Preparation method of cholate modified beta-carotene nano solution

Abstract

The invention discloses a preparation method of a cholate modified beta-carotene nano solution, which comprises the following steps: dissolving carotene in ethyl acetate as an organic phase; dissolving protein in ultrapure water to serve as a water phase; mixing the organic phase and the water phase, shearing, dispersing, homogenizing and performing rotary evaporation to obtain a beta-carotene protein nano solution; dissolving cholate in ultrapure water, and stirring to obtain a cholate aqueous solution; and mixing and stirring a bile salt aqueous solution and the beta-carotene protein nano solution to obtain the bile salt modified beta-carotene protein nano solution. The cholate-modified beta-carotene protein nano solution provided by the invention has better gastrointestinal tract stability, can quickly penetrate through a cholate mucous layer, better crosses intestinal epithelial cells, and can effectively improve the oral bioavailability of beta-carotene; the preparation method provided by the invention is simple in operation process, safe in material, high in cost performance and convenient for large-scale production.

Description

Technical Field

[0001] The invention belongs to the technical field of food additives, and in particular relates to a method for preparing a bile salt-modified beta-carotene protein nano solution and the selection of bile salt. Background Art

[0002] β-carotene (BC) is a red, yellow or orange-red carotenoid. Carotenoids (such as β-carotene) are essential dietary components for the human body. Humans cannot synthesize them by themselves and need to be supplemented exogenously. A large number of epidemiological and clinical studies have reported that β-carotene can reduce the risk of cardiovascular disease, skin disease, cancer and ophthalmic diseases. Oral administration is the most easily accepted route of administration for patients, with the advantages of convenient administration and good safety. However, the insolubility of β-carotene in water, poor oxidative stability and low bioavailability limit its oral effect. In recent years, nanocarrier systems with natural biomacromolecules as emulsifiers have been widely studied to improve the aqueous solubility and gastrointestinal stability of fat-soluble nutrients and form controlled release in the intestine, so as to achieve the purpose of effective utilization to improve the bioavailability of nutrient molecules.

[0003] However, affected by the intestinal mucus layer and intestinal cell barrier, the cellular absorption rate of β-carotene nanocarriers will be reduced. A good oral agent should have a good ability to pass through intestinal epithelial cells and reduce the influence of enzymes, acids, alkalis and salt ions in the intestine. The structure of existing protein nano-solution carriers is destroyed after gastrointestinal digestion, resulting in nutrient leakage, increased drug loss, and increased drug toxicity to the gastrointestinal tract. Therefore, bile salts are used to modify protein nano-solution carriers, which improves the bioavailability of β-carotene to a certain extent. Summary of the invention

[0004] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0005] In view of the above problems and / or the problems existing in the prior art, the present invention is proposed.

[0006] Therefore, the purpose of the present invention is to overcome the deficiencies in the prior art and provide a method for preparing a bile salt-modified β-carotene protein nano solution.

[0007] In order to solve the above technical problems, the present invention provides the following technical solution: a method for preparing a bile salt-modified β-carotene protein nano solution, characterized in that: comprising:

[0008] The carotene was dissolved in ethyl acetate as the organic phase; the protein was dissolved in ultrapure water as the aqueous phase;

[0009] The organic phase and the aqueous phase are mixed, sheared, dispersed, homogenized, and rotary evaporated to obtain a β-carotene protein nano solution;

[0010] dissolving bile salt in ultrapure water and stirring to obtain a bile salt aqueous solution;

[0011] The bile salt aqueous solution and the beta-carotene protein nano solution are mixed and stirred to obtain the bile salt modified beta-carotene protein nano solution.

[0012] As a preferred embodiment of the preparation method of the present invention, the bile salt comprises one or more of sodium cholate, sodium deoxycholate, sodium chenodeoxycholate and naturally extracted pig bile salt.

[0013] As a preferred embodiment of the preparation method of the present invention, the concentration of the bile salt aqueous solution is 8 mg / mL.

[0014] As a preferred embodiment of the preparation method of the present invention, the addition ratio of the bile salt aqueous solution and the β-carotene protein nano solution is 1-2:2-4.

[0015] As a preferred embodiment of the preparation method of the present invention, the ratio of the added amount of β-carotene and ethyl acetate in the organic phase is 1-10:1-100.

