microspheres have large and uneven particle sizes, and need to add a crosslinking agent with safety risks; rutin liposomes have high biocompatibility and strong skin permeability, but have poor stability, and are prone to aggregation and flocculation due to the presence of oil in the system; polymeric micelles have high thermodynamic stability, but are prone to aggregation in the gastrointestinal tract, leading to burst release and subsequent degradation of rutin, and poor absorption effect; nanostructured lipid carriers use solid oil as the skeleton shell for coating active substances, which have good biocompatibility and safety with rutin, but have low encapsulation efficiency, are not stable in the body after administration, and have poor storage stability; and preparation of surface-modified composite encapsulation nanocrystals is difficult, has high cost, and is not conducive to large-scale production. Therefore, how to develop a rutin delivery carrier with good solubility characteristics, avoid the above problems as much as possible, improve the drug loading capacity of rutin encapsulation preparations, balance the release rate and release time, and improve the stability of the preparation itself is an urgent problem to be solved.

[0004] Because the human digestive system is an extremely complex system, including mouth, esophagus, stomach, small intestine and large intestine, the physicochemical environment (such as pH value, ion composition, enzyme and temperature) from mouth to large intestine is not the same, and the small intestinal epithelial cell as an absorption organ, the mucus barrier on its surface is a relatively hydrophobic gel network composed of highly glycosylated mucin, the delivery carrier needs to resist the erosion of gastric acid first, and also needs to avoid the charge adsorption effect and combine with the gastric mucosa, so as to enter the small intestine and penetrate and diffuse from the gastric mucosa mucus layer, so that the rutin can quickly cross the mucus barrier to reach the cell layer for absorption. However, the current technology does not conduct in-depth research in this regard, and the problem of the bioavailability of the delivered polyphenol is not well solved. Therefore, there is an urgent need to provide a delivery carrier capable of loading rutin, resisting gastric acid erosion, penetrating gastric mucosa, and a carrier preparation method for improving the water solubility and oral bioavailability of rutin. SUMMARY

[0005] The technical problem to be solved by the present application is to provide a preparation method of a rutin-modified soybean protein isolate-pectin-genepein nanoparticle carrier and its application, which can form a non-covalent good water-soluble delivery carrier with rutin and effectively improve the bioavailability of rutin, and on this basis, by compounding negatively charged pectin, a rutin-soy protein-polymer nanoparticle carrier with a "core-shell" structure can be formed, which can effectively resist gastric acid erosion and be targeted to the small intestine.

[0006] To solve the above technical problems, the technical scheme adopted by the present application is as follows: a preparation method of a rutin-modified soybean protein isolate-pectin-genepein nanoparticle carrier, which comprises the following steps: S1, dispersing soybean protein isolate powder in distilled water, stirring and keeping the pH value of the system at 7.0 to obtain a soybean protein isolate solution with a mass concentration of 10 mg / mL; S2, adjusting the pH value of the soybean protein isolate solution with a mass concentration of 10 mg / mL obtained in S1 to 12.0 with sodium hydroxide solution, stirring for 1 h, then adjusting the pH value to 7.0, and stirring for 20 min to obtain a pH-shifted soybean protein isolate solution; S3, under the condition of ice water bath, ultrasonic treatment is carried out on the pH-shifted soybean protein isolate solution obtained in S2, and then centrifugal treatment is carried out under the condition of a temperature of 4 DEG C, the supernatant is taken, dialysis is carried out, and freeze-drying is carried out to obtain freeze-dried modified soybean protein isolate powder; The freeze-dried modified soybean protein isolate powder is dispersed in distilled water to obtain a modified soybean protein isolate solution; The rutin-containing ethanol solution was dropped into the modified soybean protein isolate solution under light shielding condition, and after stirring at a rotation speed of 800 rpm for 1 h, the ethanol was evaporated, and then an equal volume of distilled water was used to replace the evaporated ethanol. After centrifugation, the supernatant was reserved to obtain a rutin-modified soybean protein isolate aqueous solution; S4, pectin was dissolved in deionized water, and after ultrasonic treatment, a pectin aqueous solution with a mass fraction of 0.1% was obtained; S5, the rutin-modified soybean protein isolate aqueous solution obtained in S3 and the pectin aqueous solution with a mass fraction of 0.1% obtained in S4 were mixed and stirred to obtain a rutin-modified soybean protein isolate-pectin aqueous solution; S6, genipin was added to the rutin-modified soybean protein isolate-pectin aqueous solution obtained in S5, and after magnetic stirring, the solution was reacted under light shielding at a temperature of 37℃ for 24 h to obtain a rutin-modified soybean protein isolate-pectin-genipin aqueous solution with a genipin concentration of 0.05 mg / mL; S7, the rutin-modified soybean protein isolate-pectin-genipin aqueous solution with a genipin concentration of 0.05 mg / mL obtained in S6 was freeze-dried to obtain a rutin-modified soybean protein isolate-pectin-genipin nanoparticle carrier.

[0007] Preferably, the stirring time in S1 is 2 h.

[0008] Preferably, the mass concentration of the modified soybean protein isolate solution in S3 is 1 mg / mL. The preparation method of the rutin-containing ethanol solution is as follows: rutin is dissolved in an ethanol solution with a volume fraction of 60% to obtain the rutin-containing ethanol solution, and the mass concentration of rutin in the rutin-containing ethanol solution is 1.5 mg / mL.

[0009] Preferably, the ultrasonic treatment power in S3 is 360 W, and the time is 5 min; the MWCO of the dialysis bag used in dialysis in S3 is 3500 Da, and the dialysis time is 24 h; the freeze-drying temperature in S3 is -80℃, and the freeze-drying time is 48 h.

[0010] Preferably, the mass concentration of rutin in the rutin-modified soybean protein isolate aqueous solution in S3 is 1.5 mg / mL, and the mass concentration of modified soybean protein isolate in the rutin-modified soybean protein isolate aqueous solution is 1 mg / mL.

