A method for preparing yeast microcapsules loaded with nosenoside

Abstract

This invention discloses a method for preparing yeast microcapsules loaded with hesperidin, belonging to the field of microcapsule technology. The invention includes the following steps: taking yeast powder and using yeast microcapsules obtained through alkali and acid treatment as the encapsulation wall material; adsorbing hesperidin using a metal polyphenol network self-assembled from epigallocatechin gallate and iron ions; and loading the hesperidin bound to the metal polyphenol network into the yeast microcapsules using a one-pot method. Microcapsulation of hesperidin not only preserves its antioxidant, anti-inflammatory, blood pressure-lowering, blood sugar-lowering, and memory-improving functional activities, but also provides a solution to the problem of hesperidin's low bioavailability due to its insolubility in water and its loss of activity due to oxidation upon heating or light exposure, thus increasing the stability of hesperidin.

Description

Technical Field

[0001] This invention belongs to the field of microcapsule technology, specifically relating to a method for preparing yeast microcapsules loaded with hesperidin. Background Technology

[0002] Nobiletin is a food-derived polymethoxyflavonoid extracted from citrus plants. It possesses various biological activities, including lowering blood pressure and blood sugar, antioxidation, anti-inflammation, improving memory, and penetrating the blood-brain barrier. Due to its broad therapeutic effects in various disease models, there is increasing interest in its application as a functional ingredient in drugs and foods. However, oral administration faces challenges such as hydrophobicity, instability in physiological media, and low gastrointestinal absorption, limiting its efficacy. Therefore, modifying nobiletin using physicochemical methods to obtain a nobiletin with high bioavailability is of significant practical importance for production.

[0003] Currently, many applications utilize yeast microencapsulation technology to encapsulate bioactive substances and deliver them to disease sites. Microencapsulation is a widely used technology in the pharmaceutical industry and is beginning to see widespread application in the food industry. Microcapsules encapsulate active substances within a tiny space. Encapsulation imparts a degree of stability to active compounds, and the wall material acts as a physical barrier against oxygen or other molecules, preventing harmful reactions and controlling the release of the core material. The stability and release properties of microcapsules are highly dependent on the composition of the wall material. Various carriers have been used, including cyclodextrin, maltodextrin, modified starch, gums, proteins, nanoparticles, micelles, and liposomes. However, these methods often suffer from problems such as chemical reagent residues and high reagent and equipment costs.

[0004] In recent years, yeast has been identified as a novel, safe, and edible microencapsulation biomaterial. Studies have shown that the presence of polysaccharides and proteins in yeast cell walls can promote their binding with various bioactive substances, such as curcumin, resveratrol, chlorogenic acid, berberine, and purslane seed oil, which have already been reported. The main component of the cell wall, polysaccharides, can protect the encapsulated substances from degradation in the acidic environment of the stomach, thereby promoting their retention and absorption in the intestines.

[0005] However, there are no reports on the encapsulation of nodosumin in yeast microcapsules, either domestically or internationally. Therefore, given the increasing public concern about food and drug safety, there is a need for a novel, green, and efficient method for encapsulating nodosumin in yeast microcapsules. Summary of the Invention

[0006] Technical issues

[0007] The encapsulation efficiency of the core material refers to the ratio of the content of the encapsulated core material in a microcapsule product to the total content of the core material in the microcapsule product. It is an indicator for measuring the encapsulation effect. Existing technologies use yeast to encapsulate active substances such as chlorogenic acid. While the method is feasible, the encapsulation efficiency is low, around 18.9%, failing to achieve the goal of highly efficient encapsulation of active substances. Therefore, this invention aims to overcome the drawback of low encapsulation efficiency.

[0008] Technical content

[0009] This invention provides a yeast microcapsule loaded with hesperidin and its preparation method. The microcapsule uses a metal polyphenol network (MSN, which in this invention is a metal polyphenol network formed by EGCG and FeCl3·6H2O) with good adsorption properties as an intermediate carrier to adsorb hesperidin as the core material, and yeast cells as the encapsulation wall material. The yeast microcapsule product loaded with hesperidin is obtained by constant temperature shaking in a water bath, centrifugation, and freeze drying.

