A method for improving the reconstitution performance of ginsenoside microcapsules and the bioavailability of ginsenoside Rg3 and CK

Abstract

The application discloses a method for improving the reconstitution performance of ginsenoside microcapsules and the bioavailability of ginsenoside Rg3 and CK, and belongs to the field of functional food. In the method, the rare ginsenoside Rg3 and CK are extracted by acid hydrolysis, and then ginsenoside extract containing the rare ginsenoside Rg3 and CK and edible oil are used as an oil phase, whey protein isolate and Tween 80 are used as emulsifiers, and ginsenoside emulsion is prepared. On the basis of the emulsion, ginsenoside microcapsules are prepared by spray drying with maltodextrin and inulin / lactose as composite wall materials. The ginsenoside microcapsules are light yellow solid powder and have good reconstitution performance. The reconstituted emulsion has high ginsenoside Rg3 and CK embedding rate, strong stability and high ginsenoside Rg3 and CK bioavailability. In addition, the method is simple in operation, safe in material, conforms to actual industrial production and has good market prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to a method for improving the reconstitution performance of ginsenoside microcapsules and the bioavailability of ginsenoside Rg3 and CK, belonging to the field of functional foods. BACKGROUND

[0002] Ginsenosides are the main active substances of ginseng for physiological functions and important indicators for evaluating the quality of ginseng. Ginsenosides are triterpenoids composed of hydrophobic aglycone (lower polarity) and hydrophilic sugar (higher polarity), which have multiple pharmacological effects such as anticancer, anti-fatigue, anti-inflammatory, anti-diabetic, immune enhancement, memory improvement, and cardiovascular protection. According to the difference in content, ginsenosides are divided into major ginsenosides and rare ginsenosides. Major ginsenosides contain more sugar groups, have stronger polarity and better water solubility, but have limited functional activity and low oral bioavailability. Rare ginsenosides have less sugar groups, weaker polarity and strong functional activity, but their content is extremely low. Although rare ginsenosides have strong drug activity and good cell membrane permeability, their poor water solubility limits their bioavailability and affects their physiological activity in vivo.

[0003] For natural active substances with poor water solubility, emulsion delivery system is the most commonly used method to improve their bioavailability. However, with the extension of storage time, the stability of emulsion decreases, and the entrapment rate of the entrapped active substances also decreases, affecting the delivery of nutrients, and emulsion is also easily contaminated by microorganisms. To solve the problems of emulsion, microcapsule technology is an effective method to improve the shortcomings of emulsion delivery system. By adding suitable wall materials to emulsion and using spray drying to prepare microcapsules, the shelf life can be effectively prolonged, and stable performance can be maintained. The type and amount of wall materials play an important role in the physicochemical properties, shelf life and functional properties of microcapsules, and spray drying has higher requirements for wall materials, such as low viscosity, good water solubility, and good film-forming property.

[0004] Maltodextrin is a food raw material produced by starch hydrolysis, which is commonly used as a wall material in microcapsule preparation. However, maltodextrin also has the disadvantage of poor emulsifying ability, which leads to poor stability of microcapsule reconstitution emulsion, and it is difficult to reproduce the internal structure of emulsion system, which is not conducive to the delivery of nutrients.

[0005] Redissolution properties include solubility, wettability, and dispersibility. Solubility is the ability of a powder to dissolve in a solution, wettability is the ability of a powder to absorb water and rehydrate without stirring, and dispersibility is the ability to disperse in water without agglomerating. The greater the solubility, the better the wettability (the shorter the wetting time), and the stronger the dispersibility, the better the redissolution properties of the microcapsules. Conventionally prepared microcapsules, while emphasizing their good solubility or water solubility, solubility is only one of the redissolution indicators and does not necessarily indicate good redissolution properties. Summary of the Invention

[0006] The present invention is based on preliminary experimental results: ginsenoside microcapsules prepared with maltodextrin alone as the wall material have poor resolubility, low embedding rates of ginsenosides Rg3 and CK in the re-dissolved emulsion, and poor stability.

[0007] Based on this, the present invention uses maltodextrin and inulin / lactose as composite wall materials, which can significantly improve the resolubility performance of ginsenoside microcapsules, increase the embedding rate of ginsenoside Rg3 and CK in the resolubilized emulsion, enhance the stability of the resolubilized emulsion, and thus improve the bioaccessibility of ginsenoside Rg3 and CK, which is also expected to further improve the in vivo bioavailability of ginsenosides.

[0008] More specifically, the present invention first uses acid hydrolysis to extract rare ginsenosides Rg3 and CK. The resulting saponin extract containing rare ginsenosides Rg3 and CK and edible oil are then used as the oil phase, with whey protein isolate (WPI) and Tween 80 as emulsifiers to prepare a ginsenoside emulsion. Based on the emulsion, maltodextrin and inulin / lactose are used as composite wall materials to prepare ginsenoside microcapsules by spray drying. The ginsenoside microcapsules prepared by the present invention are light yellow solid powders with good re-dissolution properties. The re-dissolved emulsion has a high ginsenoside Rg3 and CK embedding rate, strong stability, and high ginsenoside Rg3 and CK bioaccessibility. In addition, the method of the present invention is simple to operate, uses safe materials, is in line with actual industrial production, and has good market prospects.

[0009] The first object of the present invention is to provide a method for improving the resolubility of ginsenoside microcapsules and the bioaccessibility of ginsenosides Rg3 and CK, comprising the following steps:

[0010] The wall material and the emulsion containing rare ginsenosides Rg3 and CK were mixed uniformly and then spray-dried to obtain ginsenoside microcapsules;

[0011] The wall material is one of a mixture of maltodextrin and inulin and a mixture of maltodextrin and lactose.

[0012] In one embodiment of the present invention, the mass ratio of maltodextrin to inulin in the wall material is 1:2-4; the mass ratio of maltodextrin to lactose is 1:2-4.

[0013] In one embodiment of the present invention, the DE of the maltodextrin in the wall material is 15-20.