[0011] Preferably, the ultrasonic treatment power in S4 is 600 W, and the time is 5 min.

[0012] Preferably, the mass ratio of the rutin-modified soybean protein isolate aqueous solution to the pectin aqueous solution with a mass fraction of 0.1% in S5 is 1:1.

[0013] Preferably, the temperature of freeze-drying in S7 is -80℃, and the time of freeze-drying is 48h; the average particle size of the rutin-modified soybean protein isolate-pectin-gossypol nanoparticle carrier in S7 is 241.6 nm.

[0014] Preferably, the encapsulation efficiency of the rutin-modified soybean protein isolate-pectin-gossypol nanoparticle carrier in S7 is 94.63%, the bioavailability is 87.86%, the rutin retention rate at 4h and 100℃ is 73.46%, the rutin retention rate after 21 days of room temperature storage is 70.82%, and the rutin-modified soybean protein isolate-pectin-gossypol nanoparticle carrier has UV stability.

[0015] The application also provides a use of the rutin-modified soybean protein isolate-pectin-gossypol nanoparticle carrier prepared by the preparation method, and the rutin-modified soybean protein isolate-pectin-gossypol nanoparticle carrier is used for gastric drug sustained release and small intestinal drug targeted release.

[0016] The pectin (PC) in the application belongs to polysaccharide substances, and can also be chitosan (CS) or guar gum; Compared with the prior art, the application has the following advantages: In the application, the modified soybean protein isolate can interact with rutin to form a non-covalent good water-soluble delivery carrier, and effectively improve the bioavailability of rutin; on this basis, by compounding a negative charged polysaccharide pectin, a rutin-modified soybean protein isolate-polysaccharide nanoparticle carrier with a "shell-core" structure can be formed; after ultrasonic treatment, the soybean protein is compounded with rutin, rutin is located in the core of the structure, and then the rutin is compounded with pectin, pectin is located in the outer shell of the particle carrier; finally, gossypol is compounded, and gossypol is located in the outermost layer of the particle carrier, which effectively resists gastric acid erosion and is targeted to be delivered to the small intestine.

[0017] The application will be further described in detail in combination with the drawings and examples. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 Fig. 1 is a graph of particle size (a), potential (b) and dispersion index (c) of the nanoparticle carrier of Example 1 and Comparative Examples 1-4 of the application.

[0019] Note: Figure 1 In the a and b graphs, the lowercase letters represent significant differences at the p<0.05 level.

[0020] Figure 2 Fig. 2 is a transmission electron microscope graph of the rutin-modified soybean protein isolate-pectin-gossypol nanoparticle carrier prepared in Example 1 of the application (the right graph is a partial enlarged view of the left graph).

[0021] Figure 3 Figure 1 is the encapsulation efficiency of rutin by the nanoparticle carrier of Example 1 and Comparative Examples 1-4 of the present application. Note: the lower case letters in the figure indicate the significant difference at the level of p<0.05.

[0022] Figure 4 Figure 2 is the rutin retention rate after thermal stability treatment (a), UV stability treatment (b) and storage stability treatment (c) of the nanoparticle carrier of Example 1 and Comparative Examples 1-4 of the present application. Note: Figure 4 The lower case letters in the figure indicate the significant difference at the level of p<0.05.

[0023] Figure 5 Figure 3 is the bioavailability of the nanoparticle carrier of Example 1 and Comparative Examples 1-4 of the present application. Note: the lower case letters in the figure indicate the significant difference at the level of p<0.05.

[0024] Figure 6 Figure 4 is the mucosal interaction (binding rate and penetration rate) (a) and mucosal penetration efficiency of rutin (b) of the nanoparticle carrier of Example 1 and Comparative Examples 1-4 of the present application. Note: the lower case letters in the figure indicate the significant difference at the level of p<0.05.

[0025] Figure 7 Figure 5 is the change of body weight of mice in each group (a) and the release of rutin in the stomach, intestine, etc. of mice (b) of the 0.05 CSR prepared in Example 1 at 1-3h after gavage of physiological saline (blank group), 300μL of modified soybean protein isolate solution (control group 1) without rutin at a concentration of 1mg / mL, 300μL of natural rutin solution (control group 2) at a concentration of 1.5mg / mL and 3 groups of Example 1 (groups 1-3 were gavaged with 100, 300 and 500μL, respectively). DETAILED DESCRIPTION