[0010] This invention provides a method for preparing yeast microcapsules loaded with hesperidin, comprising the following steps:

[0011] S1: Mix yeast powder and alkaline solution, shake, centrifuge, take the precipitate, wash with water, then put the precipitate into water, add hydrochloric acid to adjust the pH of the solution to 4-5, then incubate, centrifuge, take the precipitate again, wash with water, then wash with organic solvent, and then dry the precipitate to obtain yeast microcapsule powder.

[0012] S2: Dissolve hesperidin in Tris buffer of ethanol to obtain hesperidin solution, slowly add EGCG solution to hesperidin solution, mix evenly, then add FeCl3·6H2O solution, mix evenly to obtain NOB@MSN nano solution;

[0013] S3: Add the yeast microcapsule powder obtained in step S1 to the NOB@MSN nano solution obtained in step S2 and mix well. Shake, centrifuge, wash, and freeze dry to obtain yeast microcapsules loaded with hesperidin.

[0014] Furthermore, in step S1, the alkaline solution is one or more of NaOH, KOH, and Ca(OH)2 aqueous solution.

[0015] Specifically, the alkaline solution in step S1 can be an aqueous solution of NaOH.

[0016] Furthermore, the concentration of the alkaline solution in step S1 is 0.5–2 mol / L.

[0017] Furthermore, in step S1, the mass ratio of yeast powder to alkaline solution is 1:5 to 15.

[0018] Furthermore, in step S1, oscillation refers to oscillation in a water bath at 70–80°C for 0.5–1 hour.

[0019] Furthermore, in step S1, the mass ratio of precipitate to water is 1:5 to 15.

[0020] Furthermore, the concentration of hydrochloric acid in step S1 is 1–6 mol / L.

[0021] Furthermore, in step S1, incubation refers to incubation in a water bath at 50–55°C for 0.5–1 hour.

[0022] Furthermore, in step S1, the organic solvent is one or more of isopropanol, acetone, and ethanol.

[0023] Furthermore, in step S1, the amount of water used to wash the precipitate is 1 to 10 times the mass of the precipitate.

[0024] Furthermore, in step S1, the amount of organic solvent used to wash the precipitate is 1 to 10 times the mass of the precipitate.

[0025] Furthermore, in step S2, the concentration of ethanol in the Tris buffer is 40-60%, the concentration of Tris is 5-15 mM, and the pH is 7-7.5.

[0026] Furthermore, the concentration of the noriheptacortine solution in step S2 is 0.5–2.5 mg / mL.

[0027] Preferably, the concentration of the noriheptacortine solution in step S2 is 1.25 mg / mL.

[0028] Furthermore, in step S2, the concentration of the EGCG solution is 20–80 mM.

[0029] Furthermore, in step S2, the concentration of the FeCl3·6H2O solution is 20–80 mM.

[0030] Preferably, the concentration of the EGCG solution in step S2 is 40 mM.

[0031] Preferably, the concentration of the FeCl3·6H2O solution in step S2 is 40 mM.

[0032] Furthermore, in step S2, the volume ratio of the hesperidin solution to the EGCG solution is 10 mL: 100–300 μL.

[0033] Furthermore, in step S2, the volume ratio of the hesperidin solution and the FeCl3·6H2O solution is 10 mL: 100–300 μL.

[0034] Furthermore, in step S3, the mass-to-volume ratio of yeast microcapsule powder to NOB@MSN nanosolution is 20–70 mg: 10 mL.

[0035] Furthermore, in step S3, oscillation refers to oscillation in a constant temperature water bath at 25–50°C for 1–24 hours.

[0036] This invention provides yeast microcapsules loaded with hesperidin prepared by the above method.

[0037] The yeast microcapsules loaded with hesperidin provided by this invention have applications in the food and pharmaceutical preparation fields.

[0038] Furthermore, the drug preparation refers to the preparation of a drug containing nodosumetin.

[0039] Beneficial effects

[0040] 1. The yeast microcapsules loaded with hesperidin obtained by the method described above have a high encapsulation efficiency, which improves the water solubility of hesperidin. These microcapsules are safe, easily degradable, and possess antioxidant, anti-inflammatory, blood pressure-lowering, blood sugar-lowering, and memory-improving functional activities. They can target disease sites and control release, making them suitable for mass production.