[0014] In one embodiment of the present invention, the usage ratio of the wall material and the emulsion containing rare ginsenosides Rg3 and CK is 15-30 g:100 mL.

[0015] In one embodiment of the present invention, the uniform mixing is performed by stirring at 200-400 rpm for 2-4 hours.

[0016] In one embodiment of the present invention, the process parameters of the spray drying are: inlet temperature of 150-190°C, outlet temperature of 80-100°C, air injection rate of 10-30m 3 / h, and the injection rate was 2-10mL / min.

[0017] In one embodiment of the present invention, the method for preparing the emulsion containing rare ginsenosides Rg3 and CK comprises the following steps:

[0018] (1) Water phase:

[0019] Mix WPI, Tween 80 and water to obtain an aqueous phase;

[0020] (2) Oil phase:

[0021] The saponin extract containing rare ginsenosides Rg3 and CK and edible oil are mixed uniformly to obtain an oil phase;

[0022] (3) Emulsion:

[0023] The aqueous phase and the oil phase are mixed and shear-dispersed to obtain a coarse emulsion; the coarse emulsion is then homogenized by microfluidization to obtain an emulsion containing rare ginsenosides Rg3 and CK;

[0024] The preparation of the saponin extract containing rare ginsenosides Rg3 and CK comprises the following steps:

[0025] After mixing ginseng slices with a citric acid solution with a pH of 1.0-3.0 or ultrapure water (pH of 6.12), the mixture is heated at 50-130°C for 1-5 hours to obtain a saponin extract containing rare ginsenosides Rg3 and CK; wherein the ratio of the ginseng slices to the citric acid solution is 1 g: 5-10 mL; the optimal extraction conditions for the method of acid hydrolysis and heating to extract rare ginsenosides Rg3 and CK are: pH 2.0, heating at 110°C for 3-5 hours, or pH 6.12, heating at 110°C for 3 hours;

[0026] The WPI, Tween 80 and water are used in a ratio of 0.5 g: 0.5-0.7 g: 50 mL;

[0027] The aqueous phase is prepared by dissolving WPI in water, heating at 80-90°C for 15-20 minutes, immediately cooling to room temperature in an ice-water bath, adding Tween 80, stirring at 200-400 rpm for 2-4 hours, and finally standing at 4°C for 10-12 hours until completely hydrated to obtain an aqueous phase;

[0028] The volume ratio of saponin extract containing rare ginsenosides Rg3 and CK to edible oil is 4:1;

[0029] Edible oils include one or more of rapeseed oil, soybean oil, and peanut oil;

[0030] The mixing was carried out by stirring at 200-400 rpm for 10-12 h;

[0031] The volume ratio of water phase to oil phase is 1:1;

[0032] Shear dispersion is performed at 15000-18000 rpm for 4-6 minutes;

[0033] Microfluidization homogenization is a 10-15 min circulating homogenization process at 600-800 bar, and the applicable sample volume is 150-200 mL.

[0034] The second object of the present invention is the ginsenoside microcapsules prepared by the method of the present invention.

[0035] The third object of the present invention is to provide a ginsenoside microcapsule reconstituted emulsion, wherein the emulsion is obtained by adding ginsenoside microcapsules to water and mixing uniformly;

[0036] The mass ratio of ginsenoside microcapsules to water is 15-30 g:100 mL;

[0037] The uniform mixing is carried out by stirring at 200-400 rpm for 2-4 hours.

[0038] The fourth object of the present invention is to use the ginsenoside microcapsules and ginsenoside microcapsule re-dissolved emulsion prepared by the present invention in the field of functional foods.

[0039] In one embodiment of the present invention, the application is to re-dissolve ginsenoside microcapsules to form an emulsion, and use it as a raw material in the mixing stage of beverage production, with an amount of 30-60% (mass percentage), which can enhance the immune function of the beverage.

[0040] In one embodiment of the present invention, the application is to redissolve ginsenoside microcapsules to form an emulsion, and use it as a raw material in the mixing stage of emulsion cosmetics production, with an amount of 1-10% (mass percentage), which can enhance the whitening and skin care effects of cosmetics.

[0041] The effects that can be achieved by the present invention are:

[0042] (1) Maltodextrin is a nutritious polysaccharide produced by mild hydrolysis of starch. It has excellent physical and chemical properties, such as good solubility, thickening, adhesion, and film-forming properties. It also has the functions of inhibiting browning reaction, reducing the sweetness of the system, preventing product agglomeration, and increasing product dispersibility. It has physiological functions such as lowering blood sugar and blood lipids, improving intestinal health, and promoting calcium absorption and utilization. Inulin is a natural prebiotic polysaccharide that exhibits excellent food processing properties, such as good solubility and water retention. It can be used as an emulsifier stabilizer and has the functions of improving intestinal microecology, regulating blood sugar levels, promoting mineral absorption, losing weight, and lowering blood lipids. Lactose is a common disaccharide that can be used as a filler, flow aid, disintegrant, lubricant, and adhesive. It can provide energy to the human body, promote the production of certain lactic acid bacteria and bifidobacteria in the human intestine, and inhibit the growth of spoilage bacteria.

[0043] (2) The ginsenoside microcapsules prepared by the present invention have good re-dissolution performance, and the re-dissolved emulsion thereof has a high ginsenoside Rg3 and CK embedding rate, strong stability, high ginsenoside Rg3 and CK bioaccessibility, and a long service life and practical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 These are the test results of the yields of rare ginsenosides Rg3 and CK in the saponin extracts obtained in Example 1 when the citric acid solution has a pH of 1.0, 2.0, 3.0 and 6.12 (ultrapure water) and the reaction temperature is 50, 70, 90, 110 and 130°C and heated for 1, 3 and 5 hours, respectively; wherein A is the yield of rare ginsenoside Rg3 after heating for 1 hour; B is the yield of rare ginsenoside Rg3 after heating for 3 hours; C is the yield of rare ginsenoside Rg3 after heating for 5 hours; D is the yield of rare ginsenoside CK after heating for 1 hour; E is the yield of rare ginsenoside CK after heating for 3 hours; and F is the yield of rare ginsenoside CK after heating for 5 hours.

