Gastrointestinal delivery carrier of high-activity rhodotorula compound and application of gastrointestinal delivery carrier in colon fermentation

Abstract

The invention discloses a gastrointestinal delivery carrier of a high-activity rhodotorula compound and application of the gastrointestinal delivery carrier in colon fermentation, and belongs to the field of research of food science, pharmaceutical preparations, nutriology and microbiology. The invention explores the storage stability and digestive tract metabolism conditions of the rhodotorula high-activity compound under different carrier forms, namely corn oil (F-oil) loaded with the rhodotorula high-activity compound, corn oil nano-emulsion spray drying powder (F-EP) of the rhodotorula high-activity compound and corn oil nano-emulsion alginate gel (F-EPA) of the rhodotorula high-activity compound. Wherein the storage stability of the F-EPA is optimal. The three carriers do not release the high-activity rhodotorula compound in the in-vitro oral cavity and stomach stages, but in the small intestine stage, the digestibility and the biological accessibility of F-EP are the highest, and the release of F-EPA is the minimum. The F-EPA can effectively promote the rhodotorula red pigment to regulate the structural composition and abundance value of intestinal microbiome.

Description

Technical Field

[0001] The present invention relates to a gastrointestinal delivery carrier of a red yeast high-activity complex and its application in colonic fermentation, with an emphasis on improving its stability and metabolic release in the gastrointestinal tract, while also covering its application in an in vitro simulated continuous colonic fermentation process, and belongs to the research fields of food science, pharmaceutical preparations, nutrition and microbiology. Background Art

[0002] Monascus purpureus is a natural pigment produced by fermentation of red yeast. In practical applications, it is prone to oxidation during storage, has poor stability, and has low bioavailability in the gastrointestinal tract. Researchers have explored a variety of technologies to prepare red yeast purpureus nanoparticle delivery systems to solve the above problems. For example, for the small intestine, the existing technology loads red yeast purpureus into liposomes, but after oral administration, the double-layer structure of the liposome will be destroyed by intestinal bile salts and enzymes, resulting in premature release of the embedded macromolecules and reduced residence time.

[0003] Different sustained-release systems have also been developed in the prior art for the colon, including micelle loading, emulsion, nanoemulsion, nanoliposome and hydrogel. However, there are still various problems with different sustained-release systems, such as micelle loading, a technology that absorbs fat-soluble molecules on the surface of micelles (Qian J, Guo Y, Xu Y, et al. Combination of micelles and liposomes as a promising drug delivery system: a review. Drug Deliv Transl Res. 2023 Nov; 13 (11): 2767-2789.), but it has disadvantages such as low loading capacity, premature drug release and poor stability. In addition, some researchers have used water-in-oil nanoemulsions and microemulsions to encapsulate and transport lipid substances (Pavoni L, Perinelli DR, Bonacucina G, et al. An Overview of Micro-and Nanoemulsions as Vehicles for Essential Oils: Formulation, Preparation and Stability. Nanomaterials (Basel). 2020 Jan 12; 10(1): 135.), but they have the characteristics of complex preparation methods (most of which involve organic solvents), poor loading capacity and instability during storage. For example, the common nanohydrogel technology, a three-dimensional soft gel formed by cross-linking water-soluble materials (Akram, Muhammad, and Rafaqat Hussain. "Nanohydrogels: History, development, and applications in drug delivery." Nanocellulose and Nanohydrogel Matrices: Biotechnological and Biomedical Applications (2017): 297-330.), although it has great potential in promoting the effective delivery of carotenoids, it also has some limitations including poor loading capacity, premature release and oxidation of carotenoids.

[0004] The release rate of red yeast rhodotorula in the red yeast rhodotorula electrospun microspheres prepared by patent CN115399471 B is not high in the small intestine, but is higher in the colon, so it is mainly used for targeted colon delivery; the release rate of red yeast rhodotorula in the bile system of red yeast rhodotorula prepared by patent CN115226898 B in the small intestine is also not high. Although it has a certain sustained-release effect, it takes a long time for red yeast rhodotorula to achieve full release during actual use.

[0005] The study (Li Xingming. Research on the mechanism of red yeast erythrin intervention in high-fat obesity [D]. Jiangnan University, 2022.) prepared three red yeast erythrin carrier systems and explored the storage stability and metabolism of red yeast erythrin in the three carriers in the digestive tract; the study (B.Hu, C.Liu, W.Jiang, et al, Chronic in vitro fermentation and invivo metabolism: Extracellular polysaccharides from Sporidiobolus pararoseusregulate the intestinal microbiome of humans and mice, International Journal of Biological Macromolecules 192 (2021) 398-406.) found that red yeast polysaccharides extracted from red yeast have significant intestinal prebiotic properties. However, no research has confirmed whether the combination of red yeast polysaccharides and red yeast erythrin will synergistically promote the stable release of red yeast erythrin after encapsulation of the carrier and enhance the function of the intestinal environment. Summary of the invention

[0006] In order to solve the above problems, the present invention obtains a red yeast high-activity complex by mixing red yeast red yeast and red yeast polysaccharide, and then successively prepares three different red yeast high-activity complex carriers, namely, vegetable oil loaded with red yeast high-activity complex, red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder, and red yeast high-activity complex vegetable oil nanoemulsion alginate gel, to improve the stability of red yeast red yeast and promote the metabolic function of the gastrointestinal tract. Among them, the vegetable oil loaded with the red yeast high-activity complex has the best effect in improving the stability of red yeast red yeast, and the red yeast high-activity complex vegetable oil nanoemulsion alginate gel has the best effect in promoting the metabolic function of the gastrointestinal tract.

[0007] The first object of the present invention is to provide a method for preparing a red yeast highly active complex carrier, wherein the carrier is a vegetable oil loaded with a red yeast highly active complex, comprising the following steps:

[0008] (1) mixing red yeast red pigment and red yeast polysaccharide to obtain a red yeast high-activity complex;

[0009] (2) dissolving the red yeast highly active complex obtained in step (1) in vegetable oil to obtain vegetable oil loaded with the red yeast highly active complex.

[0010] In one embodiment, in step (1), red yeast rhodotorula erythrophylla and red yeast polysaccharide are mixed in a mass ratio of (8-12):1.

[0011] In one embodiment, in step (2), the vegetable oil includes but is not limited to corn oil, soybean oil, and sunflower oil.

[0012] In one embodiment, in step (2), the mass ratio of the red yeast high-activity complex to the vegetable oil is (3-5):1000.

[0013] In one embodiment, in step (1), the method for preparing rhodotorula erythrophylla comprises the following steps:

[0014] Inoculate Sporidiobolus pararoseus JD-2 into the culture medium at an inoculation ratio (bacterial liquid / culture medium volume ratio) of 2% to 5% to obtain red yeast fermentation liquid;

[0015] Centrifuge the red yeast fermentation liquid, discard the supernatant, and add water to the precipitate to obtain a yeast suspension;

[0016] The yeast suspension is homogenized and centrifuged, the supernatant is taken and then washed, the precipitate is taken, the precipitate is extracted, and vacuum dried to obtain red yeast rhodopsin.