[0041] 2. The yeast microcapsules loaded with hesperidin obtained by this invention can exert antioxidant, anti-inflammatory, blood pressure lowering, blood sugar lowering, and memory-improving functional activities while improving the encapsulation rate of active substances; and avoids the food and drug safety problems caused by chemical reagent residues in traditional encapsulation methods. It is a novel, green, and safe method for encapsulating active substances.

[0042] 3. This invention is based on repeated experiments. The concentration and ratio of hesperidin to MSN, the reaction time and temperature of NOB@MSN with yeast microcapsules, and the order of each step all directly affect the encapsulation efficiency and functional activity of the resulting yeast microcapsules loaded with hesperidin. Attached Figure Description

[0043] Figure 1 This is the standard curve of the concentration of noriheptacortin in this invention.

[0044] Figure 2 This invention relates to the encapsulation rate and loading rate of yeast microcapsules loaded with 1.25 mg / ml hesperidin, obtained by reaction with different concentrations of MSN without or with no added hesperidin.

[0045] Figure 3 This invention relates to the encapsulation rate and loading rate of yeast microcapsules loaded with hesperidin obtained by reacting hesperidin with metal polyphenols at a fixed ratio and different concentrations of hesperidin.

[0046] Figure 4These are transmission electron micrographs of yeast powder, yeast microcapsules, yeast microcapsules loaded with hesperidin obtained by MSN treatment, and yeast microcapsules loaded with hesperidin obtained without MSN treatment, all obtained in this invention.

[0047] Figure 5 This is a scanning electron microscope image of yeast microcapsules loaded with hesperidin obtained by processing yeast powder, yeast microcapsules, and MSN in this invention.

[0048] Figure 6 This is an analysis of the energy dispersive spectroscopy (EDS) results of the yeast microcapsules in this invention.

[0049] Figure 7 This is an analysis of the energy dispersive spectroscopy (EDS) results of yeast microcapsules loaded with hesperidin obtained by MSN processing in this invention.

[0050] Figure 8 This paper analyzes the antioxidant effects of yeast microcapsules loaded with hesperidin obtained by MSN treatment in this invention, which scavenges hydroxyl radicals. Detailed Implementation

[0051] The invention will be further illustrated below through specific implementation examples.

[0052] Source of raw materials

[0053] The tangeretin was purchased from Shanghai Yuanye Biotechnology Co., Ltd.; EGCG was purchased from Shanghai Maclean Biochemical Technology Co., Ltd.; FeCl3 was purchased from Tianjin Damao Chemical Reagent Factory; and yeast powder was purchased from Angel Yeast Co., Ltd.

[0054] Testing process

[0055] The absorbance of hesperidin at 340 nm was measured using a UV spectrophotometer at concentration gradients ranging from 0.0025 mg / mL to 0.1 mg / mL, and a standard curve for hesperidin concentration was established. The encapsulation efficiency and loading rate of the yeast microcapsules loaded with hesperidin were calculated using the following formula:

[0056] Encapsulation efficiency = (Total mass of hesperidin before encapsulation - Mass of hesperidin in the supernatant after encapsulation) / Total mass of hesperidin before encapsulation × 100%

[0057] Loading rate = (Total mass of hesperidin before encapsulation - Mass of hesperidin in the supernatant after encapsulation) / Total mass of yeast microcapsule powder × 100%

[0058] The total mass of hesperidin before encapsulation was calculated by substituting the absorbance value of the solution measured at 340 nm using a spectrophotometer into the standard curve. The mass of hesperidin in the supernatant after encapsulation was calculated by substituting the absorbance value of the supernatant obtained by centrifuging the yeast microcapsules and NOB@MSN and removing the metal polyphenol network by EDTA into the standard curve.

[0059] Example 1

[0060] S1: Take yeast powder, add 10 times its mass of 1 mol / L NaOH solution, shake in an 80℃ water bath for 1 hour, centrifuge at 2000g for 10 minutes, take the precipitate and wash it twice with deionized water; then suspend the precipitate in 10 times its mass of deionized water, add 3 mol / L hydrochloric acid to adjust the pH of the solution to 4.5, incubate in a 55℃ water bath for 1 hour, centrifuge again, take the precipitate, wash the precipitate 3 times with 10 times its mass of deionized water, wash it 4 times with 2 times its mass of isopropanol, wash it 2 times with 2 times its mass of acetone, and finally dry the precipitate at room temperature to obtain yeast microcapsule powder.