[0026] Example 1 The preparation method of the rutin-modified soybean protein isolate-pectin-gossypetin nanoparticle carrier of the present example is as follows: S1, dispersing soybean protein isolate powder in distilled water, stirring for 2h and keeping the pH value of the system at 7.0 to obtain a soybean protein isolate solution with a mass concentration of 10mg / mL; The soybean protein isolate powder in the present example was commercially available and purchased from Hefei Bomei Biological Company; S2, adjusting the pH value of the soybean protein isolate solution with a mass concentration of 10mg / mL obtained in S1 to 12.0 with sodium hydroxide aqueous solution, stirring for 1h, then adjusting the pH value to 7.0, stirring for 20min to obtain the pH-shifted soybean protein isolate solution; S3, the pH shifted soybean protein isolate solution obtained in S2 was subjected to ultrasonic treatment under the condition of ice water bath and power of 360 W for 5 min, and then centrifuged (10000 x g, 15 min) under the condition of temperature of 4 ℃, the supernatant was taken, dialyzed in a dialysis bag with MWCO of 3500 Da for 24 h, and then freeze-dried under the condition of temperature of -80 ℃ for 48 h to obtain modified soybean protein isolate powder after freeze-drying (named as SPI); The modified soybean protein isolate powder after freeze-drying was dispersed in distilled water to obtain a modified soybean protein isolate solution with a mass concentration of 1 mg / mL (named as SPI solution); Rutin was dissolved in a 60% ethanol solution by volume fraction to obtain a rutin-containing ethanol solution with a rutin mass concentration of 1.5 mg / mL; under the condition of light shielding, the rutin-containing ethanol solution was dropped into the modified soybean protein isolate solution by a needle tube, stirred at a rotation speed of 800 rpm for 1 h, and then evaporated in a rotary evaporator to evaporate ethanol, and an equal volume of distilled water was used to replace the evaporated ethanol, and the lower layer of unencapsulated rutin precipitate was removed by centrifugation at 2500 x g for 15 min, and the supernatant was retained to obtain a rutin-modified soybean protein isolate-containing aqueous solution; the rutin mass concentration in the rutin-modified soybean protein isolate-containing aqueous solution was 1.5 mg / mL, and the modified soybean protein isolate mass concentration in the rutin-modified soybean protein isolate-containing aqueous solution was 1 mg / mL; S4, pectin was dissolved in deionized water, and ultrasonic treatment was performed under the condition of power of 600 W for 5 min to obtain a pectin aqueous solution with a mass fraction of 0.1%; S5, the rutin-modified soybean protein isolate-containing aqueous solution obtained in S3 and the pectin aqueous solution with a mass fraction of 0.1% obtained in S4 were mixed and stirred at a mass ratio of 1:1 to obtain a rutin-modified soybean protein isolate-pectin-containing aqueous solution; S6, genipin was added to the rutin-modified soybean protein isolate-pectin-containing aqueous solution obtained in S5, and magnetic stirring was performed, and then the rutin-modified soybean protein isolate-pectin-containing aqueous solution was reacted under the condition of temperature of 37 ℃ and light shielding for 24 h to obtain a rutin-modified soybean protein isolate-pectin-genipin-containing aqueous solution with a genipin concentration of 0.05 mg / mL; S7, the rutin-modified soybean protein isolate-pectin-genipin-containing aqueous solution with a genipin concentration of 0.05 mg / mL obtained in S6 was freeze-dried under the condition of temperature of -80 ℃ for 48 h to obtain rutin-modified soybean protein isolate-pectin-genipin nanoparticle carriers with an average particle size of 241.6 nm, which were recorded as 0.05CSPR.

[0027] This embodiment also provides the application of a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier prepared by the above preparation method, wherein the rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier is used for sustained drug release in the stomach and targeted drug release in the small intestine.

[0028] Comparative Example 1 This comparative example describes the preparation method of rutin-modified soy protein isolate-pectin nanoparticle carrier without genipin. The method is the same as steps S1-S5 of Example 1. The aqueous solution containing rutin-modified soy protein isolate-pectin obtained in step S5 is freeze-dried at -80℃ for 48h to obtain rutin-modified soy protein isolate-pectin nanoparticle carrier without genipin, denoted as CSPR.

[0029] Comparative Example 2 The preparation method of the rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier in this comparative example is the same as that in Example 1, except that: in step S6, an aqueous solution containing rutin-modified soy protein isolate-pectin-genipin with a genipin concentration of 0.025 mg / mL is obtained, and the final rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier is denoted as 0.025CSPR.

[0030] Comparative Example 3 The preparation method of the rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier in this comparative example is the same as that in Example 1, except that in step S6, a solution containing rutin-modified soy protein isolate-pectin-genipin nanoparticles with a genipin concentration of 0.1 mg / mL is obtained. The final rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier is denoted as 0.1 CSPR.

[0031] Comparative Example 4 The preparation method of the rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier in this comparative example is the same as that in Example 1, except that in step S6, a solution containing rutin-modified soy protein isolate-pectin-genipin nanoparticles with a genipin concentration of 0.2 mg / mL is obtained. The final rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier is denoted as 0.2 CSPR.

[0032] The performance of the nanoparticle supports prepared in Example 1 and Comparative Examples 1-4 was tested: (a) Particle size, potential and dispersion index The particle size distribution was determined using a laser particle size analyzer for dynamic light scattering measurement in this example. The average diameter and polydispersity index (PDI) were calculated through light scattering measurements. The measurement results are as follows: Figure 1As shown: Particle size (a), potential (b), and dispersion index (c) of the nanoparticle carriers prepared in Example 1 and Comparative Examples 1-4. Figure 1 As shown in (a), with the increase of genipin concentration, the particle size of the nanoparticles first increases and then decreases, reaching a relatively minimum particle size in Example 1 (0.05 CSPR). This is because the cross-linking effect of genipin can stabilize pectin and soy protein isolate to a certain extent, making the structure of the nanoparticles more compact. Excessive cross-linking will lead to the aggregation of nanoparticles, thereby increasing the particle size. At the same time, the increase of genipin concentration is also accompanied by the increase of PDI, such as... Figure 1 (c); The PDI value is a dimensionless constant (range 0-1) reflecting the width of the particle size distribution. Generally, a PDI < 0.5 indicates a more uniform distribution, while a PDI > 0.5 indicates a more uneven particle size distribution and a more concentrated distribution. It is noteworthy that there is no significant difference in PDI values ​​between CSPR and 0.025CSPR and 0.05CSPR (PDI < 0.4), indicating that the crosslinking effect of low concentrations of genipin does not affect the dispersibility of nanoparticles. However, the PDI increases significantly at 0.1CSPR and 0.2CSPR. Figure 1 (c); The particle size distribution also shows a multi-peak pattern, such as... Figure 1 (a) Potential can reflect the electronegativity and stability of nanoparticles. All nanoparticle-treated groups have negatively charged surfaces, such as... Figure 1 As shown in (b), the 0.2CSPR group was significantly smaller than the other groups, indicating that excessive cross-linking leads to aggregation and thus increases the surface charge content. In summary, the measurement results showed that the 0.05CSPR particle size in Example 1 was 241.6 nm, reaching the nanoscale, and the nanoparticles had a negative surface charge, which was the same as the charge of the gastric mucosa, thus achieving electrostatic repulsion and effectively preventing adsorption and erosion of the gastric mucosa.