[0016] As a preferred embodiment of the preparation method of the present invention, the protein comprises one or more of soy protein, pea protein, rice protein, peanut protein, whey protein isolate, brown rice protein and oat protein.

[0017] As a preferred embodiment of the preparation method of the present invention, the homogenization condition is homogenization once at 100 MPa in a microfluidizer.

[0018] As a preferred embodiment of the preparation method of the present invention, the volume ratio of the organic phase to the aqueous phase is 1:2-10.

[0019] Another object of the present invention is to overcome the deficiencies in the prior art and provide a method for preparing a bile salt-modified β-carotene protein nano solution to obtain a bile salt-modified β-carotene protein nano solution.

[0020] Another object of the present invention is to overcome the deficiencies in the prior art and provide a bile salt-modified β-carotene protein nano solution for use in oral β-carotene delivery.

[0021] Beneficial effects of the present invention:

[0022] The bile salt-modified β-carotene protein nano solution provided by the present invention has good biocompatibility and can efficiently encapsulate β-carotene for oral delivery; the preparation method of the bile salt-modified β-carotene protein nano solution provided by the present invention is simple and fast, and can be used for industrial production; the bile salt-modified β-carotene protein nano solution provided by the present invention has good gastrointestinal stability, can quickly penetrate the bile salt mucus layer, and can better cross the intestinal epithelial cells, and can effectively improve the oral bioavailability of β-carotene; the preparation method provided by the present invention has a simple operation process, safe materials and high cost performance, and is convenient for large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative labor. Among them:

[0024] Figure 1 The particle size diagrams of Examples 1 to 4 and Comparative Example 1 before and after digestion are shown.

[0025] Figure 2 The potential diagrams of Examples 1 to 4 and Comparative Example 1 before and after digestion are shown.

[0026] Figure 3 The cell survival rates at different dilution times after digestion of Examples 1 to 4 and Comparative Examples 1 and 2 are shown.

[0027] Figure 4 This is a graph showing the vitamin A equivalent transported into cells after digestion of Examples 1 to 4 and Comparative Examples 1 and 2.

[0028] Figure 5 The graph shows the retinol content transported into cells after digestion of Examples 1 to 4 and Comparative Examples 1 and 2.

[0029] Figure 6 The graph shows the β-carotene content transported into cells after digestion of Examples 1 to 4 and Comparative Examples 1 and 2.

[0030] Figure 7 This is a graph showing the accumulation of VA in the liver and adipose tissue of mice after digestion of Examples 1 to 4 and Comparative Examples 1 and 2. DETAILED DESCRIPTION

[0031] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the embodiments of the specification.

[0032] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0033] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0034] Unless otherwise specified, all raw materials used in the examples of the present invention are commercially available, as shown in Table 1 for details.

[0035] Table 1 Raw material name, commercial channel and product number

[0036]

[0037]

[0038] Example 1

[0039] The present invention provides a method for preparing a bile salt-modified β-carotene protein nano solution, comprising the following steps:

[0040] (1) 0.1% by mass volume fraction of β-carotene was fully dissolved in ethyl acetate as an organic phase, and 1% by mass volume fraction of soy protein was dissolved in ultrapure water as an aqueous phase. The organic phase and the aqueous phase were mixed in a volume ratio of 1:9, and dispersed at a high speed of 10000 rpm for 10 minutes, and then homogenized once by a microfluidizer under the condition of 100 MPa; the homogenized mixed suspension was passed through a rotary evaporator at 250 rpm for 1 hour to remove the organic phase in the suspension, thereby obtaining a β-carotene protein nano solution.

[0041] (2) 640 mg of sodium cholate was dissolved in 80 ml of ultrapure water and stirred at 600 rpm for two hours to obtain a bile salt solution; then the bile salt solution was mixed with the β-carotene protein nanosolution in a volume ratio of 1:2 and stirred at 600 rpm for half an hour to obtain a bile salt-modified β-carotene protein nanosolution.

[0042] Example 2

[0043] A method for preparing a bile salt-modified β-carotene protein nano solution comprises the following steps:

[0044] 0.1% mass volume fraction of β-carotene is fully dissolved in ethyl acetate as an organic phase, 1% mass volume fraction of soy protein is dissolved in ultrapure water as an aqueous phase, the ethyl acetate containing β-carotene and the aqueous phase are mixed in a volume ratio of 1:9, and after high-speed dispersion at a speed of 10000 rpm for 10 minutes, it is homogenized once by a microfluidizer under the condition of 100 MPa; the homogenized mixed suspension is passed through a rotary evaporator, and rotary evaporated at 250 rpm for 1 hour to remove the organic phase in the suspension, so as to obtain a β-carotene protein nano solution.