[0061] S2: Dissolve hesperidin in 10 mL of 60% ethanol Tris buffer (10 mM, pH 7.5) to form a 1.25 mg / mL hesperidin solution. Then, slowly add 200 μL of EGCG solution at different concentrations (0, 10, 20, 40, 60, 80, 100, 200, 400 mM) to each group and mix thoroughly to obtain a mixture. Then, add 200 μL of FeCl3·6H2O solution at the same concentration as EGCG (0, 10, 20, 40, 60, 80, 100, 200, 400 mM) to the mixture and shake vigorously for 60 s under ultrasonication to obtain a NOB@MSN nanosolution.

[0062] S3: Take 50 mg of yeast microcapsule powder obtained in step S1 and add it to the NOB@MSN nano solution obtained in step S2 and mix well. After shaking and embedding in a constant temperature water bath at 30℃ for 12 h, centrifuge, wash, and freeze dry to obtain yeast microcapsules loaded with hesperidin.

[0063] The encapsulation efficiency and loading rate of hesperidin loaded onto yeast microcapsules were calculated, and the results are as follows: Figure 2 And as shown in Table 1 below:

[0064] Table 1. Effects of different MSN concentrations on the encapsulation efficiency and loading rate of norepinephrine loaded onto yeast microcapsules.

[0065] MSN / mM 0 10 20 40 60 80 100 200 400 Encapsulation rate / % 7.37 25.96 28.63 38.24 32.99 28.57 21.99 13.90 5.37 Load rate / % 1.84 6.50 7.16 9.56 8.25 7.14 5.44 3.48 1.34

[0066] The results showed that the encapsulation efficiency and loading rate of norepinephrine were highest when the concentration of EGCG and FeCl3·6H2O was 40 mM. This indicates that under this condition, the nanoparticles formed by norepinephrine and MSN (EGCG and FeCl3·6H2O) can ensure that the maximum amount of norepinephrine is encapsulated in the yeast microcapsules. Therefore, the final molar concentration ratio of norepinephrine to MSN under this condition (calculated to be 1:2.69) was selected as the optimal encapsulation ratio.

[0067] Example 2

[0068] S1: Noriheptacorlina was dissolved in 10 mL of 60% ethanol in Tris buffer (10 mM, pH 7.5) to form noriheptacorlina solutions of different concentrations (0.5, 0.625, 1.25, 2.5, 5, 10 mg / mL). Then, EGCG solution (40 mM, 200 μL) was slowly added to each group, and the mixture was homogenized to obtain a final solution. Next, FeCl3·6H2O solution (40 mM, 200 μL) was added to the mixture, and the mixture was vigorously shaken under ultrasonication for 60 s to obtain the NOB@MSN nanosolution. The final concentration ratio of noriheptacorlina to EGCG or FeCl3·6H2O was 1:2.69. Figure 2 The proportion at the highest encapsulation rate.

[0069] S2: Take 50 mg of yeast microcapsule powder obtained in step S1 of Example 1, add it to the NOB@MSN nano solution obtained in step S1 and mix well. After shaking and embedding in a constant temperature water bath at 30°C for 12 h, centrifuge, wash, and freeze dry to obtain yeast microcapsules loaded with hesperidin.

[0070] The encapsulation efficiency and loading rate of hesperidin loaded onto yeast microcapsules were calculated, and the results are as follows: Figure 3 And as shown in Table 2 below:

[0071] Table 2. Effects of different concentrations of norepinephrine solution on the encapsulation efficiency and loading rate of norepinephrine loaded onto yeast microcapsules.

[0072]

[0073] The results showed that as the concentration of norepinephrine increased, the encapsulation efficiency of norepinephrine gradually decreased, and the loading first decreased and then increased. Therefore, considering all factors, the optimal concentration of norepinephrine solution was 1.25 mg / mL. Note: The data in column 5 of Table 1 in Example 1 and the data in column 4 of Table 2 in Example 2 are both optimal results, and the differences in data are only due to experimental error.