[0045] Figure 2The wetting process of ginsenoside microcapsules prepared in Examples 3 and 4 and Comparative Example 1 is shown; wherein, A is the situation when the microcapsules prepared with maltodextrin: inulin = 4:0 as the wall material are wetted for 2s; B is the situation when the microcapsules prepared with maltodextrin: inulin = 1:3 as the wall material are wetted for 2s; C is the situation when the microcapsules prepared with maltodextrin: lactose = 1:3 as the wall material are wetted for 2s; D is the situation when the microcapsules prepared with maltodextrin: inulin = 4:0 as the wall material are wetted for 30s; E is the situation when the microcapsules prepared with maltodextrin: inulin = 1:3 as the wall material are wetted for 30s; F is the situation when the microcapsules prepared with maltodextrin: lactose = 1:3 as the wall material are wetted for 30s.

[0046] Figure 3 The release levels of free fatty acids in Example 6 at 0, 20, 40, 60, 80, 100, and 120 minutes in the intestinal digestion stage.

[0047] Figure 4 These are the test results of the bioaccessibility of ginsenosides Rg3 and CK in the products of the intestinal digestion stage of Example 6. DETAILED DESCRIPTION

[0048] The following describes preferred embodiments of the present invention. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention.

[0049] Test method:

[0050] ①Test of ginsenoside Rg3 and CK embedding rate

[0051] 2 mL of the emulsion sample was mixed with 8 mL of methanol and sonicated for 30 minutes. The mixture was then centrifuged at 10,000 rpm at 4°C for 30 minutes. The supernatant was filtered through a 0.45 μm organic membrane to remove impurities. The concentrations of ginsenosides Rg3 (C1) and CK (C2) in the filtrate were determined using high-performance liquid chromatography (elution program shown in Table 1). Each sample was run in triplicate.

[0052] Table 1 HPLC elution program

[0053] Time (min) Acetonitrile (%) Ultra-pure water (%) 0 22.0 78.0 25.00 22.0 78.0 35.00 32.5 67.5 65.00 34.0 66.0 66.00 42.0 58.0 114.00 98.0 2.0 119.00 98.0 2.0 120.00 22.0 78.0 130.00 22.0 78.0

[0054] The emulsion sample was centrifuged at 35,000 g for 30 minutes at 4°C to separate into three layers: an upper oil phase, a middle aqueous phase, and a lower impurity phase. The intermediate aqueous phase was filtered through a 0.45 μm membrane to remove impurities. The concentrations of ginsenosides Rg3 (C1′) and CK (C2′) in the filtrate were determined by high-performance liquid chromatography. Each sample was run in triplicate.

[0055] The embedding rates of ginsenoside Rg3 and CK in the emulsion samples were calculated by formulas (1) and (2), respectively:

[0056]

[0057]

[0058] Among them, EE1 is the embedding rate of ginsenoside Rg3 (%), EE2 is the embedding rate of ginsenoside CK (%), C1 is the concentration of ginsenoside Rg3 in methanol (ppm), C2 is the concentration of ginsenoside CK in methanol (ppm), C1′ is the concentration of ginsenoside Rg3 in the aqueous phase after centrifugation (ppm), and C2′ is the concentration of ginsenoside CK in the aqueous phase after centrifugation (ppm).

[0059] ② Test of centrifugal stability constant

[0060] The emulsion sample was diluted 100-fold with ultrapure water, mixed thoroughly, and the absorbance (A0) was measured at 490 nm using a UV-visible spectrophotometer. The emulsion was centrifuged at 4000 rpm for 15 minutes at 4°C. The lower layer was diluted 100-fold with ultrapure water, mixed thoroughly, and the absorbance (A0′) was measured at 490 nm using a UV-visible spectrophotometer.

[0061] The centrifugal stability constant of the emulsion sample was calculated by formula (3):

[0062]

[0063] Wherein, Ke is the centrifugal stability constant of the emulsion, A0 is the absorbance value of the emulsion dilution at 490 nm, and A0′ is the absorbance value of the lower clear liquid dilution at 490 nm.

[0064] ③ Average particle size test

[0065] The emulsion sample was diluted 300 times with ultrapure water, and the average particle size of the emulsion was measured using a particle size analyzer (the refractive indices of water and emulsion were 1.33 and 1.46, respectively).

[0066] ④Solubility test

[0067] Dissolve ginsenoside microcapsules in ultrapure water at 40°C for 10 minutes and centrifuge at 3200 rpm for 10 minutes. Take the supernatant and dry it in a 105°C oven to a constant weight (the difference between two weighings is less than 0.0005g). Record the mass of the sample after drying.

[0068] The solubility of ginsenoside microcapsules was calculated by formula (4):

[0069]

[0070] Where, S is the solubility of ginsenoside microcapsules (%), m is the mass of the sample after drying the supernatant (kg), α is the mass of ginsenoside microcapsules (kg), and ρ w The density of ultrapure water at 40°C (kg / m 3 ), V is the volume of ultrapure water when ginsenoside microcapsules are dissolved (m 3 ), m0 is the mass of the supernatant (kg), and W is the moisture content of the ginsenoside microcapsules (kg water / kg wet matter).

[0071] ⑤Wettability test

[0072] 1 g of ginsenoside microcapsules were dispersed in 100 mL of ultrapure water, and the time (min) required for all samples to pass through the water surface (or become wet) was recorded, which was the wettability of the ginsenoside microcapsules.

[0073] ⑥Dispersion test

[0074] 1 g of ginsenoside microcapsules was added to 100 mL of ultrapure water, stirred with a glass rod for 15 seconds, and then passed through a 180 μm mesh to obtain a solution mixture. 1 mL of the solution mixture was taken and dried at 108°C for 4 hours.