[0017] In one embodiment, the culturing is carried out at a temperature of 25 to 30° C. and a pH value of 5.5 to 6.5.

[0018] In one embodiment, the red yeast fermentation liquid is centrifuged at 6500-7500 r / min for 15-20 min, the supernatant is discarded, and the precipitate is added with water to obtain a yeast suspension.

[0019] In one embodiment, vacuum drying is performed to obtain rhodotorula rhodotorula in the form of a lyophilized powder.

[0020] In one embodiment, the yeast suspension is homogenized at 60-100 MPa for 2-4 times, centrifuged at 4000-6000 r / min for 5-15 min, the supernatant is taken and then washed 2-3 times, the precipitate is taken, the precipitate is extracted for 20-30 min and then vacuum dried to obtain the red yeast rhodotorula lyophilized powder.

[0021] In one embodiment, in step (1), the method for preparing red yeast polysaccharide comprises the following steps:

[0022] Inoculate Sporidiobolus pararoseus JD-2 into the culture medium at an inoculation ratio (bacterial liquid / culture medium volume ratio) of 2% to 5% to obtain red yeast fermentation liquid;

[0023] Centrifuge the red yeast fermentation broth and take the precipitate;

[0024] The precipitate is extracted, centrifuged after the extraction, the supernatant is taken and then deproteinized, the centrifugation is continued, the supernatant is taken, and the mixture is allowed to stand. After the standing period is completed, the precipitate is taken and freeze-dried to obtain freeze-dried red yeast polysaccharide powder.

[0025] In one embodiment, the red yeast fermentation broth is centrifuged at 6500-7500 r / min for 15-20 min and then the precipitate is collected.

[0026] In one embodiment, freeze drying is performed to obtain Rhodozymosan in the form of a lyophilized powder.

[0027] In one embodiment, the precipitate is extracted, and after the extraction is completed, the precipitate is centrifuged at 6000-7000 r / min for 5-15 min, the supernatant is taken and the supernatant is deproteinized, and the supernatant is further centrifuged at 6000-7000 r / min for 5-15 min, the supernatant is taken, and the mixture is allowed to stand. After the standing is completed, the mixture is centrifuged at 6000-7000 r / min for 5-15 min, the precipitate is taken, and the precipitate is freeze-dried to obtain a freeze-dried red yeast polysaccharide powder.

[0028] The second object of the present invention is to provide a method for preparing a red yeast high-activity complex carrier, wherein the carrier is a red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder, comprising the following steps:

[0029] (1) mixing the OSA starch solution with the vegetable oil loaded with the red yeast highly active complex of the present invention, and homogenizing to obtain a red yeast highly active complex vegetable oil nanoemulsion;

[0030] (2) spray-drying the red yeast high-activity complex vegetable oil nanoemulsion obtained in step (1) to obtain red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder.

[0031] In one embodiment, in step (1), the OSA starch solution and the vegetable oil loaded with the red yeast highly active complex are mixed in a mass ratio of 1:(8-10).

[0032] In one embodiment, in step (1), the mass fraction of the OSA starch solution is 30-50%.

[0033] In one embodiment, in step (1), an OSA starch solution having a mass fraction of 30 to 50% is mixed with a vegetable oil loaded with a red yeast highly active complex, and stirred to obtain an initial emulsion. The initial emulsion is homogenized 4 to 8 times by a high-pressure homogenizer at 15 to 25° C. and 90 to 110 MPa to obtain a red yeast highly active complex vegetable oil nanoemulsion.

[0034] In one embodiment, in step (2), the operating conditions of the spray drying are: an air inlet temperature of 180-200°C, an air outlet temperature of 80-90°C, a feed flow rate of 1-3 L / h, a nozzle atomization pressure of 2-4 bar, and a duration of the spray drying process of 1-3 h.

[0035] The third object of the present invention is to provide a method for preparing a red yeast high-activity complex carrier, wherein the carrier is a red yeast high-activity complex plant oil nanoemulsion alginate gel, comprising the following steps:

[0036] The alginate solution is mixed with the red yeast high-activity complex vegetable oil nanoemulsion of the invention, and gelled to obtain the red yeast high-activity complex vegetable oil nanoemulsion alginate gel.

[0037] In one embodiment, alginate solution and red yeast high-activity complex plant oil nanoemulsion are mixed in a mass ratio of (1-2):(1-2), homogenized, and stirred to form a gel block to obtain red yeast high-activity complex plant oil nanoemulsion alginate gel.

[0038] In one embodiment, the gel block is formed by mixing alginate solution and red yeast high-activity complex plant oil nanoemulsion in a mass ratio of (1-2): (1-2), homogenizing, maintaining a stirring speed of 200-400 rpm, continuing for 3-10 minutes, and reacting at 30-40° C. for 5-15 minutes.

[0039] In one embodiment, the alginate solution includes but is not limited to sodium alginate solution, potassium alginate solution, calcium alginate solution, preferably a sodium alginate solution with a mass concentration of 2%.

[0040] In one embodiment, a cross-linking agent having a mass concentration of 1 to 3% is used for gelation to obtain a red yeast high-activity complex plant oil nanoemulsion alginate gel.

[0041] In one embodiment, the cross-linking agent includes but is not limited to CaCl2, propylene diamine, polyethylene glycol, and polypropylene glycol glycidyl ether.

[0042] The fourth object of the present invention is to use the vegetable oil loaded with red yeast high-activity complex, the red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder, and the red yeast high-activity complex vegetable oil nanoemulsion alginate gel prepared by the present invention in improving the stability of red yeast rhodotorula.

[0043] The fifth objective of the present invention is to use the vegetable oil loaded with red yeast high-activity complex, the red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder, and the red yeast high-activity complex vegetable oil nanoemulsion alginate gel prepared by the present invention in the preparation of products that promote gastrointestinal metabolic function.

[0044] Beneficial Effects

[0045] (1) The present invention prepares a novel red yeast high-activity complex and applies it to a carrier packaging delivery system, which can significantly improve the stability of red yeast rhodotorula. Corn oil (F-oil) loaded with the red yeast high-activity complex as a carrier has excellent storage stability; the red yeast high-activity complex corn oil nanoemulsion alginate gel (F-EPA) has a better sustained-release effect and can effectively delay the release of red yeast rhodotorula, so that it can be stably present in the gastrointestinal tract for a longer time;

[0046] (2) The present invention uses alginate as an emulsifier and combines it with OSA starch to prepare a nanoemulsion of a red yeast high-activity complex, and encapsulates it to prepare a new red yeast high-activity complex carrier form (F-EPA), which significantly improves the stability of red yeast erythrin during storage and gastrointestinal transport, making the encapsulation process simple and efficient;

[0047] (3) The delivery effect of the red yeast highly active complex in the digestive tract in the form of F-EPA is better than other carriers. It can effectively release the red yeast erythrin, which is intolerant to gastric acid and has poor stability, into the colon, ensuring the stable presence of red yeast erythrin in the gastrointestinal tract and exerting healthy functions in the colon through interaction with the microbiota;

[0048] (4) In an in vitro simulated continuous colonic fermentation system, F-EPA significantly increased the concentrations of propionic acid, butyric acid, and valeric acid, and effectively regulated the composition and abundance of intestinal flora. This effect indicates that this carrier form can directly or indirectly promote the metabolism of red yeast rhodotorula in the human intestinal flora, increase the production of beneficial short-chain fatty acids, and improve intestinal health. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 It is a flow chart of the overall idea of ​​the present invention.