[0033] (II) Observation of the structure of the rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier prepared in Example 1 using transmission electron microscopy (TEM). Figure 2 (Left image), and further magnified ( Figure 2 As shown in the right figure, the outer layer of the composite nanoparticles forms a dense shell, and the interior is uniformly encapsulated with rutin, which once again proves the formation of the "shell-core" structure.

[0034] (III) The encapsulation efficiency of genipin for rutin in the nanoparticle carriers of Example 1 and Comparative Examples 1-4 was determined to evaluate the rutin encapsulation effect.

[0035] Take 1 mL of an aqueous solution containing rutin-modified soy protein isolate-pectin-genipin (0.05 mg / mL genipin concentration obtained in step S6 of Example 1), add 1 mL of 60% ethanol solution, centrifuge at 5000×g for 15 min, and use a plastic dropper to collect the extracted rutin from the upper layer. Repeat the operation twice to extract all the rutin. Treat the obtained rutin using the sodium nitrite-aluminum nitrate colorimetric method, and measure the absorbance at 500 nm using a UV-8000 spectrophotometer.

[0036] By establishing the standard curve (y = 0.1367x + 0.0095, R0), 2 = 0.994) The content of encapsulated rutin in modified soy protein isolate was determined, and the encapsulation efficiency was calculated by the following formula.

[0037] Encapsulation efficiency (%) = Encapsulated rutin (mg) / Total rutin added (mg) × 100 like Figure 3 As shown, the encapsulation efficiency first increased and then decreased with the increase of genipin. The encapsulation effect of 0.025 CSPR (86.87%) in Comparative Example 2 was better than that of CSPR in Comparative Example 1, indicating that adding genipin can effectively protect nanoparticles. Among them, the encapsulation effect of 0.05 CSPR in Example 1 was the best, with an encapsulation efficiency of 94.63%, which was 7.81% higher than that of Comparative Example 1. The encapsulation efficiency of Comparative Example 3 was 85.03%, and that of Comparative Example 4 was 82.51%. This is because with the increase of genipin addition, the degree of cross-linking intensifies, and soybean protein aggregates to varying degrees. A small amount of rutin encapsulated in shallow hydrophobic residues may escape, resulting in a decrease in encapsulation efficiency.

[0038] According to the gastrointestinal release theory of bioactive substances, the bioavailability of encapsulated rutin is determined by the rutin content (F) in the encapsulation carrier. C ), gastrointestinal release (F) B ), mucosal penetration (F) A ) and the effective dose for maintaining the bioactivity of rutin after metabolism (F M The efficiency of rutin encapsulation in the packaging carrier is therefore examined, as it is jointly determined by the factors involved. Figure 3 The measurement results showed that the encapsulation efficiency of Example 1 was significantly greater than that of Comparative Examples 1-4, combined with the results of bioaccessibility ( Figure 5 This indicates that the embodiments are beneficial for the release and application of rutin.

[0039] Therefore, the encapsulation efficiency of the rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier prepared in Example 1 reached 94.63%.

[0040] (iv) Thermal stability, UV stability and storage stability tests of the nanoparticle carriers of Examples 1 and Comparative Examples 1-4.

[0041] (1) Determine the thermal stability of the nanoparticle carriers of Example 1 and Comparative Examples 1-4.

[0042] The nanoparticles were heated at 100°C for 4 hours, and the residual amount of rutin in the nanoparticles was measured after 0, 1, 2, 3, and 4 hours of heating. The thermal stability was assessed by measuring the rutin retention rate of the nanoparticle carriers in Examples 1 and Comparative Examples 1-4 after heating for 4 hours, to ensure their suitability for food thermal processing.

[0043] The rutin retention rates of the nanoparticle carriers in Examples 1 and Comparative Examples 1-4 after heat treatment for 4 hours are as follows: Figure 4 As shown in (a), compared with the CSPR of Comparative Example 1, the thermal stability of the 0.05 CSPR of Example 1 was significantly increased. After being heated at 100°C for 4 h, the rutin retention rate of the 0.05 CSPR of Example 1 was 73.46%. Furthermore, as... Figure 4 (a) In the early heating phase (0-1 h), the rutin leakage rate of Examples 1 and Comparative Examples 2-4 with added genipin was slower than that of Comparative Example 1 without added genipin. This is because the covalent bonds formed after cross-linking genipin restrict the movement of molecular chains, which is beneficial to the compactness and stability of the tertiary network structure of nanoparticles. However, when the concentration of genipin is too high, as in Comparative Examples 3-4, there is a problem of excessive cross-linking and nanoparticle aggregation. Therefore, in the later heating phase, the rutin leakage rate of Comparative Examples 3-4 even exceeds that of Comparative Example 1. Figure 4 (a). Figure 4 (a) The data show that Example 1 can effectively improve the thermal stability of the rutin-loaded nanoparticles by using low-concentration cross-linked genipin, effectively avoiding the problem of rutin leakage caused by temperature changes in food heat processing, daily transportation and storage, so as to improve the wide range of applications.

[0044] (2) The UV stability of the nanoparticle carriers of Example 1 and Comparative Examples 1-4 was determined.

[0045] Rutin residues were measured under two UV lamps after 0, 1, 2, 3, and 4 hours of light exposure at room temperature. Figure 4As shown in (b), Examples 1 and Comparative Examples 2-4, with added genipin, showed significantly better protection against UV irradiation of rutin than Comparative Example 1. This is mainly attributed to three factors: besides the efficient UV absorption capacity of the aromatic amino acids in soy protein isolate and the light scattering and absorption mechanism of the cross-linked composite shell, the blue-purple complex generated after cross-linking genipin, which deepens the color of the sample, also effectively protects the aromatic ring structure of the encapsulated rutin from UV decomposition. Among them, the nanoparticle carrier of Example 1 exhibited the best UV protection performance, such as... Figure 4 (b). Therefore, the bioactivity of rutin can be effectively protected during application delivery.

[0046] (3) Determine the storage stability of the nanoparticle carriers of Example 1 and Comparative Examples 1-4.