[0075] The dispersibility of ginsenoside microcapsules was calculated by formula (5):

[0076]

[0077] Wherein, D is the dispersibility of ginsenoside microcapsules (%), m a is the mass of ginsenoside microcapsules (kg), W is the moisture content of ginsenoside microcapsules (kg water / kg wet matter), m1 is the mass of the dried matter (kg), and m2 is the mass of the solution passing through a 180 μm sieve (kg).

[0078] ⑦Test of the release level of free fatty acids during intestinal digestion

[0079] During the intestinal stage, due to the action of lipase, the triglycerides and diglycerides contained in the oil in the sample are converted into free fatty acids. The release of free fatty acids will cause the pH value of the sample to decrease. At the time points of 20, 40, 60, 80, 100, and 120 minutes, 0.25M NaOH was added to the intestinal digestion solution to maintain the pH value of the digestion solution at 7.0. The volume of NaOH added at each time point was recorded, and the total volume of NaOH required to maintain the pH value of the intestinal digestion solution at 7.0 was calculated to calculate the lipid digestibility of the sample. The total amount of free fatty acids released was calculated using formula (6):

[0080]

[0081] Wherein, R is the total amount of free fatty acids released at each time point during the intestinal digestion stage (%), C is the concentration of NaOH (0.25 mol / L), V is the volume of NaOH added during intestinal digestion (L), M is the relative molecular mass of sunflower oil (876.2 g / mol), and m is the total mass of sunflower oil in the sample (g).

[0082] ⑧Testing the bioaccessibility of ginsenosides Rg3 and CK during intestinal digestion

[0083] 2 mL of intestinal digestion product was mixed with 8 mL of methanol and sonicated for 30 minutes. The mixture was then centrifuged at 10,000 rpm at 4°C for 30 minutes. The supernatant was filtered through a 0.45 μm organic membrane to remove impurities. The concentrations of ginsenosides Rg3 (C1) and CK (C2) in the filtrate were determined by high-performance liquid chromatography. Each sample was run in triplicate.

[0084] The intestinal digestion product was ultracentrifuged at 150,000 g for 60 minutes at 4°C. The product separated into three layers: an upper oil phase, a middle transparent micellar phase, and a bottom sediment. The middle micellar phase was filtered through a 0.45 μm organic membrane to remove impurities. The concentrations of ginsenosides Rg3 (C1′) and CK (C2′) in the filtrate were determined by high-performance liquid chromatography. Each sample was run in triplicate.

[0085] The bioaccessibility of ginsenoside Rg3 (B1) and CK (B2) refers to the ratio of the concentration of ginsenoside Rg3 and CK in micelles formed during the intestinal digestion stage to the concentration of ginsenoside Rg3 and CK in the small intestinal digestive fluid;

[0086] Calculate using formulas (7) and (8):

[0087]

[0088]

[0089] Among them, B1 is the bioaccessibility of ginsenoside Rg3 (%), B2 is the bioaccessibility of ginsenoside CK (%), C1 is the concentration of ginsenoside Rg3 in methanol (ppm), C2 is the concentration of ginsenoside CK in methanol (ppm), C1′ is the concentration of ginsenoside Rg3 in the micellar phase after ultracentrifugation (ppm), and C2′ is the concentration of ginsenoside CK in the micellar phase after ultracentrifugation (ppm).

[0090] The raw materials used in the embodiment are:

[0091] Maltodextrin: DE=17; all raw materials can be purchased commercially.

[0092] Example 1 Saponin extract containing rare ginsenosides Rg3 and CK

[0093] A method for preparing a saponin extract containing rare ginsenosides Rg3 and CK comprises the following steps:

[0094] 10 g of ginseng slices were mixed with 100 mL of citric acid solution with pH values ​​of 1.0, 2.0, or 3.0 or ultrapure water (pH value of 6.12), and then heated at 50, 70, 90, 110, or 130°C for 1, 3, or 5 h to obtain saponin extracts containing rare ginsenosides Rg3 and CK.

[0095] The obtained saponin extracts containing rare ginsenosides Rg3 and CK were tested, and the test results were as follows: Figure 1 As shown;

[0096] Statistical analysis revealed that the pH, heating temperature, and heating time of the citric acid solution interacted with the yields of ginsenosides Rg3 and CK. After heating at 50, 70, and 90°C for 1 h, the yield of ginsenoside CK increased slightly, but no significant differences were observed with decreasing pH (p>0.05). However, when the pH of the citric acid solution decreased from 2.0 to 1.0, the yield of ginsenoside Rg3 significantly increased in samples heated at 70 and 90°C. However, when heated at 110 and 130°C, the yields of ginsenosides Rg3 and CK initially increased and then decreased with decreasing pH. This may be due to the combination of extremely low pH and high temperature, which led to the decomposition or transformation of ginsenosides Rg3 and CK. After heating at 50, 70, and 90°C for 3 and 5 h, the yields of ginsenosides Rg3 and CK continued to increase with decreasing pH, with higher yields at pH 1.0 and 2.0. However, it is worth noting that when the heating temperature is above 110°C, the yield of ginsenoside Rg3 gradually decreases as the pH decreases, while the yield of ginsenoside CK first increases and then decreases. When the heating temperature is ≤90°C, pH plays a major role in increasing the yield of ginsenosides Rg3 and CK, while lower pH values ​​favor further conversion of Rg3 and CK. When the heating temperature is ≥110°C, the benefits of low pH values ​​are reduced. When heated at 130°C, the lower pH value actually leads to a decrease in the yield of ginsenosides Rg3 and CK.

[0097] from Figure 1It can be seen that the acid hydrolysis conditions for the highest yield of ginsenoside Rg3 were: pH 6.12, heating at 110°C for 3 hours; the acid hydrolysis conditions for the highest yield of ginsenoside CK were: pH 2.0, heating at 110°C for 3 hours. The optimal acid hydrolysis conditions were: pH 6.12, heating at 110°C for 3 hours; and pH 2.0, heating at 110°C for 3 and 5 hours. Ginsenosides Rg3 and CK under these conditions showed no significant differences from their respective highest yields (2.57±0.29 mg / g and 2.80±0.23 mg / g).