[0050] Figure 2The present invention discloses the changes of short-chain fatty acids (SCFAs) in the in vitro fermentation of the red yeast highly active complex corn oil nanoemulsion alginate gel (F-EPA) and the red yeast red pigment corn oil nanoemulsion alginate gel (T-EPA) of the present invention, and their effects on phylum-level species and genus-level species in the human intestinal microbiome; wherein, (A) and (D) are respectively the changes of SCFAs by F-EPA and T-EPA; (B) and (E) are respectively the effects of F-EPA and T-EPA on phylum-level species in the human intestinal microbiome; (C) and (F) are respectively the effects of F-EPA and T-EPA on genus-level species in the human intestinal microbiome. DETAILED DESCRIPTION

[0051] The present invention is further described below in conjunction with examples, but the embodiments of the present invention are not limited thereto.

[0052] 1. Raw materials used in the examples and comparative examples

[0053] Corn oil, soybean oil, and sunflower oil were purchased from the Xuelang District Farmers’ Market in Wuxi;

[0054] OSA modified starch was provided by Huahao Huafeng Starch Co., Ltd.;

[0055] Lipase, mucin, pepsin, bile extract, and Nile red were purchased from Sigma, USA;

[0056] Standard products of acetic acid, propionic acid, isobutyric acid, n-butyric acid, isovaleric acid and n-valeric acid were purchased from Sigma-Aldrich Chemical (St. Louis, Missouri, USA); the remaining reagents were of analytical grade and purchased from Tianjin Kaitong Chemical Reagent Co., Ltd.

[0057] The Sporidiobolus pararoseus JD-2 involved in the following examples is recorded in the Chinese invention patent with publication number CN102115716B and deposit number CCTCC M 2010326.

[0058] 2. Physical and chemical properties of different carriers

[0059] (1) Determination of particle size and zeta potential of emulsions with different carriers

[0060] Corn oil loaded with red yeast highly active complex (F-oil) and red yeast highly active complex corn oil nanoemulsion spray-dried powder (F-EP) were dissolved in ultrapure water at a dilution ratio of 1:1000, and the particle size and charge (ζ potential) of the emulsion were measured using a nanoparticle size and ζ potential instrument.

[0061] For the gel particles of F-EPA, the diameters of at least 20 particles were measured using a digital micrometer (0-300 mm, EC10, high-precision digital caliper), and the average and standard deviation thereof were calculated.

[0062] The particle size and zeta potential of different carriers of red yeast rhodotorula were also measured according to the above method.

[0063] (2) Encapsulation yield and efficiency of red yeast rhodotorula in different carriers

[0064] The role of encapsulation yield (EY) and encapsulation efficiency (EE):

[0065] For red yeast high-activity complex: used to evaluate the retention of red yeast rhodopsin in red yeast high-activity complex in different carrier forms;

[0066] For red yeast rhodotorula: used to evaluate the retention of red yeast rhodotorula in different carrier forms.

[0067] Encapsulation yield (EY):

[0068] For the red yeast high-activity complex: EY is the ratio of the amount of red yeast rhodopsin in the final red yeast high-activity complex to the total amount of red yeast rhodopsin in the red yeast high-activity complex initially added;

[0069] For red yeast rhodotorula: EY is the ratio of the final amount of red yeast rhodotorula to the total amount of red yeast rhodotorula initially added.

[0070] Encapsulation efficiency (EE):

[0071] For the red yeast high-activity complex: EE is the ratio of the content of red yeast rhodopsin removed from the surface of the prepared red yeast high-activity complex embedded sample to the content of the total red yeast rhodopsin in the prepared red yeast high-activity complex embedded sample;

[0072] For rhodotorula erythrin: EE is the ratio of the content of rhodotorula erythrin in the embedded sample excluding the surface rhodotorula erythrin to the total rhodotorula erythrin in the embedded sample.

[0073] (3) Storage stability determination of rhodotorula rubrum in different carriers

[0074] 2.0mL corn oil loaded with red yeast high-activity complex (F-oil), 0.15g red yeast high-activity complex corn oil nanoemulsion spray-dried powder (F-EP) and 0.25g red yeast high-activity complex corn oil nanoemulsion alginate gel (F-EPA) were placed in 10mL brown bottles and stored in the dark at 50℃, 25℃ and 4℃, while the same amount of samples were placed in transparent glass bottles for light storage. The concentration and changes of the main component (red yeast erythrin) in the red yeast high-activity complex samples were measured at fixed time intervals.

[0075] The calculation formula of retention rate is C t / C1, where C1 is the red yeast red yeast concentration measured before storage of the red yeast high-activity complex, C t is the concentration of red yeast rhodotorula in the red yeast high-activity complex at storage time t.

[0076] The storage stability of different carriers of red yeast red pigment and other carrier forms of red yeast high-activity complexes were also determined according to the above method.

[0077] 3. Simulate the ascending-transverse-descending colon fermentation process in vitro to study the metabolism of the red yeast highly active complex in F-EPA and the red yeast rhodopsin in T-EPA. The specific steps are as follows:

[0078] Fecal sample processing: Feces from 8 healthy young people were collected, mixed with sterile PBS (pH=6.8) (ratio 1:80), stirred and filtered to obtain intestinal bacterial fluid, which was placed in an anaerobic environment.

[0079] Fermentation setup: The bacterial liquid was inoculated into a fermenter containing 10% culture medium, the pH was adjusted to 7.0, and the fermentation was carried out at 37°C and 150 rpm.

[0080] Add samples: F-EPA and T-EPA (1% by mass) were added to the fermenter, 30 mg was added every day, and the fermentation was continued for 2 days.

[0081] Sample collection and analysis: Fermentation broth was collected regularly during 0-48 h for metabolic analysis.

[0082] 4. Digestion characteristics and bioaccessibility of red yeast high-activity complex and red yeast rhodotorula in different carriers

[0083] (1) In vitro simulation of oral, gastric, and small intestinal (GIT) digestion

[0084] Study the digestion characteristics of the sample. The steps are as follows:

[0085] Oral stage: 5 mL of sample was mixed with 4 mL of simulated oral fluid (1.408 g KCl, 1.428 g NaHCO3, 0.629 g KH2PO4, 0.038 g MgCl2 6H2O, 0.007 g (NH4)2CO3 dissolved in 1 L deionized water) and incubated at 37°C. 25 μL 0.3 mol / L CaCl2 and 0.975 mL water were added, mixed and incubated in a 37°C water bath for 2 minutes.