[0047] The retention rate of rutin in the samples was measured at room temperature for 0, 1, 3, 5, 7, 14, and 21 days to evaluate the storage stability of the examples. Figure 4 As shown in (c), compared to Comparative Example 1, Example 1 exhibited stronger storage stability, with almost no loss during storage from 14 to 21 days. After 21 days of storage, the nanoparticle carrier of Example 1 still retained 70.82% of rutin. Comparative Example 2 showed no significant difference in rutin content at the end of storage compared to Example 1, but the decreasing trend in rutin content was more pronounced during the short-term storage period (the first 7 days). This may be due to the lower genipin concentration and lower degree of cross-linking in Comparative Example 2 compared to Example 1. Comparative Examples 1 and 3-4, however, showed a more significant decreasing trend throughout the entire storage period. Figure 4 (c). Therefore, by optimizing storage stability using the method of Example 1, the short-term and long-term shelf life of the product can be effectively extended and the product supply can be improved.

[0048] (v) Determination of the bioaccessibility of nanoparticle carriers in Example 1 and Comparative Examples 1-4.

[0049] The bioaccessibility of the nanoparticle carriers of Example 1 and Comparative Examples 1-4 was determined using an in vitro semi-static gastrointestinal digestion model, reflecting the potential ability of the nanoparticle carriers prepared in Example 1 to release rutin during gastrointestinal digestion.

[0050] Simulated gastric juice (SGF, pH 1.11) was prepared using pepsin (0.32%, w / v), hydrochloric acid (0.085 mol / L), and NaCl (0.03 mol / L).

[0051] The original formulation of simulated intestinal fluid (SIF) included the use of 0.05 mol / L KH2PO4, 10 mg / mL of trypsin, and pH adjustment to 6.8 using 0.2 mol / L NaOH aqueous solution.

[0052] The nanoparticle carriers of Example 1 and Comparative Examples 1-4 were dispersed in water to obtain nanoparticle carrier sample solutions with a concentration of 1 mg / mL. 5 mL of each sample solution was added to SGF (simulated gastric juice) at a volume ratio of 1:1 and gently mixed at 37±0.5℃ for 120 min (i.e., simulated gastric digestion for 2 h) to obtain a mixed solution of equal volume (10 mL) of sample solution and simulated gastric juice, which is the aliquot sample solution (pH 7.4).

[0053] In addition, 10 mL of SIF was added to 10 mL of the above aliquot sample solution (pH 7.4), and the mixture was stirred at 37 ± 0.5 °C for 120 min. Finally, the mixture was centrifuged at 8500 rpm for 30 min at 4 °C. The supernatant was then collected and diluted 5-fold with 60% ethanol solution, and the concentration was obtained by measuring the absorbance of the diluted sample.

[0054] The following formula is used to calculate biological accessibility: Bioavailability (%) = Rutin release (mg) / Total rutin added (mg) × 10; The results are as follows Figure 5 As shown, during the 4-hour gastrointestinal digestion phase, the rutin release trends of Comparative Examples 2-4 were similar to those of Example 1. Rutin showed good protection in gastric juice, while in intestinal juice, it rapidly released rutin in response to pH changes. However, the CSPR prepared in Comparative Example 1 exhibited significantly higher rutin release efficiency than Example 1 during the first 2 hours of gastric digestion, releasing 41.39% of rutin after 2 hours of simulated gastric digestion. The release efficiency in the intestine, however, slowed down, resulting in a final bioavailability of 78.97%. Example 1, on the other hand, showed the opposite: slow release during gastric digestion, but rapid rutin release in the intestine, achieving a bioavailability of 87.86% after 4 hours of gastrointestinal digestion.

[0055] This demonstrates that the preparation method of Example 1 can effectively ensure the sustained-release effect of rutin. This is mainly due to the fact that genipin in the outer "shell" structure of the nanoparticles in Example 1 effectively maintains the stability of the overall structure, thereby effectively blocking the influence of the extreme acidic pH environment of the stomach, preventing the delivery system from becoming unbalanced, and effectively delaying the release of rutin. This proves that the 0.05 CSPR prepared in Example 1 has the characteristics of sustained-release and targeted intestinal delivery. Comparative Examples 2-4 also have similar "shell" structures, but different genipin concentrations have different effects on the stability of the cross-linked nanoparticles. Therefore, their final biotechnological significance is less than that of Example 1. Among them, the 0.2 CSPR prepared in Comparative Example 4, due to its higher degree of cross-linking, did not completely release the rutin inside at the end of digestion in the small intestine.

[0056] (vi) The diffusion behavior of the nanoparticle carriers of Example 1 and Comparative Examples 1-4 in the gastrointestinal mucosa was determined by using the Transwell chamber experiment.

[0057] The specific method is as follows: The Transwell chamber contains a donor chamber and a recipient chamber, separated by a polycarbonate semi-permeable membrane, allowing nanoparticles to pass through but preventing cell and mucosal permeation. A 12-well Transwell chamber (3 μm pore size) was selected. 0.3 g of porcine colonic mucus was added to the semi-permeable membrane. 1.5 mL of PBS (0.05 mol / L, pH 7.0) was added to the recipient chamber. 0.5 mL of the nanoparticle carrier sample solution (1 mg / mL) from Examples 1 and Comparative Examples 1-4 was added to the donor chamber. The device was incubated at 37°C and 100 rpm for 2 h on a shaker. 0.2 mL of sample solution was taken from the recipient chamber, and its fluorescence intensity (FITC, excitation = 488 nm, emission = 520 nm) was read. The penetration efficiency of the nanoparticle carriers from Examples 1 and Comparative Examples 1-4 in the intestinal mucosa was calculated using the following formula: Mucosal penetration rate (%) = (Intensity) supebasolateral -Intensity blank Intensity sample ×100; Among them, Intensity supebasolateral Intensity represents the fluorescence intensity of the sample solution in the acceptor chamber. blank The fluorescence intensity of the blank control is shown in the figure. sample The fluorescence intensity represents the fluorescence intensity of the nanoparticle carrier sample solution delivery system obtained by dispersing the nanoparticle carriers prepared in Example 1 and Comparative Examples 1-4 in water.