[0098] Example 2 Emulsion containing rare ginsenosides Rg3 and CK

[0099] A method for preparing an emulsion containing rare ginsenosides Rg3 and CK comprises the following steps:

[0100] (1) Water phase:

[0101] WPI was dissolved in water, heated at 80°C for 15 minutes, and then immediately cooled to room temperature in an ice-water bath. Tween 80 was then added, and the mixture was stirred at 200 rpm for 2 hours. Finally, the mixture was allowed to stand at 4°C for 12 hours until it was completely hydrated to obtain an aqueous phase. The ratio of WPI, Tween 80, and water was 0.5 g: 0-0.7 g: 50 mL.

[0102] (2) Oil phase:

[0103] The saponin extract containing rare ginsenosides Rg3 and CK (pH 2.0, heated at 110°C for 3 h) and sunflower oil were mixed at a volume ratio of 4:1 and stirred at 200 rpm for 12 h to obtain an oil phase;

[0104] (3) Emulsion:

[0105] The aqueous phase and the oil phase were mixed in a volume ratio of 1:1 to a total volume of 150 mL, and sheared and dispersed at a speed of 15,000 rpm for 5 minutes to obtain a crude emulsion; then the crude emulsion was microfluidized at a pressure of 600 bar for 10 minutes to obtain an emulsion containing rare ginsenosides Rg3 and CK.

[0106] The obtained emulsion was subjected to performance testing, and the test results are as follows:

[0107] The entrapment efficiency of ginsenosides Rg3 and CK and the centrifugal stability constant of the emulsion are the most important indicators for evaluating emulsion performance. The entrapment efficiency of ginsenosides Rg3 and CK is an indicator of emulsion encapsulation effectiveness, while the centrifugal stability constant is a commonly used indicator for evaluating emulsion stability during storage. For emulsion delivery systems, higher active substance entrapment efficiency and centrifugal stability constant indicate better emulsion performance. Average particle size is an indicator of emulsion stability; smaller average particle size indicates more stable emulsions.

[0108] Table 2 shows the performance test results of emulsions containing rare ginsenosides Rg3 and CK. As shown in Table 2, for emulsions prepared using WPI and Tween 80 as a combined emulsifier, the entrapment efficiency of ginsenosides Rg3 and CK and the centrifugal stability constant of the emulsions continuously increased with increasing Tween 80 dosage, then stabilized. The average particle size of the emulsions continuously decreased, then stabilized. Emulsions prepared using WPI alone exhibited a lower centrifugal stability constant and a larger average particle size. When the ratio of WPI, Tween 80, and ultrapure water was 0.5 g: 0.5-0.7 g: 50 mL, the emulsions exhibited smaller average particle sizes, higher entrapment efficiency (Rg3: 81.21-81.34%; CK: 83.31-83.44%), and stronger stability (53.13-53.56%).

[0109] Table 2 Performance test results of oil-in-water emulsion

[0110]

[0111] Note: Different letters for the same indicator indicate significant differences (p<0.05).

[0112] Example 3 Microcapsules

[0113] A method for improving the resolubility of ginsenoside microcapsules and the bioaccessibility of ginsenosides Rg3 and CK, comprising the following steps:

[0114] A mixture of 20 g of maltodextrin and inulin was used as the wall material (the mass ratio of maltodextrin to inulin was 1:3). The wall material was added to 100 mL of an emulsion containing rare ginsenosides Rg3 and CK, stirred at 200 rpm for 2 h, and after uniform mixing, ginsenoside microcapsules were obtained by spray drying. The spray drying process parameters were as follows: inlet temperature of 180 ° C, outlet temperature of 90 ° C, and air injection rate of 15 m / s. 3 / h, and the injection rate was 5mL / min.

[0115] Comparative Example 1

[0116] The inulin in Example 3 was omitted, the mass ratio of maltodextrin to inulin was 4:0, and the other conditions were the same as in Example 3 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0117] Comparative Example 2

[0118] The mass ratio of maltodextrin to inulin in Example 3 was adjusted to 3:1, and the other conditions remained the same as in Example 3 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0119] Comparative Example 3

[0120] The mass ratio of maltodextrin to inulin in Example 3 was adjusted to 2:2, and the other parameters remained the same as in Example 3 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0121] Comparative Example 4

[0122] Maltodextrin in Example 3 was omitted, and the dosage ratio of maltodextrin to inulin was 0:4. Other conditions remained the same as in Example 3 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0123] Example 4 Microcapsules

[0124] A method for improving the resolubility of ginsenoside microcapsules and the bioaccessibility of ginsenosides Rg3 and CK, comprising the following steps:

[0125] A mixture of 20 g of maltodextrin and lactose was used as the wall material (the mass ratio of maltodextrin to lactose was 1:3). The wall material was added to 100 mL of an emulsion containing rare ginsenosides Rg3 and CK, stirred at 200 rpm for 2 h, and after uniform mixing, ginsenoside microcapsules were obtained by spray drying. The spray drying process parameters were as follows: inlet temperature of 180 ° C, outlet temperature of 90 ° C, and air injection rate of 15 m / s. 3 / h, and the injection rate was 5mL / min.

[0126] Comparative Example 5

[0127] The mass ratio of maltodextrin and lactose in Example 5 was adjusted to 3:1, and the other conditions remained the same as in Example 5 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0128] Comparative Example 6

[0129] The mass ratio of maltodextrin and lactose in Example 5 was adjusted to 2:2, and the other conditions remained the same as in Example 5 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0130] Comparative Example 7

[0131] Maltodextrin in Example 5 was omitted, the mass ratio of maltodextrin to lactose was 0:4, and the other conditions were the same as in Example 5 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0132] The obtained ginsenoside microcapsules containing rare ginsenosides Rg3 and CK were subjected to a resolubility test, and the test results are as follows:

[0133] Solubility, wettability, and dispersibility are important indicators of the resolubility of microcapsules. Solubility is the ability of a powder to dissolve in a solution, wettability is the ability of a powder to absorb water and rehydrate without stirring, and dispersibility is the ability to disperse in water without agglomeration. The greater the solubility, the better the wettability (the shorter the wetting time), the stronger the dispersibility, and the better the resolubility of the microcapsules.