[0086] Gastric stage: Mix the oral stage mixture with 8 mL of simulated gastric fluid (0.643 g KCl, 2.625 g NaHCO3, 0.153 g KH2PO4, 0.031 g MgCl2·6H2O, 0.060 g (NH4)2CO3, 2.808 g NaCl dissolved in 1 L deionized water), add 5 μL 0.3 mol / L CaCl2, 4 mg gastric lipase (60 U / mL), 400 mg pepsin (2000 U / mL), and 1.895 mL H2O, adjust the pH to 3.0, and incubate in a 37°C water bath.

[0087] Small intestine stage: Mix the gastric stage mixture with 8.5 mL of simulated intestinal fluid (3.452 g NaCl, 0.634 g KCl, 8.952 g NaHCO3, 0.136 g KH2PO4, 0.084 g MgCl2·6H2O dissolved in 1 L deionized water), add 40 μL 0.30 mol / LCaCl2, 16.00 mg trypsin (100 U / mL), 2.67 mg pancreatic lipase (2000 U / mL), 272.38 mg bile salt (10 mmol / L), and 11.26 mL H2O, mix well, adjust the pH to 7.0, and incubate in a 37°C water bath.

[0088] The free fatty acids (FFAs) released by lipid digestion in the sample can be calculated based on the volume of alkali solution consumed to neutralize the sample (pH 7.0) during simulated small intestinal digestion using the following formula:

[0089] Total free fatty acid release (%) = V NaOH(t) ×C NaOH ×M w,lipid / 2m lipid ×100%

[0090] In the formula, m lipid is the total mass of fat in the simulated digestive fluid (g), V NaOH(t) is the volume of NaOH added to the reaction vessel when the sample is in the simulated small intestine digestion time t, C NaOH is the concentration of NaOH in the burette (0.25 M), M w,lipid Average molecular weight of lipids (corn oil 872 g / mol).

[0091] (2) Release rate of red yeast rhodotorula in different carriers

[0092] After oral and gastric digestion, the digestion products coated with different carrier forms reach the small intestine. In this experiment, samples were taken at different digestion time points in the small intestine, and the concentration of red yeast rhodotorula in the dissolution solution was determined by high performance liquid chromatography (HPLC) and ultraviolet visible spectrophotometry (UV-Vis). Ultraviolet spectroscopy is usually used to determine the content of red yeast rhodotorula, and red yeast rhodotorula usually has an absorption peak near 480nm at a specific wavelength.

[0093] Calculate the release amount at each time point:

[0094] Based on the concentration of rhodotorula rhodotorula in the dissolution solution and the sampling volume, the amount of rhodotorula rhodotorula rhodotorula released at each time point can be calculated: M(t) = C(t) × V;

[0095] Wherein, M(t) is the mass of rhodotorula rhodotorula released at time t (unit: mg or μg); C(t) is the concentration of rhodotorula rhodotorula in the dissolution solution at time t (unit: mg / mL or μg / mL); V is the volume of each sampling (unit: mL).

[0096] Calculate the total release rate:

[0097] The total release rate refers to the ratio of the amount of rhodotorula rhodotorula released in the dissolution solution to the initial total amount at a certain time point or within a certain time interval. The release rate R(t) at each time point can be calculated as: R(t) = M(t)M0 × 100% R(t);

[0098] Wherein, R(t) is the release rate (%) at time t; M(t) is the mass of rhodotorula ...

[0099] The release rates of different carriers of red yeast red pigment and other carrier forms of red yeast high-activity complex were also determined according to the above method.

[0100] (3) Bioaccessibility of Rhodotorula rhodotorula in different carriers

[0101] After the small intestinal digestion stage, an equal amount of chyme (10000 g) was collected and centrifuged at 4°C for 40 minutes; then, the centrifuged chyme (0.1 g) was dispersed in deionized water (0.9 mL), 0.1 mL of the solution was mixed with 0.9 mL of DMSO, and 2.0 mL of n-hexane / dichloromethane (4:1) was used to extract red yeast rhodotorula; after centrifugation (3000 rpm, 10 min), the red yeast rhodotorula content in the organic phase was determined by high performance liquid chromatography, and the red yeast rhodotorula content in the supernatant was detected by the above method. The calculation formula for bioaccessibility is:

[0102] Bioaccessibility (%) = (C Micelle / C Digesta )×100%;

[0103] In the formula, C Micelle is the concentration of red yeast rhodotorula in the digestion supernatant, C Digesta It is the concentration of rhodotorula rhodotorula in the chyme at the end of digestion.

[0104] 5. Distribution of red yeast highly active complex in F-EPA and red yeast rhodotorula in T-EPA and evaluation of their efficacy in regulating the intestinal environment

[0105] (1) Detection of short-chain fatty acids (SCFAs) content

[0106] According to the in vivo research results (Liu C, Li X, Li J, et al. Role of the gut microbiota indietary patterns rich in torularhodin via OSA colon-targeted delivery. Food Funct. 2022 Oct 31; 13(21): 11034-11048.), the SCFAs content of F-EPA samples was further detected:

[0107] 200 mg of red yeast high-activity composite corn oil nanoemulsion alginate gel (F-EPA) sample was placed in a 2 mL centrifuge tube, 0.1 mL of KHSO4 saturated solution and 0.1 mL of 2-ethylbutyric acid internal standard solution (0.1 mg / mL) were added, and 1 mL of dimethyl carbonate (DMC) was added, and the tube was vigorously shaken and sealed for 10 minutes;

[0108] The mixture was then centrifuged at 3800 r / min for 10 min to separate the solid phase and aqueous phase from the DMC upper layer, and the DMC phase was pipetted into a gas phase vial;

[0109] A mixed SCFAs standard solution (including formic acid, acetic acid, propionic acid, n-butyric acid, isobutyric acid, n-valeric acid and isovaleric acid) was prepared at 0.1 mg / mL, 0.5 mg / mL, 0.75 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL and 3.0 mg / mL, and pre-treated according to the above method.

[0110] SCFAs were detected using the GC-MS method with EI mode, emission current 1 mA, electron energy 70 eV, interface temperature 250 °C, source temperature 210 °C, voltage 2000 V, initial temperature 60 °C, initial time 1 min, heating rate 15 °C / min, final temperature 240 °C, and holding time 10 min.

[0111] (2) 16S rRNA gene sequencing

[0112] Use a DNA extraction kit to extract sample DNA and perform electrophoresis detection;

[0113] The V3-V4 region of the sample 16S rRNA was amplified by PCR using primers. The PCR amplification products were detected by 2% agarose gel electrophoresis and recovered using the AMPure XT beads kit;

[0114] The purified PCR products were quantitatively tested using Qubit, and qualified sequencing libraries were sequenced using the Illumina MiSeq sequencer for PE300 sequencing;

[0115] The operational taxonomic units (OTUs) table of microbial sequencing clustering was screened using QIIME2 software, and the results were stored in the Greengene database;

[0116] The diversity of the microbial composition of the samples was analyzed using R and other software.