[0058] To more intuitively characterize the mucosal transport effect of rutin, the solution in the above receptor chamber was centrifuged at 5000×g for 15min, and the rutin content was determined according to the encapsulation efficiency method.

[0059] (1) The penetration effect of the mucosa is also one of the important factors affecting the bioavailability of drug delivery. Therefore, the penetration amount (F) of the mucosa in Example 1 and Comparative Examples 1-4 was evaluated. A The interaction between nanoparticles and the intestinal mucosa was investigated, and characterized by penetration efficiency and binding efficiency. The results are as follows: Figure 6 As shown in (a), with the increase of genipin concentration (genipin concentration: Comparative Example 1 < Comparative Example 2 < Example 1 < Comparative Example 3 < Comparative Example 4), the mucosal penetration rate of the nanoparticle carrier gradually decreased, while the mucosal binding rate increased. It is worth noting that there was no significant difference between Comparative Examples 1-2 and Example 1. This is because the low concentration of genipin crosslinking reaction does not affect the mucosal penetration effect of nanoparticles; however, the high concentration of genipin crosslinking reaction has a greater impact. The 0.2CSPR prepared in Comparative Example 4 had the lowest mucosal penetration rate (22.55%) and the highest mucosal binding rate (38.70%), indicating that excessive crosslinking promotes the aggregation of nanoparticles, resulting in nanoparticles with excessively large particle sizes that can only adhere to and remain on the outer layer of mucus, thus reducing the mucosal penetration ability.

[0060] (2) In order to further investigate the mucosal penetration effect of nanoparticles, the permeability of rutin after nanoparticles pass through the membrane was measured. The sample solution in the receptor chamber after incubation for 2 hours after the mucosal penetration experiment in (VI) was centrifuged at 5000 ×g for 15 min, and the rutin content was determined according to the method for determining rutin content in (III).

[0061] The results are as follows Figure 6 As shown in (b), the trend of rutin permeability in nanoparticles prepared in each group in the receptor chamber and Figure 6 (a) The trends of the permeability of each nanoparticle are basically consistent, showing a significant decrease with increasing genipin concentration. However, the permeability of rutin in Example 1 is significantly higher than that in Comparative Examples 1-2. The rutin permeability of the 0.05 CSPR prepared in Example 1 is higher, at 47.59%, which is significantly higher than that of the CSPR in Comparative Example 1. The rutin permeability of the nanoparticle carrier in Comparative Example 2 is relatively lower, at 44.43%, which is not significantly different from that in Comparative Example 1. This result is somewhat different from the permeability of the nanoparticles, which may be due to the relatively high rutin content encapsulated in the 0.05 CSPR prepared in Example 1.

[0062] The results in summary indicate that the nanoparticle carrier 0.05 CSPR prepared in Example 1 is beneficial for improving the permeability of the intestinal mucosa of the delivery system.

[0063] (vii) Release kinetics simulation of the release in the gastrointestinal tract in Example 1. Samples were collected and analyzed periodically during in vitro gastrointestinal digestion. In short, samples were collected at 15, 30, 60, 90, 120, 150, 180, 210, and 240 min. The cumulative release rate of rutin was measured according to the following formula: Cumulative release (%) = Total rutin released / Total rutin added × 100 To investigate the release mechanism of rutin encapsulated in nanoparticles, release data were fitted to four different kinetic models: Zeroth-order equation: Q t / Q ∞ =k0t Where Qt represents the cumulative release of rutin at time t, and Q... ∞ This indicates that the encapsulated rutin is cumulatively released from the SPI nanoparticles until complete release is achieved. k0 represents the zero-order release constant.

[0064] First-order equation: -ln(1-Q) t / Q ∞ =k1t Qt / Q ∞ It represents the proportion of rutin released, and k1 represents the first-order release constant.

[0065] Higuchi equation: Q t / Q ∞= k H t 1 / 2 Qt / Q ∞ The proportion of rutin released, k H This represents the dissolution constant in the Higuchi equation.

[0066] Korsmeyer-Peppas equation: Q t / Q ∞= kt n Where n is the diffusion exponential property of the Korsmeyer-Peppas mode, k is the release constant, and Qt / Q ∞ It refers to the proportion of rutin released.

[0067] In the field of bioactive compound delivery, release kinetics analysis is widely used to determine drug delivery patterns by fitting various mathematical models. To investigate the release kinetics of rutin in 0.05 CSPR prepared in Example 1, in vitro release data were obtained and analyzed using four different kinetic models (zero-order, first-order, Higuchi, and Korsmeyer-Peppas models). Table 1 shows that, except for the Higuchi model in the intestine, the R0.05 of 0.05 CSPR varies in each model. 2 All values ​​were greater than 0.9, indicating a good fit. Among them, 0.05 CSPR showed an excellent fit (R²) to the first-order equation in gastric juice. 2 >0.99), rutin release follows a typical time-dependent relationship, meaning the amount of rutin released changes exponentially with time, consistent with the characteristics of sustained-release formulations of drugs with continuous slow release; and in the intestine, it best fits the zero-order equation (R0). 2 The n-value (>0.99) indicates that the release of rutin in the intestine at 0.05 CSPR follows a concentration gradient release pattern, consistent with the characteristics of a sustained-release formulation that gradually releases bioactive compounds at a constant rate. The release mechanism was further explored using the Korsmeyer-Peppas model. The n-value, a release index, is crucial in describing the release mechanism, and the release indices are shown in Table 2. According to the results in Table 1, the n-values ​​in the stomach and intestine of Example 1 were 0.64 and 0.8931, respectively. Corresponding to the release mechanism in Table 2, this indicates that the encapsulated rutin undergoes non-free diffusion in the stomach, while in the intestine, free diffusion and skeletal dissolution coexist. The n-value in the intestine, 0.8931, is closer to 0.89, suggesting that skeletal dissolution is the dominant mechanism in this nanoparticle. Therefore, a comprehensive evaluation of the SPI composite carrier during gastrointestinal digestion shows that swelling digestion is the dominant process, accompanied by a small amount of rutin diffusion and release, demonstrating the sustained-release effect of this carrier system in the stomach. In contrast, the carrier prepared in Example 1 undergoes complete skeletal dissolution in the intestine, meaning that the nanocarrier shell rapidly decomposes upon entering the intestine, releasing a large amount of rutin, thus increasing bioavailability. In summary, the rutin in Example 1 is slowly released in gastric juice and rapidly released at a relatively constant rate in intestinal juice. These studies indicate that Example 1 provides good protection and sustained-release of rutin during digestion, which is beneficial for targeted release into the intestine.