[0134] Table 3 shows the results of resolubility testing of ginsenoside microcapsules. Table 3 shows that for microcapsules prepared with a composite wall material of maltodextrin and inulin / lactose, solubility increases, wetting time decreases, and dispersibility improves with increasing inulin / lactose dosage. Microcapsules prepared with maltodextrin alone as the wall material exhibit lower solubility, longer wetting time, and poorer dispersibility. When the ratio of maltodextrin to inulin / lactose was 1:3, and the ratio of the wall material to the emulsion containing rare ginsenosides Rg3 and CK was 20 g:100 mL, the microcapsules exhibited higher solubility (88.58-92.10%), better wettability (0.91-1.72 min), and stronger dispersibility (89.87-91.15%), demonstrating excellent resolubility.

[0135] Table 3 Redissolution performance test results of ginsenoside microcapsules

[0136] Example Solubility (%) Wettability (min) Dispersibility (%) Example 3 88.58 ± 3.91 ab ]] <![CDATA[0.91±0.08 fg ]]> <![CDATA[91.15±0.56 a ]]> Comparative Example 1 <![CDATA[83.10±2.59 b ]]> <![CDATA[6.82±0.24 a ]]> 78.52 ± 1.39 f ]] Comparative Example 2 <![CDATA[86.30±3.74 b ]]> <![CDATA[4.06±0.09 b ]]> 84.96 ± 1.52 e ]] Comparative Example 3 <![CDATA[87.85±3.34 ab ]]> <![CDATA[2.31±0.07 d ]]> <![CDATA[88.30±1.68 bcd ]]> Comparative Example 4 <![CDATA[86.84±1.92 b ]]> <![CDATA[3.28±0.34 bc ]]> 86.45 ± 0.64 de ]] Example 4 <![CDATA[92.10±1.44 a ]]> 1.72 ± 0.08 e ]] <![CDATA[89.87±1.43 abc ]]> Comparative Example 5 <![CDATA[86.51±3.76 b ]]> <![CDATA[4.44±0.16 b ]]> <![CDATA[84.05±1.47 e ]]> Comparative Example 6 <![CDATA[89.60±1.32 ab ]]> <![CDATA[2.76±0.19 c ]]> <![CDATA[87.59±0.92 cd ]]> Comparative Example 7 87.32 ± 1.75 b ]] <![CDATA[3.69±0.46 b ]]> <![CDATA[85.72±1.73 de ]]>

[0137] Note: Different letters for the same indicator indicate significant differences (p<0.05).

[0138] The wettability test of the ginsenoside microcapsules prepared in Examples 3, 4 and Comparative Example 1 was carried out. The results of the wettability test at 2s and 30s were as follows. Figure 2 As shown;

[0139] from Figure 2 It can be seen that during the wetting process, the microcapsules continuously redissolved over time, and the turbidity of the solution continued to increase. At 2 seconds, the microcapsules prepared with maltodextrin alone as the wall material had a slower sinking rate, while the microcapsules prepared with maltodextrin and inulin / lactose (1:3) as the composite wall material had a faster sinking rate. At 30 seconds, the turbidity of the redissolved solution of the microcapsules prepared with maltodextrin alone was lower, and many visible particles were present in the solution. The turbidity of the redissolved solution of the microcapsules prepared with maltodextrin and inulin / lactose (1:3) as the composite wall material was higher, with very few visible particles. This shows that the microcapsules prepared with maltodextrin and inulin / lactose (1:3) as the composite wall material redissolved into an emulsion more quickly and had better wettability.

[0140] Example 5 Reconstituted Emulsion

[0141] 20g of ginsenoside microcapsules were added to 100mL of ultrapure water and stirred at 200rpm for 2h to obtain a ginsenoside reconstituted emulsion containing rare ginsenosides Rg3 and CK. The performance of the ginsenoside reconstituted emulsion containing rare ginsenosides Rg3 and CK was tested, and the test results are as follows:

[0142] Table 4 shows the performance test results of ginsenoside reconstituted emulsions. As shown in Table 4, for reconstituted emulsions prepared with maltodextrin and inulin / lactose as the composite wall material, the entrapment efficiency of ginsenosides Rg3 and CK and the centrifugal stability constant of the emulsions increased with increasing inulin / lactose dosage, while the average particle size of the emulsions decreased. For reconstituted emulsions prepared with maltodextrin alone as the wall material, the entrapment efficiency of ginsenosides Rg3 and CK was lower, the centrifugal stability constant was lower, and the average particle size was larger. When the dosage ratio of maltodextrin and inulin / lactose is 1:3, and the dosage ratio of wall material to emulsion containing rare ginsenosides Rg3 and CK is 20 g:100 mL, the average particle size of the reconstituted emulsion is smaller, with a higher encapsulation rate (Rg3: 70.24-73.99%; CK: 76.77-77.66%) and stronger stability (7.06-8.78%).