[0117] Red yeast erythrogen corn oil nanoemulsion alginate gel (T-EPA) regulates the intestinal environment and is also measured according to the above method.

[0118] Example 1: Preparation of corn oil (F-oil) loaded with red yeast high activity complex

[0119] The specific implementation includes the following steps:

[0120] 1. Preparation of raw materials

[0121] (1) Preparation of lyophilized powder of red yeast rice:

[0122] Rhodotorula rhodotorula (95%, HPLC) was isolated and purified from the extract of Sporidiobolus pararoseus JD-2 (disclosed in the Chinese invention patent CN102115716B) in our laboratory. The specific preparation method is as follows:

[0123] Sporidiobolus pararoseus JD-2 was inoculated into a medium containing glucose, yeast extract and minerals at an inoculum ratio of 3% (bacterial liquid / medium volume), and the culture was shaken at 28°C, pH 6, and a 12-hour light / 12-hour dark cycle to prepare a red yeast fermentation broth;

[0124] The red yeast fermentation liquid was centrifuged at 6500 r / min for 15 min using an ultracentrifuge, the supernatant was discarded, and the precipitate was made into a yeast suspension with a yeast mud concentration of 500 g / L using deionized water;

[0125] Subsequently, the yeast suspension was added to a high-pressure homogenizer and homogenized three times at a pressure of 80 MPa, followed by centrifugation for 10 min (5000 r / min) using a low-speed centrifuge, the supernatant was taken and washed twice with deionized water, precipitated, 100 mL of a mixture of ethanol and n-hexane (volume ratio 1:1) was added to the precipitate and extracted in the dark for 25 min to obtain a red yeast rhodopsin extract, and the red yeast rhodopsin extract was vacuum freeze-dried to obtain red yeast rhodopsin freeze-dried powder.

[0126] (2) Preparation of red yeast polysaccharide:

[0127] Rhodotorula polysaccharide (purity 83%) is also extracted and purified from the fermentation broth of the yeast strain Sporidiobolus pararoseus JD-2. The specific preparation method is as follows:

[0128] Prepare red yeast fermentation broth according to the method in step (1), then centrifuge at 6500 r / min for 15 min using an ultracentrifuge, discard the supernatant, and take the precipitate;

[0129] The precipitate was subjected to hot water extraction (the hot water extraction conditions were set as follows: extraction time was 3 h, temperature was 80 °C, and the solid-liquid ratio of precipitate to hot water was 1:20). After the extraction, the mixture was centrifuged at 6500 r / min for 10 min, and the supernatant was taken.

[0130] The supernatant was deproteinized by zinc acetate-potassium ferrocyanide method, centrifuged at 6500r / min for 10min, the supernatant was taken, 4 times the volume of anhydrous ethanol was added to the supernatant and allowed to stand for 24h. After standing, the mixture was centrifuged at 6500r / min for 10min, the precipitate was collected, and the collected precipitate was freeze-dried to obtain red yeast polysaccharide freeze-dried powder.

[0131] 2. Preparation of corn oil (F-oil) loaded with red yeast high activity complex:

[0132] The red yeast red lyophilized powder and the red yeast polysaccharide lyophilized powder obtained in step 1 are mixed at a mass ratio of 10:1, that is, 10 parts of red yeast red lyophilized powder and 1 part of red yeast polysaccharide lyophilized powder are mixed to obtain a mixture (i.e., a red yeast high-activity complex), and 0.3 g of the mixture is added to 75 mL of corn oil to obtain corn oil (F-oil) loaded with a red yeast high-activity complex with a mass fraction of 0.4%.

[0133] Example 2: Preparation of red yeast high activity composite corn oil nanoemulsion spray dried powder (F-EP)

[0134] The specific implementation includes the following steps:

[0135] (1) dispersing OSA starch in ultrapure water, stirring at 80° C. and 12,000 rpm for 30 minutes until dissolved, to obtain an OSA starch solution with a mass fraction of 40%;

[0136] (2) The F-oil obtained in Example 1 and the OSA starch solution obtained in step (1) were mixed in a mass ratio of 1:9, and stirred at 15000 rpm for 3 minutes to obtain an initial emulsion. The obtained initial emulsion was homogenized by a high-pressure homogenizer at 20° C. and 100 MPa for 6 times to obtain a red yeast high-activity composite corn oil nanoemulsion;

[0137] (3) The red yeast high-activity composite corn oil nanoemulsion obtained in step (2) was treated with Mobile Minor TM The spray dryer is used for spray drying, and the drying chamber size is controlled to be 800×620mm, 60° cone, equipped with a downstream nozzle atomizer, air inlet temperature of 190℃, outlet temperature of 85℃, feed flow rate of 2L / h, nozzle atomization pressure of 3bar, and duration of the spray drying process of 2h to obtain red yeast high-activity complex corn oil nanoemulsion spray-dried powder (F-EP). F-EP was stored in a brown glass bottle and kept away from light at 4℃ for future use.

[0138] Example 3: Preparation of red yeast high activity composite corn oil nanoemulsion alginate gel (F-EPA)

[0139] The specific implementation includes the following steps:

[0140] (1) Preparation of 2% sodium alginate:

[0141] The deionized water was heated to room temperature and stirred at 300 rpm for 1.5 hours to ensure that the sodium alginate was completely dissolved, and then the pH was adjusted to 6.5 with 0.1 M dilute hydrochloric acid. The solution was filtered through a 0.45 μm filter membrane to obtain sodium alginate with a mass concentration of 2%;

[0142] (2) The sodium alginate with a mass fraction of 2% obtained in step (1) was mixed with the red yeast high-activity composite corn oil nanoemulsion in Example 2 in a mass ratio of 1:1, and a CaCl2 liquid with a mass concentration of 2% was used as a cross-linking agent to gel the alginate (the stirring speed was maintained at 300 rpm, the stirring was continued for 5 minutes, and the reaction was carried out at 30° C. for 10 minutes to form a gel block with a size of about 5 cm×5 cm×1 cm) to obtain a red yeast high-activity composite corn oil nanoemulsion alginate gel (F-EPA), which was freeze-dried for 48 hours and then used for later use.

[0143] Comparative Example 1: Preparation of corn oil loaded with rhodotorula ...

[0144] According to the preparation method of Examples 1 to 3, the difference is that 1 part of red yeast polysaccharide freeze-dried powder is replaced by 1 part of red yeast rhodotorula erythrophore lyophilized powder (red yeast rhodotorula erythrophore lyophilized powder accounts for 11 parts, that is, 0.3g of red yeast rhodotorula erythrophore lyophilized powder), and three different carriers of red yeast rhodotorula erythrophore are prepared, namely corn oil loaded with red yeast rhodotorula erythrophore (T-oil), red yeast rhodotorula erythrophore corn oil nanoemulsion spray-dried powder (T-EP), and red yeast rhodotorula erythrophore corn oil nanoemulsion alginate gel (T-EPA).

[0145] The different carriers of Examples 1 to 3 and Comparative Example 1 were tested. The results are as follows.