[0045] 640 mg of sodium deoxycholate was dissolved in 80 ml of ultrapure water and stirred at 600 rpm for two hours. Then, the bile salt aqueous solution and the β-carotene protein nano solution were mixed at a volume ratio of 1:2 and stirred at 600 rpm for half an hour to obtain a bile salt-modified β-carotene protein nano solution.

[0046] Example 3

[0047] A method for preparing a bile salt-modified β-carotene protein nano solution comprises the following steps:

[0048] 0.1% mass volume fraction of β-carotene is fully dissolved in ethyl acetate as an organic phase, 1% mass volume fraction of soy protein is dissolved in ultrapure water as an aqueous phase, the ethyl acetate containing β-carotene and the aqueous phase are mixed in a volume ratio of 1:9, and after high-speed dispersion at a speed of 10000 rpm for 10 minutes, it is homogenized once by a microfluidizer under the condition of 100 MPa; the homogenized mixed suspension is passed through a rotary evaporator, and rotary evaporated at 250 rpm for 1 hour to remove the organic phase in the suspension, so as to obtain a β-carotene protein nano solution.

[0049] 640 mg sodium chenodeoxycholate was dissolved in 80 ml ultrapure water and stirred at 600 rpm for two hours. Then the bile salt solution was mixed with the β-carotene protein nano solution at a volume ratio of 1:2 and stirred at 600 rpm for half an hour to obtain a bile salt modified β-carotene protein nano solution.

[0050] Example 4

[0051] A method for preparing a bile salt-modified β-carotene protein nano solution comprises the following steps:

[0052] 0.1% mass volume fraction of β-carotene is fully dissolved in ethyl acetate as an organic phase, 1% mass volume fraction of soy protein is dissolved in ultrapure water as an aqueous phase, the ethyl acetate containing β-carotene and the aqueous phase are mixed in a volume ratio of 1:9, and after high-speed dispersion at a speed of 10000 rpm for 10 minutes, it is homogenized once by a microfluidizer under the condition of 100 MPa; the homogenized mixed suspension is passed through a rotary evaporator, and rotary evaporated at 250 rpm for 1 hour to remove the organic phase in the suspension, so as to obtain a β-carotene protein nano solution.

[0053] 640 mg of naturally extracted pig bile salt was dissolved in 80 ml of ultrapure water and stirred at 600 rpm for two hours. Then the bile salt solution was mixed with the β-carotene protein nano solution at a volume ratio of 1:2 and stirred at 600 rpm for half an hour to obtain a bile salt-modified β-carotene protein nano solution.

[0054] Comparative Example 1

[0055] The method for preparing a bile salt-free β-carotene protein nano solution comprises the following steps:

[0056] 0.1% mass volume fraction of β-carotene is fully dissolved in ethyl acetate as an organic phase, 1% mass volume fraction of soy protein is dissolved in ultrapure water as an aqueous phase, the ethyl acetate containing β-carotene and the aqueous phase are mixed in a volume ratio of 1:9, and after high-speed dispersion at a speed of 10000 rpm for 10 minutes, it is homogenized once by a microfluidizer under the condition of 100 MPa; the homogenized mixed suspension is passed through a rotary evaporator, and rotary evaporated at 250 rpm for 1 hour to remove the organic phase in the suspension, so as to obtain a β-carotene protein nano solution.

[0057] According to Table 2, different pre-digestion β-carotene protein nano solutions have different particle sizes and potentials, wherein the order of particle sizes from large to small is Example 4> Example 1> Example 2> Example 3> Comparative Example 1. The order of potentials from large to small is Example 4> Example 3> Example 2> Example 1> Comparative Example 1.

[0058] Example 5

[0059] The method for measuring the particle size and potential of bile salt-modified β-carotene protein nano-solution comprises the following steps:

[0060] Take 50 μL of the sample to be tested and dissolve it in 5 ml of PBS standard solution, vortex and shake for 10 seconds, and then use Malvern particle size analyzer to measure its particle size and potential.