[0143] Table 4 Performance test results of ginsenoside reconstituted emulsion

[0144] Example Rg3 Embedding Rate (%) CK Embedding Rate (%) Centrifugal Stability Constant (%) Average Particle Size (nm) Example 3 <![CDATA[73.99±0.81 a ]]> <![CDATA[77.66±0.18 a ]]> <![CDATA[7.06±0.26 b ]]> <![CDATA[574.39±8.73 f ]]> Comparative Example 1 57.94 ± 0.35 f ]] <![CDATA[67.54±0.53 g ]]> <![CDATA[1.16±0.05 f ]]> <![CDATA[1131.94±36.03 a ]]> Comparative Example 2 <![CDATA[61.88±1.34 e ]]> 73.35 ± 0.18 e ]] <![CDATA[2.33±0.25 e ]]> <![CDATA[891.72±8.13 b ]]> Comparative Example 3 <![CDATA[68.90±0.40 d ]]> <![CDATA[75.64±0.12 b ]]> <![CDATA[3.94±0.13 c ]]> <![CDATA[724.41±6.69 d ]]> Comparative Example 4 <![CDATA[67.83±1.72 d ]]> 74.92 ± 0.33 bc ]] <![CDATA[3.61±0.26 d ]]> <![CDATA[753.91±15.89 c ]]> Example 4 <![CDATA[70.24±0.92 bc ]]> 76.77 ± 0.87 a ]] 8.78 ± 0.10 a ]] 494.37 ± 7.20 g ]] Comparative Example 5 <![CDATA[62.00±1.25 e ]]> <![CDATA[71.95±0.22 f ]]> <![CDATA[1.38±0.11 f ]]> <![CDATA[745.43±12.36 cd ]]> Comparative Example 6 <![CDATA[68.08±0.87 d ]]> <![CDATA[74.44±0.53 cd ]]> <![CDATA[4.03±0.04 c ]]> <![CDATA[632.82±8.96 e ]]> Comparative Example 7 <![CDATA[67.04±1.09 d ]]> <![CDATA[74.13±0.42 d ]]> <![CDATA[3.97±0.49 cd ]]> <![CDATA[716.08±10.50 d ]]>

[0145] Note: Different letters for the same indicator indicate significant differences (p<0.05).

[0146] Example 6 In vitro digestion of reconstituted emulsion

[0147] The ginsenoside microcapsules prepared in Examples 3 and 4 and Comparative Example 1 were stirred at 200 rpm for 4 h to form a reconstituted emulsion;

[0148] The reconstituted emulsion was subjected to simulated oral digestion, simulated gastric digestion and simulated intestinal digestion in an in vitro digestion experimental model (see the literature C.Liu, T.Yang, ZKZhao, T.Liu, KXLi, JGLiu, P.Zhou, Effects ofparticle size reduction combined with beta-cyclodextrin on the in vitrodissolution and in vivo relative bioavailability of ginsenosides in Panaxginseng[J].Food&Function, 2022, 13(21): 10882-10894) to obtain products of each stage of in vitro simulated digestion.

[0149] The products of each stage of in vitro simulated digestion were subjected to performance testing, and the test results are as follows:

[0150] Table 5 shows the particle size results of the products of the reconstituted ginsenoside emulsion at each stage of in vitro simulated digestion. As can be seen from Table 5, microcapsules prepared with maltodextrin alone as the wall material produced larger particle sizes and poorer stability at each stage of the in vitro simulated digestion process. However, microcapsules prepared with a composite wall material of maltodextrin:inulin / lactose (1:3) produced smaller particle sizes and better stability at each stage of the in vitro simulated digestion process.

[0151] Table 5 Particle size results of products at each stage of in vitro simulated digestion of ginsenoside reconstituted emulsion

[0152]

[0153] Note: Different letters for the same indicator indicate significant differences (p<0.05).

[0154] The results of the release level of free fatty acids during the intestinal digestion stage are as follows Figure 3 As shown. Figure 3It can be seen that during the intestinal digestion phase, the free fatty acid release of each reconstituted emulsion group increased with prolonged digestion time, eventually reaching a plateau. The reconstituted emulsion formed by microcapsules prepared with a maltodextrin:inulin ratio of 1:3 exhibited the highest free fatty acid release rate between 0 and 80 minutes, followed by microcapsules prepared with a maltodextrin:lactose ratio of 1:3, and the lowest free fatty acid release rate from microcapsules prepared with maltodextrin alone. The final free fatty acid release from reconstituted emulsions prepared with a maltodextrin:inulin ratio of 1:3 was the highest, followed by microcapsules prepared with a maltodextrin:lactose ratio of 1:3, and the lowest free fatty acid release rate from microcapsules prepared with maltodextrin alone.

[0155] The bioaccessibility results of ginsenosides Rg3 and CK are as follows Figure 4 As shown. Figure 4 As can be seen, the bioaccessibility of ginsenosides Rg3 and CK was highest in the reconstituted emulsion formed by microcapsules prepared with a maltodextrin:inulin ratio of 1:3 as the wall material, followed by that formed by microcapsules prepared with a maltodextrin:lactose ratio of 1:3, and lowest in the reconstituted emulsion formed by microcapsules prepared with maltodextrin alone as the wall material. During intestinal digestion, bile salts can partially or completely displace Tween 80 surrounding oil droplets, promoting the binding of lipase / lipase complexes at the oil-water interface, leading to the progressive hydrolysis of triglycerides, ultimately producing monoglycerides and free fatty acids. Medium- and short-chain monoglycerides and free fatty acids are released into the surrounding intestinal fluid, while long-chain monoglycerides and free fatty acids, due to their higher surface activity, may accumulate at the oil-water interface. The presence of digestion products at the interface can prevent pancreatic lipase from accessing triglycerides and inhibit lipid digestion. Bile salts and phospholipids can incorporate monoglycerides and free fatty acids at the oil-water interface into micelles and vesicles, producing mixed micelles for delivering nutrients such as ginsenosides. The function of mixed micelles is to deliver ginsenosides Rg3 and CK. The higher the amount of free fatty acids released, the more mixed micelles are formed, and the greater the amount of ginsenosides Rg3 and CK that can be delivered. In other words, the final amount of free fatty acids released is positively correlated with the bioaccessibility of ginsenosides.

[0156] Comparative Example 8

[0157] The inulin in Example 3 was adjusted to β-cyclodextrin, the mass ratio of maltodextrin to β-cyclodextrin was 1:3, and the other conditions were kept consistent with Example 3 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0158] Comparative Example 9

[0159] The maltodextrin in Example 3 was adjusted to β-cyclodextrin, and the mass ratio of β-cyclodextrin to inulin was 1:3. Other conditions remained the same as in Example 3 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0160] Comparative Example 10

[0161] The maltodextrin in Example 5 was adjusted to β-cyclodextrin, and the mass ratio of β-cyclodextrin to lactose was 1:3. Other conditions remained the same as in Example 5 to prepare ginsenoside microcapsules containing rare ginsenosides Rg3 and CK.