[0146] 1. Physical and chemical properties testing

[0147] Table 1 Emulsion particle size, zeta potential, encapsulation efficiency (EE) and encapsulation yield of different carriers of Examples 1 to 3 and Comparative Example 1

[0148]

[0149] 2. Stability and preservation characteristics of red yeast rhodotorula

[0150] The retention rates of red yeast rhodotorula in different carriers are shown in Table 2 and Table 3:

[0151] Table 2 Average retention rate of rhodotorula erythrin in F-oil, F-EP and F-EPA of Examples 1 to 3 under light and darkness for 7 days (%)

[0152]

[0153]

[0154] Table 3 Average retention rate of rhodotorula erythrin in T-oil, T-EP and T-EPA of Comparative Example 1 for 7 days under light and darkness (%)

[0155]

[0156] It can be seen that the retention rate of the red yeast high-activity complex group was relatively higher than that of the red yeast rhodotorula group as a whole.

[0157] 3. Digestion characteristics and bioaccessibility of rhodotorula rhodotorula in different carriers

[0158] (1) In vitro simulation of oral, gastric, and small intestinal (GIT) digestion

[0159] The stability of the red yeast high-activity complex in the gastrointestinal tract is significantly affected by its oil phase digestion characteristics. As a substance dissolved in the oil phase, the digestibility and digestion degree of the red yeast high-activity complex can reflect its stability during gastrointestinal transportation, because the digestion of the oil phase will lead to the release of red yeast rhodopsin.

[0160] The three stages of digestion, oral cavity, stomach and small intestine (GIT), were simulated in vitro in a test tube. The results showed that the oil phase had little effect on digestion in the oral and gastric stages, and the main digestion occurred in the small intestine.

[0161] (2) Release rates of FFA and rhodotorula rubrum from different carriers after 2 hours of intestinal digestion

[0162] After the entire simulated GIT model, the release rates of FFA (%) and the release rates of rhodotorula rubrum in different carriers (%) were measured after 2 hours of digestion in the small intestine. The results are shown in Table 4:

[0163] Table 4 Release rate of FFA (%) and release rate of rhodozyma rhodozyma in different carriers of Examples 1 to 3 and Comparative Example 1 after 2 hours of digestion in the small intestine

[0164]

[0165] The release rate of free fatty acids (FFAs) during in vitro simulated small intestinal lipid digestion showed:

[0166] In the T-oil sample, FFAs were almost completely released within 2 hours, indicating that its oil phase was completely lipolyzed in a short time, thereby completely releasing rhodotorula rhodotorula in the small intestine; in the T-EP sample, the rate of FFA release was significantly slower, with about 60% of FFA released after 2 hours; T-oil released 40% of FFA after 2 hours. In comparison, the FFA release rates of F-oil, F-EP and F-EPA after 2 hours of small intestinal digestion were 82.3%, 71.6% and 32.9%, respectively.

[0167] The release rate of red yeast rhodotorula after 2 hours of in vitro simulation of samples prepared with different embedding was shown as follows:

[0168] The red yeast release rates of F-EPA, F-EP and F-oil were 62.9%, 92.7% and 72.3%, respectively, which were significantly lower than 67.2%, 96.5% and 84.3% of T-EPA, T-EP and T-oil.

[0169] Combined with the experimental research results of the above FFA, it is shown that the red yeast high-activity complex coated with different carriers has a stronger colon delivery ability, and the red yeast high-activity complex in the form of F-EPA carrier can significantly reduce the loss in the gastrointestinal tract in the oil-filled alginate matrix, thereby enhancing its colon targeted delivery ability.

[0170] (3) Bioaccessibility of rhodotorula rhodotorula in different carriers after 2 hours of digestion in the small intestine

[0171] Bioaccessibility refers to the proportion of the red yeast highly active complex absorbed in the gastrointestinal tract, reflecting its bioavailability. Since the red yeast highly active complex is mainly composed of red yeast rhodotorula, its bioavailability is evaluated based on rhodotorula rhodotorula. During the oil phase hydrolysis process, rhodotorula rhodotorula is released and forms micelles containing bile salts, fatty acids and phospholipids, which are absorbed by the small intestine. Therefore, its bioavailability can be measured by the concentration of rhodotorula rhodotorula in micelles during in vitro digestion.

[0172] After the entire simulated GIT model, the bioaccessibility of red yeast rhodotorula in different carriers was measured after 2 hours of digestion in the small intestine. The results are shown in Table 5:

[0173] Table 5 Bioaccessibility of rhodotorula rhodotorula in different carriers of Examples 1 to 3 and Comparative Example 1 after 2 hours of digestion in the small intestine (%)

[0174]

[0175] Among them, the bioaccessibility of F-EPA, F-EP and F-oil was lower than that of T-EPA, T-EP and T-oil, indicating that the red yeast highly active complex is more beneficial to its function in the colon. Compared with the T-series, the lower bioavailability of the F-form of the red yeast highly active complex proves its stability and potential for colon health.

[0176] 4. Test on the effects of different carriers (F-EPA, T-EPA) on regulating the intestinal environment

[0177] Based on the previous in vivo research results, the SCFAs content of F-EPA and T-EPA samples was further tested and their effects on intestinal flora were investigated:

[0178] (1) Changes in short-chain fatty acid content

[0179] The intestinal microbiota can produce short-chain fatty acids (SCFAs) by encoding various endogenous enzymes and fermenting various compounds. Studies have found that SCFAs can act on multiple local and systemic targets in the host and play an important role in the host's metabolic and immune processes.

[0180] a. Effect of red yeast high-activity complex corn oil nanoemulsion alginate gel (F-EPA) on the content of short-chain fatty acids

[0181] Depend on Figure 2 (A) It can be seen that:

[0182] The red yeast high-activity complex in F-EPA increased the total SCFAs concentration from 14.53mM (0h) to 53.24mM (48h), which was lower than the inulin content of the positive control group 61.06mM (48h), but significantly higher than the blank control group 16.79mM (48h) (p<0.05).

[0183] Acetic acid, propionic acid and butyric acid are the main fermentation products of F-EPA.

[0184] Compared with the blank control group (3.77 mM), the concentration of acetic acid in the red yeast high-activity complex increased significantly to 36.23 mM after 48 hours of fermentation (p>0.05);

[0185] The propionic acid content of F-EPA reached 9.76 mM at 48 h, significantly higher than 3.12 mM (48 h) of the control group;

[0186] Compared with the blank control group, the butyric acid concentration in the red yeast high-activity complex group was also significantly increased; while the valeric acid content showed a slow growth trend, which may be related to the metabolic activity of microorganisms.

[0187] F-EPA is metabolized into short-chain fatty acids such as acetate, propionate and butyrate under the fermentation of intestinal flora, which can protect the intestinal barrier, regulate immunity, inhibit the growth of harmful microorganisms, and promote liver fatty acid oxidation and regulate the activity of liver immune cells through the "gut-liver" axis.

[0188] Therefore, F-EPA has the potential to regulate the intestinal environment.