[0068] Table 1. Release kinetic parameters of rutin in 0.05 CSPR prepared in Example 1 in the stomach and intestine. Note: The "-" symbol indicates that the equation does not have this parameter.

[0069] Table 2. Release mechanism of rutin at 0.05 CSPR prepared in Example 1 in the stomach and intestine. (VIII) Thirty-six clean-grade, 6-week-old male ICR mice (weighing 20g ± 1g) were housed in an SFP-grade animal facility. The animal experiments were approved by the Animal Welfare and Ethics Committee of Harbin University of Commerce and conducted at the Animal Experiment Center (SYXK (Heilongjiang) 2023-002). All experimental procedures adhered to internationally and nationally accepted animal welfare and experimental principles. The housing temperature was controlled at 23 ± 1℃, humidity at 75% ± 5%, with a 12-hour light / 12-hour dark cycle. Mice had free access to water and food. After one week of acclimatization feeding with standard feed, gavage administration began. Specific details are shown in Table 3.

[0070] Table 3. Comparison of mouse grouping and daily treatment Note: Control group 1 modified soy protein isolate solution is obtained by dispersing 1 mg of freeze-dried modified soy protein isolate powder obtained through S1-S3 in 1 mL of water; Control group 2 natural rutin solution is obtained by dissolving 1.5 mg of natural rutin in 1 mL of distilled water. The solution in Example 1 is the 0.05 CSPR solution prepared in step S6 of Example 1.

[0071] Mice were sacrificed 1, 2, and 3 hours after the final gavage. The stomach, stomach contents, intestine, and intestinal contents were collected, and a measured amount of physiological saline (1:2, w / v) was added. The mixture was homogenized into a homogeneous tissue fluid. 200 μL of the tissue homogenate was accurately measured and placed in a 1.5 mL centrifuge tube. 100 μL of tissue extract was added, and the mixture was vortexed for 1 min, then centrifuged at 12000 rpm for 10 min.

[0072] The chromatographic column was a ZORBAX SB-C. 18 (250 mm × 4.6 mm, 5 μm), 1.0 mL / min; column temperature 40 ℃; detection wavelength 360 nm; mobile phase: methanol (A): 0.4% phosphoric acid water = 70:30, elution gradient (0~5 min, 50%~50%; 5~10 min, 30%~70%; 10~11 min, 10%~90%; 11~12 min, 70%~30%); injection volume 10 μL.

[0073] The standard curves for rutin samples of different concentrations are plotted as follows: y = 29.87x + 1.50, R0 2 =0.9993.

[0074] (1) Figure 7(a) Changes in body weight of mice after gavage administration of 300 μL of physiological saline (blank group), 300 μL of modified soy protein isolate solution without rutin (control group 1), 300 μL of natural rutin solution (control group 2), and different doses of 0.05 CSPR (100 μL, 300 μL, 500 μL) of Example 1.

[0075] The modified soy protein isolate solution without rutin in control group 1 was prepared as follows: 1 mg of freeze-dried modified soy protein isolate powder obtained in steps S1-S3 of Example 1 was dispersed in 1 mL of distilled water to obtain a modified soy protein isolate solution of 1 mg / mL, which was designated as control group 1.

[0076] The preparation method of the natural rutin solution in control group 2 is as follows: 1.5 mg of natural rutin solid powder is dispersed in 1 mL of distilled water to obtain a natural rutin solution of 1.5 mg / mL, which is referred to as control group 2.

[0077] All mice showed an increasing weight trend, indicating that they were in their growth period and that the 0.05 CSPR prepared in Example 1 had a good taste. No nausea or anorexia was observed in the mice after gavage, suggesting that Example 1 has good prospects for application in the food industry. It is noteworthy that the modified soy protein solution without rutin prepared in Control Group 1, whose main component is modified soy protein, had a significantly higher weight than other groups throughout the feeding period, indicating that rutin has a potential role in controlling weight. Furthermore, the weight of mice in Example 1, which received 300 μL of the solution via gavage, was significantly lower than other groups throughout the feeding period. These results demonstrate that the rutin in Example 1 was effectively released in the intestines, and that at an appropriate gavage dose, Example 1 has the potential to control mouse weight.

[0078] (2) To further investigate whether the rutin in Example 1 was released in mice as planned, 300 μL of the solution from Example 1 was administered by gavage, and the rutin content in the stomach, stomach cavity, intestine, and intestinal cavity of mice was measured at 1, 2, and 3 hours. The results are as follows: Figure 7 As shown in (b), experimental data revealed significant changes in the distribution dynamics of rutin after drug administration: at 1 hour, rutin was mainly concentrated in the stomach, while after 3 hours it migrated to the intestinal region, at which point the highest concentration (15.89 μg / mL) was detected in the intestinal contents. The time to reach this peak concentration is highly consistent with the characteristics of in vitro cumulative release studies, demonstrating that the delivery system has the efficacy of delaying drug release, and its regulated release pattern is in line with the design expectations of an intestinal target delivery system.