[0061] Table 2 Particle size and potential of samples of Examples 1 to 4 and Comparative Example 1 before digestion

[0062] Sample before digestion Particle size (nm) Potential (mv) Example 1 627.6±5.9 -25.12±2.3 Example 2 509.3±3.7 -27.61±1.5 Example 3 460.4±8.6 -29.96±4.6 Example 4 705.7±8.2 -33.77±3.3 Comparative Example 1 430.2±6.3 -18.35±2.7

[0063] like Figure 1 As shown, different digested β-carotene protein nano solutions have different particle sizes, which are ranked from large to small as follows: Comparative Example 1>Example 4>Example 1>Example 2>Example 3.

[0064] Example 6

[0065] The digestion of the bile salt-modified β-carotene protein nanosolution comprises the following steps:

[0066] Take 10 ml of bile salt-modified β-carotene protein nano solution sample and mix it with an equal amount of ultrapure water, add it to pepsin gastric digestion solution incubated at 37°C for 5 minutes, the amount of gastric digestion solution added is 20 ml, and the pepsin content is 26.67 mg. After digestion for 1 hour, add 40 ml of intestinal digestion solution. Continue digestion for 2 hours to obtain the digested sample.

[0067] Table 3 Particle size and potential of samples of Examples 1 to 4 and Comparative Example 1 after digestion

[0068] After digestion Particle size (nm) Potential (mv) Example 1 711.1±2.1 -35.28±5.5 Example 2 654.2±7.9 -36.85±2.7 Example 3 597.4±3.6 -34.13±4.1 Example 4 890.6±11.4 -45.07±3.4 Comparative Example 1 1018.9±8.4 -20.69±6.9

[0069] like Figure 2 As shown, different digested β-carotene protein nano solutions have different potentials, and the absolute values ​​thereof are ranked from large to small as follows: Example 4> Example 2> Example 1> Example 3> Comparative Example 1.

[0070] Comparative Example 2

[0071] The preparation method of β-carotene corn oil emulsion comprises the following steps:

[0072] 0.1% mass volume fraction of β-carotene was dissolved in corn oil, and stirred at 50° C. for 30 minutes using magnetic stirring to ensure that the β-carotene was completely dissolved. The obtained sample was the β-carotene corn oil emulsion.

[0073] Example 7

[0074] Cytotoxicity test (MTT), including the following steps:

[0075] The cytotoxicity of the protein nanoparticles before and after digestion was determined by the MTT method. A 5 mg / mL MTT solution was prepared with PBS solution. 100 μL of 5.0×10 5 / mL of cell solution, incubate at 37℃ for about 12h, so that the cell confluence of each well reaches about 40% for subsequent experiments. First, wash the cells twice with PBS solution, then add samples with different dilution multiples (2x, 5x, 10x, 20x, 40x), and incubate at 37℃ for 4h. Remove the culture medium, wash the cells with PBS solution, add 100μL of MTT solution, and incubate at 37℃ for 2h. Remove the culture medium, wash the cells with PBS solution, and add 100μL of dimethyl sulfoxide. Shake the 96-well plate for 5min to completely dissolve the purple crystals of MTT in DMSO. Use an enzyme reader to detect the optical density (OD value) of each well of cells at a wavelength of 490nm, and calculate the cell survival rate according to the following formula (1):

[0076]

[0077] Table 4 Cytotoxicity of Examples 1 to 4 and Comparative Example 1 at different dilution times

[0078] After digestion Dilute 2 times Dilute 5 times Dilute 10 times Dilute 20 times Dilute 40 times Example 1 64.31％±0.17 79.98％±0.13 82.67％±0.18 87.91％±0.18 90.65％±0.21 Example 2 62.74％±0.17 76.58％±0.18 80.16％±0.13 85.23％±0.15 89.49％±0.14 Example 3 58.22％±0.2 72.46％±0.11 77.96％±0.11 81.12％±0.14 86.37％±0.18 Example 4 66.84％±0.12 80.29％±0.11 87.24％±0.15 90.53％±0.17 92.96％±0.12 Comparative Example 1 69.43％±0.14 82.18％±0.14 89.29％±0.16 91.95％±0.23 94.82％±0.11 Comparative Example 2 61.56％±0.18 75.71％±0.15 79.14％±0.14 83.92％±0.16 88.15％±0.19