[0189] b. Effect of red yeast erythrin corn oil nanoemulsion alginate gel (T-EPA) on the content of short-chain fatty acids

[0190] Depend on Figure 2 (D) It can be seen that:

[0191] After 48 h of fermentation, the red yeast rhodotorula in T-EPA increased the total SCFAs concentration from 11.44 mM to 41.99 mM, the acetate concentration from 6.56 mM to 28.91 mM, the propionate concentration from 2.52 mM to 7.35 mM, and the butyrate concentration from 1.34 mM to 3.52 mM.

[0192] Therefore, the F-EPA group showed a better effect of producing short-chain fatty acids compared with the T-EPA group.

[0193] (2) Changes in intestinal flora

[0194] The human intestine contains a diverse community of symbiotic microorganisms, among which Proteobacteria, Bacteroidetes and Firmicutes are key bacteria for degrading complex macromolecules. Firmicutes / Bacteroidetes are associated with diseases such as obesity, while the metabolites of Bacteroidetes can not only be used by other bacteria, but also play an important role in improving immune system disorders.

[0195] a. Effects of red yeast high-activity complex corn oil nanoemulsion alginate gel (F-EPA) on phylum-level and genus-level species in the human gut microbiome

[0196] Figure 2 (B) Display:

[0197] At the phylum level, after 48 h of fermentation, the dominant bacterial communities in the red yeast high-activity complex group, the positive control inulin group, and the blank control OR group were Bacteroidetes, Firmicutes, and Proteobacteria.

[0198] Compared with the blank control OR and the positive control inulin group, the number of Proteobacteria in the red yeast high-activity complex group (F-EPA) was significantly reduced, while the number of Bacteroidetes was significantly increased;

[0199] Compared with the OR group, the ratio of Firmicutes / Bacteroidetes caused by F-EPA fermentation was significantly reduced, indicating that the red yeast high-activity complex has the potential to alleviate obesity symptoms caused by the imbalance between Firmicutes and Bacteroidetes.

[0200] F-EPA significantly reduced the number of Shigella coli, inhibited the proliferation of harmful bacteria, and helped prevent intestinal infections. At the same time, F-EPA promoted the proliferation of Bacteroides and improved the fermentation process ( Figure 2 (C)); while the beneficial bacteria Dialister expressing polysaccharide degradation genes decreased slightly, which may be related to its selectivity for carbohydrates.

[0201] b. Effects of red yeast EPA corn oil nanoemulsion alginate gel (T-EPA) on phylum- and genus-level species in the human gut microbiome

[0202] Depend on Figure 2 (E) Figure 2 (F) It can be seen that the microbial community of the T-EPA fermentation group mainly includes Firmicutes, Bacteroidetes, Actinobacteria and Proteobacteria, accounting for 90% of the microbial community. With the extension of fermentation time, the ratio of Firmicutes to Bacteroidetes in the T-EPA fermentation group increased significantly, and the abundance of Proteobacteria also increased significantly, indicating that red yeast rhodotorula effectively regulated the composition of intestinal microorganisms. At the genus level, the main microorganisms in the T-EPA group included Escherichia coli-Shigella, Streptococcus, Bacteroidetes, Bifidobacterium and Megasphaera.

[0203] Compared with the two, red yeast high-activity complex corn oil nanoemulsion alginate gel (F-EPA) showed a more significant regulatory effect on the composition and abundance of intestinal flora than red yeast red pigment corn oil nanoemulsion alginate gel (T-EPA), and can effectively inhibit the reproduction of harmful bacteria, thus showing potential prebiotic potential.

[0204] Comparative Example 2: Preparation and performance testing of other carrier forms of red yeast high-activity complexes (liposomes, vesicles, bile bodies, microspheres)

[0205] 1. Preparation of high-activity red yeast complexes in other carrier forms

[0206] According to the preparation method in Example 1, first prepare the red yeast red pigment freeze-dried powder and the red yeast polysaccharide freeze-dried powder, and then mix the red yeast red pigment freeze-dried powder and the red yeast polysaccharide freeze-dried powder at a mass ratio of 10:1 to obtain a red yeast high-activity complex.

[0207] The red yeast high activity complex is prepared into the following other carrier forms:

[0208] (1) Preparation of red yeast high-activity complex liposomes:

[0209] 100 mg of soybean lecithin was dissolved in 10 mL of chloroform to obtain a 10% mass concentration of lecithin chloroform solution, and the solvent was removed at 40°C using a rotary evaporator to form a lecithin film; 10 mg of red yeast high-activity complex was then added to the film, and ultrasonic treatment was performed for 15 minutes to form liposomes. Finally, the supernatant was removed by centrifugation (3000 rpm, 10 minutes) to remove the unencapsulated complex and excess phospholipids, and the collected liposomes were resuspended in an appropriate amount of PBS buffer for subsequent detection.

[0210] (2) Preparation of red yeast high-activity complex vesicles:

[0211] Polyvinyl alcohol (PVA) was dissolved in deionized water to obtain a 4% PVA aqueous solution, 10 mg of the prepared red yeast high-activity complex was added to the PVA solution, and stirred at 1000 r / min for 30 min; the mixture was converted into solid vesicles by spray drying (temperature controlled at 60-80°C), and large particles were sieved through a 60-mesh sieve to obtain a vesicle complex for subsequent experiments.

[0212] (3) Preparation of red yeast high-activity complex bile body:

[0213] Phospholipids and bile salts (taurocholic acid) were mixed in a mass ratio of 1:1 and dissolved in deionized water. Then, red yeast high-activity complex (50 mg) was gradually added to the mixture and mixed evenly under high shear stirring. Ultrasonic treatment was used for 20 minutes to promote the formation of bile bodies. Subsequently, centrifugation was performed at 12000 r / min for 30 minutes to remove unencapsulated components, and the supernatant was collected to obtain the bile body complex for subsequent analysis.

[0214] (4) Preparation of red yeast high activity composite microspheres:

[0215] Polylactic acid-glycolic acid copolymer (PLGA) was dissolved in an organic solvent, dichloromethane, to form a 10% PLGA solution. 50 mg of a red yeast high-activity complex was added to the PLGA solution and stirred at 1000 r / min for 30 min to form an emulsion. Deionized water was slowly added to the PLGA solution to obtain a precursor of emulsion microspheres. The dichloromethane was removed by freeze-drying to finally obtain microspheres. The microspheres were sieved with an 80 μm mesh and centrifuged at 10,000 r / min for 20 min to separate the microspheres. Finally, microspheres of appropriate particle size were collected for subsequent studies.

[0216] 2. Performance test of red yeast high-activity complex in other carrier forms

[0217] The red yeast highly active complex was prepared into various carrier forms, and their stability and release characteristics were evaluated respectively.

[0218] (1) Stability test

[0219] The stability test was carried out at 4°C, 25°C, and 50°C (dark conditions). The 7-day average retention rate of red yeast rhodotorula in different carriers is shown in Table 6:

[0220] Table 6 Average retention rate of red yeast rhodotorula in different carriers of F-EPA of Example 3 and Comparative Example 2 in darkness for 7 days (%)

[0221]

[0222] The red yeast high-activity composite corn oil nanoemulsion alginate gel (F-EPA) of Example 3 exhibits higher stability and can more effectively protect the red yeast erythrin in the red yeast high-activity composite and avoid degradation of the active ingredients.