[0162] The obtained ginsenoside microcapsules containing rare ginsenosides Rg3 and CK were subjected to a resolubility test, and the test results are as follows:

[0163] Table 6 shows the resolubility test results of ginsenoside microcapsules prepared with different wall materials. As can be seen from Table 6, the solubility, wettability, and dispersibility of microcapsules prepared by adjusting maltodextrin (or inulin, lactose) to β-cyclodextrin were poor, inferior to those prepared with maltodextrin and inulin / lactose.

[0164] Table 6 Redissolution performance test results of ginsenoside microcapsules

[0165] Example Solubility (%) Wettability (min) Dispersibility (%) Example 3 <![CDATA[88.58±3.91 ab ]]> <![CDATA[0.91±0.08 e ]]> <![CDATA[91.15±0.56 a ]]> Example 4 <![CDATA[92.10±1.44 a ]]> <![CDATA[1.72±0.08 d ]]> 89.87 ± 1.43 a ]]> Comparative Example 8 <![CDATA[81.27±2.10 c ]]> <![CDATA[6.93±0.53 a ]]> <![CDATA[77.86±2.15 d ]]> Comparative Example 9 <![CDATA[84.65±1.32 bc ]]> <![CDATA[3.08±0.32 c ]]> <![CDATA[86.83±1.39 b ]]> Comparative Example 10 85.39 ± 2.49 bc ]]> <![CDATA[4.15±0.47 b ]]> <![CDATA[84.23±0.86 c ]]>

[0166] Note: Different letters for the same indicator indicate significant differences (p<0.05).

[0167] In summary, the present invention screened many components and found that: only in the technical scheme of the present invention, when maltodextrin and inulin / lactose are used as composite wall materials, the dosage ratio of maltodextrin and inulin / lactose is 1:2-4, and the dosage ratio of the wall material to the emulsion containing rare ginsenosides Rg3 and CK is 15-30g:100mL, the ginsenoside microcapsules have good solubility, wettability and dispersibility, and their re-dissolved emulsion has a high ginsenoside Rg3 and CK embedding rate and strong stability. If other types of wall materials are selected, such as maltodextrin and β-cyclodextrin, β-cyclodextrin and inulin, and β-cyclodextrin and lactose, the re-dissolution performance (solubility, wettability and dispersibility) of the ginsenoside microcapsules are poor. When the addition amount is not in accordance with the present invention, such as when the ratio of maltodextrin to inulin / lactose is 3:1 and 2:2, the re-dissolution performance of the ginsenoside microcapsules is worse than that of the optimal component; and the results of Comparative Example 1 show that without the addition of inulin / lactose, the re-dissolution performance of the ginsenoside microcapsules is poor; the results of Comparative Examples 4 and 7 show that without the addition of maltodextrin, the re-dissolution performance of the ginsenoside microcapsules is poor.

[0168] Therefore, from the above examples and comparative examples, it can be seen that the ginsenoside microcapsules prepared by using malt dextrin and inulin / lactose as the composite wall material have good solubility, wettability and dispersibility, the reconstituted emulsion has high embedding rate of ginsenoside Rg3 and CK and strong stability. The amount ratio of malt dextrin and inulin / lactose is 1:2-4, and the amount ratio of the wall material to the emulsion containing rare ginsenoside Rg3 and CK is 15-30 g:100 mL, which is a key technical means and has great market promotion value.

[0169] Although the present application has been disclosed with reference to the preferred embodiments, it is not intended to limit the present application, and any person skilled in the art can make various modifications and modifications without departing from the spirit and scope of the present application, and therefore the protection scope of the present application should be defined by the claims.

Claims

1. A method for improving the resolubility of ginsenoside microcapsules and the bioaccessibility of ginsenosides Rg3 and CK, characterized in that: The steps include: The wall material and the emulsion containing rare ginsenosides Rg3 and CK were mixed uniformly and then spray-dried to obtain ginsenoside microcapsules; The dosage ratio of the wall material and the emulsion containing rare ginsenosides Rg3 and CK was 20 g:100 mL; The wall material is a mixture of maltodextrin and lactose in a mass ratio of 1:3, or a mixture of maltodextrin and inulin in a mass ratio of 1:3; The preparation method of the emulsion containing rare ginsenosides Rg3 and CK comprises the following steps: (1) Water phase: Mix WPI, Tween 80 and water in the ratio of 0.5 g: 0.5-0.7 g: 50 mL to obtain an aqueous phase; (2) Oil phase: The saponin extract containing rare ginsenosides Rg3 and CK and edible oil are mixed uniformly to obtain an oil phase; (3) Emulsion: The aqueous phase and the oil phase are mixed and shear-dispersed to obtain a coarse emulsion; the coarse emulsion is then homogenized by microfluidization to obtain an emulsion containing rare ginsenosides Rg3 and CK.

2. The method according to claim 1, characterized in that The process parameters of the spray drying are: inlet temperature of 150-190℃, outlet temperature of 80-100℃, air injection rate of 10-30m 3 / h, and the injection rate was 2-10mL / min.

3. The method according to claim 1, characterized in that The volume ratio of the water phase to the oil phase is 1:

1.

4. Ginsenoside microcapsules prepared by the method according to any one of claims 1 to 3.

5. A ginsenoside microcapsule reconstituted emulsion, characterized in that: The emulsion is obtained by adding the ginsenoside microcapsules according to claim 4 into water and mixing them evenly.

6. Use of the ginsenoside microcapsule reconstituted emulsion according to claim 5 in the field of functional foods.

7. Use of the ginsenoside microcapsules according to claim 4 in the field of functional foods.

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