[0079] like Figure 3 As shown, the cell survival rate of the sample obtained in Example 1 is ≥75% when the dilution rate after digestion is greater than 5 times. The survival rate of the sample obtained in Example 2 is ≥75% when the dilution rate after digestion is greater than 5 times. The survival rate of the sample obtained in Example 3 is ≥75% when the dilution rate after digestion is greater than 10 times. The survival rate of the sample obtained in Example 4 is ≥75% when the dilution rate after digestion is greater than 5 times. The survival rate of the sample obtained in Comparative Example 1 is ≥75% when the dilution rate after digestion is greater than 5 times. The survival rate of the sample obtained in Comparative Example 2 is ≥75% when the dilution rate after digestion is greater than 5 times.

[0080] Example 8

[0081] The investigation of the transport and absorption of β-carotene includes the following steps:

[0082] The transport and absorption of bile salt-modified β-carotene protein nanosolution after digestion in the cell model was investigated using a Caco-2 / HT29 co-culture cell model with growth differentiation of 21 to 28 months. The method is as follows: wash the AP end cells twice with DMEM solution, then add 1.5 mL of the sample diluted with DMEM to the AP end, and add 2 mL of DMEM culture medium to the BL end. Incubate at 37°C, during which the resistance between the AP end and the BL end is regularly tested using the transmembrane resistance to ensure that the cell membrane can maintain integrity during the entire incubation process. After incubation for 4 hours, the experiment was terminated using a 4°C PBS solution containing 10% ethanol, and the DMEM solution and cells at the AP end and BL end were collected. The content of β-carotene and its metabolites was quantitatively determined using high-performance liquid chromatography.

[0083] Table 5 Nutrient content transported to cells in samples of Examples 1 to 4 and Comparative Example 1

[0084]

[0085] like Figure 4 , Figure 5 , Figure 6 As shown, the intracellular accumulation of BC and its metabolites after digestion of the samples obtained in Examples 1-4 and Comparative Examples 1 and 2 are as follows: Example 3 > Example 2 > Example 1 > Example 4 > Comparative Example 1 > Comparative Example 2.

[0086] Example 9

[0087] The properties of the finished products obtained in Examples 1 to 4 and Comparative Example 1 were measured, and the results are shown in the table. The VA liver tissue content and VA adipose tissue content were measured respectively, and the measurement results are recorded in the table.

[0088] The VA liver tissue content and VA adipose tissue content were determined as follows: mice were gavaged with samples of β-carotene protein nano-solution dispersions modified with different bile salts. Mice were gavaged with PBS as a blank control. After 30 minutes, the mice were anesthetized and their livers and epididymal fat were taken. The tissues were homogenized in 0.9% sodium chloride solution, and then a mixed organic solvent of methanol and n-hexane (containing 0.1% BHT) was added in a ratio of 2:3. The mixture was vortexed for 30 seconds and the upper organic phase was collected after centrifugation at 2000g for 4 minutes. The extraction process was repeated three times. Thereafter, the organic solvent was dried with nitrogen and redissolved in 200uL dichloromethane for HPLC quantitative analysis.

[0089] HPLC quantitative analysis method: Alliance HPLC system equipped with Waters 2998 PDA detector was used to quantitatively determine the content of β-carotene (BC) and its metabolites retinol (ROL) and retinyl palmitate (RP) in the samples. The chromatographic column was a COSMOSIL Cholester column with a precolumn (250×4.6 mm, 5 μm in diameter). HPLC chromatographic conditions are: flow rate: 1mL / min; column temperature: 10°C; sample injection volume: 20μL; detection wavelength: 450nm (for BC) and 340nm (for ROL and RP); analysis time: 28min; samples are placed at 4°C; mobile phase: A phase (84% methanol: 14% acetonitrile: 2% ultrapure water, v / v / v) and B phase (dichloromethane) mixed; gradient elution program: 0-15min, linear change from 80%A, 20%B to 45%A, 55%B; 15-20min, 45%A, 55%B conditions maintained stable; 20-25min, linear change from 45%A, 55%B to 80%A, 20%B; 25-28min, maintain 80%A, 20%B ratio stable for 3min. The standard is linear in the range of 0.1-100μg / mL. The regression equation of the calibration curve was calculated based on the peak area versus standard concentration, with R2>0.99, and the obtained data were recorded in the table.