[0223] (2) Release characteristics test

[0224] In addition, the release test results are shown in Table 7:

[0225] Table 7 Release rate of rhodozyma rhodozyma in different carriers of F-EPA of Example 3 and Comparative Example 4 after 2 hours of digestion in the small intestine (%)

[0226]

[0227] Although liposomes, vesicles, bile bodies, microspheres and microcapsules all successfully encapsulated the red yeast high-activity complex, the red yeast high-activity complex corn oil nanoemulsion alginate gel (F-EPA) of Example 3 was released more slowly, and under simulated physiological conditions (PBS buffer at pH 7.4), the red yeast erythrin in the encapsulated high-activity complex was able to safely reach the colon, showing the best effect.

[0228] Example 4: Preparation of soybean oil loaded with red yeast high activity complex (D-oil), red yeast high activity complex soybean oil nanoemulsion spray dried powder (D-EP), and red yeast high activity complex soybean oil nanoemulsion alginate gel (D-EPA)

[0229] According to the preparation method of Examples 1 to 3, the difference is that soybean oil is used instead of corn oil to prepare three carriers, namely soybean oil loaded with red yeast high activity complex (D-oil), red yeast high activity complex soybean oil nanoemulsion spray dried powder (D-EP), and red yeast high activity complex soybean oil nanoemulsion alginate gel (D-EPA).

[0230] The stability test was carried out at 4°C, 25°C and 50°C (dark conditions). The results are shown in Table 8:

[0231] Table 8 Average retention rate of red yeast rhodotorula in different carriers of Example 7 in the dark for 7 days (%)

[0232]

[0233] In addition, a release test was conducted, and after 2 hours of digestion in the small intestine, the release rates of red yeast rhodotorula in D-oil, D-EP, and D-EPA were 53.2%, 67.8%, and 73.5%, respectively, indicating that the carrier prepared with soybean oil can also be used as a potential colon delivery system to a certain extent.

[0234] Example 5: Preparation of sunflower oil loaded with red yeast high activity complex (K-oil), red yeast high activity complex sunflower oil nanoemulsion spray dried powder (K-EP), and red yeast high activity complex sunflower oil nanoemulsion alginate gel (K-EPA)

[0235] According to the preparation methods of Examples 1 to 3, the difference is that sunflower oil is used instead of corn oil to prepare three carriers, namely sunflower oil loaded with red yeast high activity complex (K-oil), red yeast high activity complex sunflower oil nanoemulsion spray-dried powder (K-EP), and red yeast high activity complex sunflower oil nanoemulsion alginate gel (K-EPA).

[0236] The stability test was carried out at 4°C, 25°C and 50°C (dark conditions). The results are shown in Table 9:

[0237] Table 9 Average retention rate of red yeast rhodotorula in different carriers of Example 8 in the dark for 7 days (%)

[0238]

[0239] In addition, release tests were conducted, and after 2 hours of digestion in the small intestine, the release rates of red yeast rhodotorula in K-oil, K-EP, and K-EPA were 51.3%, 62.7%, and 77.6%, respectively, indicating that the carrier prepared from sunflower oil can also be used as a potential colon delivery system to a certain extent.

[0240] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A method for preparing a red yeast high-activity complex carrier, characterized in that: The carrier is a vegetable oil loaded with a red yeast high-activity complex, comprising the following steps: (1) mixing red yeast red pigment and red yeast polysaccharide to obtain a red yeast high-activity complex; (2) dissolving the red yeast highly active complex obtained in step (1) in vegetable oil to obtain vegetable oil loaded with the red yeast highly active complex.

2. The preparation method according to claim 1, characterized in that: In step (1), red yeast rhodotorula rubrum and red yeast polysaccharide are mixed in a mass ratio of (8-12):

1.

3. The preparation method according to claim 1, characterized in that: In step (2), the vegetable oil includes but is not limited to corn oil, soybean oil, and sunflower oil.

4. The preparation method according to claim 1, characterized in that: In step (2), the mass ratio of the red yeast high-activity complex to the vegetable oil is (3-5):1000.

5. A method for preparing a red yeast high-activity complex carrier, characterized in that: The carrier is a red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder, comprising the following steps: (1) mixing red yeast red pigment and red yeast polysaccharide to obtain a red yeast high-activity complex; (2) dissolving the red yeast highly active complex obtained in step (1) in vegetable oil to obtain vegetable oil loaded with the red yeast highly active complex; (3) mixing the OSA starch solution with the vegetable oil loaded with the red yeast highly active complex, and homogenizing to obtain a red yeast highly active complex vegetable oil nanoemulsion; (4) spray-drying the red yeast high-activity complex vegetable oil nanoemulsion to obtain the red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder.

6. The preparation method according to claim 5, characterized in that: The OSA starch solution and the vegetable oil loaded with the red yeast high-activity complex are mixed in a mass ratio of 1:(8-10).

7. A method for preparing a red yeast high-activity composite carrier, characterized in that: The carrier is a red yeast high-activity complex plant oil nanoemulsion alginate gel, comprising the following steps: (1) mixing red yeast red pigment and red yeast polysaccharide to obtain a red yeast high-activity complex; (2) dissolving the red yeast highly active complex obtained in step (1) in vegetable oil to obtain vegetable oil loaded with the red yeast highly active complex; (3) mixing the OSA starch solution with the vegetable oil loaded with the red yeast highly active complex, and homogenizing to obtain a red yeast highly active complex vegetable oil nanoemulsion; (4) mixing the alginate solution with the red yeast high-activity complex vegetable oil nanoemulsion, and gelling to obtain the red yeast high-activity complex vegetable oil nanoemulsion alginate gel.

8. The preparation method according to claim 7, characterized in that: The alginate solution and the red yeast high-activity complex vegetable oil nanoemulsion are mixed in a mass ratio of (1-2):(1-2), homogenized, and stirred to form a gel block to obtain the red yeast high-activity complex vegetable oil nanoemulsion alginate gel.

9. Use of the vegetable oil loaded with red yeast high-activity complex prepared by the preparation method of any one of claims 1 to 4, the red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder prepared by the preparation method of claim 5 or 6, and the red yeast high-activity complex vegetable oil nanoemulsion alginate gel prepared by the preparation method of claim 7 or 8 in improving the stability of red yeast rhodotorula erythrin.

10. Use of the vegetable oil loaded with red yeast high-activity complex prepared by the preparation method of any one of claims 1 to 4, the red yeast high-activity complex vegetable oil nanoemulsion spray-dried powder prepared by the preparation method of claim 5 or 6, and the red yeast high-activity complex vegetable oil nanoemulsion alginate gel prepared by the preparation method of claim 7 or 8 in the preparation of products that promote gastrointestinal metabolic function.

Images