[0079] Example 1 prepared a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier for sustained drug release in the stomach and targeted drug release in the small intestine. This nanocarrier has advantages such as small particle size, high encapsulation efficiency, good stability and high mucosal permeability.

[0080] In summary, the soy protein isolate in this invention can interact with rutin to form a non-covalent, well-water-soluble delivery carrier, effectively improving the bioavailability of rutin. Based on this, by compounding with neutral or negatively charged polysaccharides and then covalently crosslinking with genipin, a "shell-core" structure of rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier can be formed, which effectively resists gastric acid erosion and is targeted to the small intestine.

[0081] The delivery carrier matrix in this invention comprises naturally derived food-grade raw materials including soy protein, pectin, and genipin. Rutin, the target of the encapsulation, is a hydrophobic flavonoid found in fruits and vegetables, possessing clear health benefits in anti-inflammatory, sedative, and anti-tumor effects. However, due to its extremely low permeability and stability, rutin's direct oral absorption is poor, limiting its application in the food industry. The rutin-soy protein-pectin-genipin nanoparticles of this invention effectively address these issues. Encapsulation forms a stable "shell-core" structure, improving thermal stability (73.46% rutin retention after 4 hours), storage stability (70.82% rutin retention after 21 days), and UV stability. Appropriate cross-linking tightens the particle structure, achieving a mucosal penetration rate of 47.69%. The nanoparticles release rutin primarily through gastric skeleton dissolution with diffusion as a secondary process, while in the intestine, complete skeleton dissolution occurs. Therefore, rutin can be released in large quantities in the intestine, achieving targeted intestinal delivery. In vitro and in vivo digestion studies show consistent results, significantly enhancing the intestinal delivery efficiency of rutin.

[0082] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any way. Any simple modifications, alterations, and equivalent changes made to the above embodiments based on the inventive essence shall still fall within the protection scope of the present invention.

Claims

1. A method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier, characterized in that, The method is as follows: S1. Disperse soy protein isolate powder in distilled water, stir and maintain the pH of the system at 7.0 to obtain a soy protein isolate solution with a mass concentration of 10 mg / mL; S2. Adjust the pH of the 10 mg / mL soy protein isolate solution obtained in S1 to 12.0 with sodium hydroxide aqueous solution, stir for 1 h, then adjust the pH to 7.0 and stir for 20 min to obtain the soy protein isolate solution with pH shift. S3. The soy protein isolate solution with pH shift obtained in S2 is subjected to ultrasonic treatment under ice-water bath conditions, centrifuged at 4°C, the supernatant is collected, dialyzed, and then freeze-dried to obtain freeze-dried modified soy protein isolate powder. The freeze-dried modified soy protein isolate powder was dispersed in distilled water to obtain a modified soy protein isolate solution; Under light-protected conditions, an ethanol solution containing rutin was added dropwise to the modified soy protein isolate solution. After stirring at 800 rpm for 1 h, the ethanol was evaporated, and an equal volume of distilled water was used to replace the evaporated ethanol. After centrifugation, the supernatant was retained to obtain an aqueous solution containing rutin-modified soy protein isolate. S4. Dissolve pectin in deionized water, and after ultrasonic treatment, obtain a pectin aqueous solution with a mass fraction of 0.1%. S5. Mix and stir the aqueous solution containing rutin-modified soy protein isolate obtained in S3 and the aqueous solution with a mass fraction of 0.1% pectin obtained in S4 to obtain an aqueous solution containing rutin-modified soy protein isolate-pectin. S6. Add genipin to the aqueous solution containing rutin-modified soy protein isolate-pectin obtained in S5, stir magnetically, and react in the dark at 37℃ for 24 hours to obtain an aqueous solution containing rutin-modified soy protein isolate-pectin-genipin with a genipin concentration of 0.05 mg / mL. S7. After freeze-drying the aqueous solution containing rutin-modified soy protein isolate-pectin-genipin with a genipin concentration of 0.05 mg / mL obtained in S6, rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier is obtained.

2. The method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier according to claim 1, characterized in that, The stirring time in S1 is 2 hours.

3. The method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier according to claim 1, characterized in that, The mass concentration of the modified soy protein isolate solution described in S3 is 1 mg / mL; The ethanol solution containing rutin is prepared by dissolving rutin in a 60% ethanol solution by volume, wherein the mass concentration of rutin in the ethanol solution containing rutin is 1.5 mg / mL.

4. The method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier according to claim 1, characterized in that, The ultrasonic treatment described in S3 has a power of 360W and a duration of 5 minutes; the MWCO of the dialysis bag used in S3 is 3500 Da, and the dialysis time is 24 hours; the freeze-drying temperature in S3 is -80℃, and the freeze-drying time is 48 hours.

5. The method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier according to claim 1, characterized in that, The rutin-modified soy protein isolate aqueous solution in S3 has a rutin concentration of 1.5 mg / mL and a modified soy protein isolate concentration of 1 mg / mL.

6. The method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier according to claim 1, characterized in that, The ultrasonic treatment in S4 has a power of 600W and a duration of 5 minutes.

7. The method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier according to claim 1, characterized in that, The mass ratio of the aqueous solution containing rutin-modified soy protein isolate and the aqueous solution containing 0.1% pectin in S5 is 1:

1.

8. The method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier according to claim 1, characterized in that, The freeze-drying temperature in S7 is -80℃, and the freeze-drying time is 48h; the average particle size of the rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier in S7 is 241.6 nm.

9. The method for preparing a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier according to claim 1, characterized in that, The encapsulation efficiency of the rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier described in S7 is 94.63%, the bioavailability is 87.86%, the rutin retention rate at 4 h and 100 °C is 73.46%, and the rutin retention rate at 21 days of room temperature storage is 70.82%. The rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier has UV stability.

10. An application of a rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier prepared by the preparation method according to any one of claims 1-9, characterized in that, The rutin-modified soy protein isolate-pectin-genipin nanoparticle carrier is used for sustained drug release in the stomach and targeted drug release in the small intestine.

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