[0074] Example 3

[0075] A method for preparing yeast microcapsules loaded with noriheptacorlina includes the following steps:

[0076] S1: Take yeast powder, add 10 times its mass of 1 mol / L NaOH solution, shake in an 80℃ water bath for 1 hour, centrifuge at 2000g for 10 minutes, take the precipitate and wash it twice with deionized water; then suspend the precipitate in 10 times its mass of deionized water, add 3 mol / L hydrochloric acid to adjust the pH of the solution to 4.5, incubate in a 55℃ water bath for 1 hour, centrifuge again, take the precipitate, wash the precipitate 3 times with 3 times its mass of deionized water, wash it 4 times with 2 times its mass of isopropanol, wash it 2 times with 2 times its mass of acetone, and finally dry the precipitate at room temperature to obtain yeast microcapsule powder.

[0077] S2: Dissolve tangeretin in 10 mL of 60% ethanol Tris buffer (10 mM, pH 7.5) to form a 1.25 mg / mL tangeretin solution. Then slowly add EGCG solution (200 μL, 40 mM) and mix evenly to obtain a mixture. Then, add FeCl3·6H2O solution (200 μL, 40 mM) to the mixture and shake under ultrasonication for 60 s to obtain NOB@MSN nano solution.

[0078] S3: Take 50 mg of yeast microcapsule powder obtained in step S1 and add it to the NOB@MSN nano solution obtained in step S2 and mix well. After shaking and embedding in a constant temperature water bath at 30℃ for 12 h, centrifuge, wash, and freeze dry to obtain yeast microcapsules loaded with noriheptacorlin.

[0079] The encapsulation rate of the yeast microcapsules loaded with hesperidin obtained in this embodiment was 38.24%, and the loading rate was 9.56%. Transmission electron microscopy, scanning electron microscopy, energy dispersive spectroscopy and antioxidant activity were also performed.

[0080] The transmission electron microscopy (TEM) images are attached. Figure 4 As shown in the figure, Yeast represents yeast powder, YM represents yeast microcapsule powder, and NOB@MSN@YM and NOB@YM represent yeast microcapsules loaded with hesperidin with or without the addition of metal polyphenols. It can be seen that compared to Yeast, most substances in the cytoplasm of YM obtained after alkali and acid treatment dissolved, providing ample space for loading the active substance. In the transmission electron microscopy image of NOB@YM, very little hesperidin is loaded; most of the hesperidin aggregates and adsorbs on the periphery of the yeast microcapsules. In contrast, the NOB@MSN@YM group with the added metal polyphenol network shows a significant loading of hesperidin in the cytoplasm of the yeast microcapsules, with no aggregated hesperidin observed on the periphery. Therefore, it can be concluded that metal polyphenols enhance the loading of the active substance hesperidin onto yeast microcapsules.

[0081] The scanning electron microscope (SEM) images are attached. Figure 5As shown, compared to yeast, the surface of YM obtained after alkali and acid treatment is wrinkled, while the surface of the yeast microcapsules in the NOB@MSN@YM group loaded with norihesperidin is more plump, further verifying the successful loading of norihesperidin. The energy dispersive spectroscopy (EDS) results are attached. Figure 6 and 7 As shown, the attached Figure 6 The test was conducted on the yeast microcapsules prepared in step S1 of Example 3, with attached... Figure 7 The test was conducted on the yeast microcapsules loaded with hesperidin prepared in step S3 of Example 3. It can be seen that there is iron in the metal polyphenols, and the proportion of O element increases due to the loading of hesperidin.

[0082] The antioxidant test results are attached. Figure 8 As shown, Control represents the electron paramagnetic resonance (EPR) spectrum of hydroxyl radicals in aqueous solution, while YM and NOB@MSN@YM represent the EPR spectra of hydroxyl radicals under the intervention of yeast microcapsules and yeast microcapsules loaded with hesperidin, respectively. It can be seen that yeast microcapsules have a certain ability to scavenge hydroxyl radicals, and yeast microcapsules loaded with hesperidin exhibit a more significant effect in scavenging hydroxyl radicals. This indicates that yeast microcapsules loaded with hesperidin possess significant antioxidant capacity.

[0083] Comparative Example 1

[0084] A method for preparing yeast microcapsules loaded with noriheptacorlina includes the following steps:

[0085] S1: Dissolve tangeretin in 10 mL of 60% ethanol in Tris buffer (10 mM, pH 7.5) to form a 1.25 mg / mL tangeretin solution.