[0090] Table 6 Accumulation of samples of Examples 1 to 4 and Comparative Example 1 in the liver and adipose tissue

[0091]

[0092] like Figure 7 As shown, the VA content in the samples obtained in Examples 1-4 and Comparative Examples 1 and 2 after digestion and transported to the liver tissue is, from large to small, Example 3> Example 2> Example 1> Example 4> Comparative Example 1> Comparative Example 2. The VA content in the adipose tissue is, from large to small, Example 3> Example 2> Example 1> Example 4> Comparative Example 1> Comparative Example 2.

[0093] The sample prepared in Example 3 has the best VA bioavailability, so sodium chenodeoxycholate can be used for modification to improve the bioavailability of carotene encapsulated in protein nanocarriers. Compared with Comparative Example 1, the bioavailability of the samples obtained in the four embodiments is better than that in Comparative Example 1, and the bioavailability of the sample obtained in Comparative Example 2 is worse than that in Comparative Example 1. Therefore, protein nano solution encapsulation and bile salt modification help to improve the effect of nutrient delivery of protein nano solution, and sodium chenodeoxycholate has the best improvement effect, followed by sodium deoxycholate. Since sodium chenodeoxycholate has a high cytotoxicity, sodium deoxycholate can also be used to improve the delivery effect of β-carotene protein nano solution.

[0094] Comparative Example 3

[0095] The difference between this comparative example and Example 1 is that the soy protein is adjusted to sodium caseinate, and the rest of the preparation process is the same as that of Example 1.

[0096] Comparative Example 4

[0097] The difference between this comparative example and Example 1 is that ethyl acetate is replaced with methanol, and the rest of the preparation process is the same as that of Example 1.

[0098] Table 7 Accumulation of samples of comparative examples 3 and 4 in liver and adipose tissue

[0099] Example VA liver tissue content (ng / g tissue) VA adipose tissue content (ng / g tissue) Comparative Example 3 1135±179 125±34 Comparative Example 4 647±106 72±29

[0100] The protein nano solution prepared in Comparative Example 3 is a casein nano solution. Casein is less resistant to digestion than soy protein, and more β-carotene is lost after digestion, so the VA liver tissue content and adipose tissue content are lower.

[0101] In Comparative Example 4, methanol was used to prepare the organic phase. Since methanol would demulsify the protein nano-solution, a large amount of β-carotene was lost.

[0102] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A method for preparing a bile salt-modified β-carotene protein nano solution, characterized in that: include, Carotene was dissolved in ethyl acetate as the organic phase; The protein was dissolved in ultrapure water as the aqueous phase; The organic phase and the aqueous phase are mixed, sheared, dispersed, homogenized, and rotary evaporated to obtain a β-carotene protein nano solution; dissolving bile salt in ultrapure water and stirring to obtain a bile salt aqueous solution; The bile salt aqueous solution and the beta-carotene protein nano solution are mixed and stirred to obtain the bile salt modified beta-carotene protein nano solution.

2. The preparation method according to claim 1, characterized in that: The bile salts include one or more of sodium cholate, sodium deoxycholate, sodium chenodeoxycholate and naturally extracted pig bile salts.

3. The preparation method according to claim 2, characterized in that: The concentration of the bile salt aqueous solution is 8 mg / mL.

4. The preparation method according to claim 3, characterized in that: The bile salt aqueous solution and the beta-carotene protein nano solution are added in a ratio of 1-2:2-4.

5. The preparation method according to claim 1, characterized in that: The added amount ratio of beta-carotene to ethyl acetate in the organic phase is 1-10:1-100.

6. The preparation method according to claim 1, characterized in that: The protein comprises one or more of soy protein, pea protein, rice protein, peanut protein, whey protein isolate, brown rice protein and oat protein.

7. The preparation method according to claim 1, characterized in that: The homogenization condition is to homogenize once at 100 MPa using a microfluidizer.

8. The preparation method according to claim 5, characterized in that: The volume ratio of the organic phase to the aqueous phase is 1:2-10.

9. The bile salt-modified β-carotene protein nano solution prepared by the preparation method according to claims 1 to 8.

10. Use of the bile salt-modified β-carotene protein nano solution according to claim 9 in oral β-carotene delivery.

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