[0086] S3: Take 50 mg of yeast microcapsule powder obtained in step S1 of Example 1, add it to the norihesperidin solution obtained in step S1 and mix well. After shaking and embedding in a constant temperature water bath at 37°C for 6 hours, centrifuge, wash, and freeze dry to obtain the yeast microcapsules loaded with norihesperidin.

[0087] The encapsulation rate and loading rate of the yeast microcapsules loaded with nobiletin obtained by the above method were extremely low, calculated to be 7.3% and 1.8%, respectively.

[0088] Comparative Example 2

[0089] A method for preparing yeast microcapsules loaded with noriheptacorlina includes the following steps:

[0090] S1: Dissolve hesperidin in 10 mL of 60% ethanol in Tris buffer (10 mM, pH 7.5) to form a 10 mg / mL hesperidin solution. Slowly add EGCG solution (200 μL, 320 mM) to the hesperidin solution and mix thoroughly. Then, add FeCl3·6H2O solution (200 μL, 160 mM) and shake under ultrasonication for 60 s to obtain NOB@MSN nanosolution.

[0091] S3: Take 50 mg of yeast microcapsule powder obtained in step S1 of Example 1, add it to the NOB@MSN nano solution obtained in step S1 and mix well. After shaking and embedding in a constant temperature water bath at 50°C for 24 h, centrifuge, wash, and freeze dry to obtain the yeast microcapsules loaded with hesperidin.

[0092] The encapsulation efficiency of the yeast microcapsules loaded with nobiletin obtained by the above method was extremely low, calculated to be 6.5%, with a loading rate of 13%.

[0093] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for preparing yeast microcapsules loaded with noriheptacorlina, characterized in that, Includes the following steps: S1: Mix yeast powder and alkaline solution, shake, centrifuge, take the precipitate, wash with water, then put the precipitate into water, add hydrochloric acid to adjust the solution pH to 4-5, then incubate, centrifuge, take the precipitate again, wash with water, then wash with organic solvent, and then dry the precipitate to obtain yeast microcapsule powder. S2: Dissolve hesperidin in Tris buffer of ethanol to obtain hesperidin solution. Slowly add EGCG solution to the hesperidin solution and mix evenly. Then, add FeCl3·6H2O solution and shake vigorously for 60 s under ultrasonication to obtain NOB@MSN nano solution. The concentration of hesperidin solution is 0.5~2.5 mg / mL; the concentration of EGCG solution is 20~80 mM; the concentration of FeCl3·6H2O solution is 20~80 mM. The volume ratio of hesperidin solution to EGCG solution was 10 mL: 100~300 μL; the volume ratio of hesperidin solution to FeCl3·6H2O solution was 10 mL: 100~300 μL. S3: Add the yeast microcapsule powder obtained in step S1 to the NOB@MSN nanosolution obtained in step S2 and mix well. Shake, centrifuge, wash, and freeze dry to obtain yeast microcapsules loaded with hesperidin. The mass-volume ratio of yeast microcapsule powder to NOB@MSN nanosolution is 20~70 mg:10 mL. Shaking refers to shaking in a constant temperature water bath at 25~30℃ for 1~12 h.

2. The method according to claim 1, characterized in that, The alkaline solution in step S1 is one or more of NaOH, KOH and Ca(OH)2 aqueous solution; the concentration of the alkaline solution in step S1 is 0.5~2 mol / L.

3. The method according to claim 1, characterized in that, In step S1, the mass ratio of yeast powder to alkaline solution is 1:5~15.

4. The method according to claim 1, characterized in that, In step S1, the organic solvent is one or more of isopropanol, acetone, and ethanol; the amount of water used to wash the precipitate in step S1 is 1 to 10 times the mass of the precipitate; and the amount of organic solvent used to wash the precipitate in step S1 is 1 to 10 times the mass of the precipitate.

5. The method according to claim 1, characterized in that, In step S2, the concentration of ethanol in the Tris buffer is 40-60%, the concentration of Tris is 5-15 mM, and the pH is 7-7.

5.

6. Yeast microcapsules loaded with hesperidin prepared by the method according to any one of claims 1 to 5.

7. The use of the yeast microcapsules loaded with hesperidin as described in claim 6 in the preparation of food or medicine.

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