A method for improving bioavailability of phytosterols

Abstract

The present application relates to the development and utilization of phytosterol technical field, more specifically to a kind of preparation method of phytosterol effervescent tablet.The present application first prepares phytosterol nanolipid carrier suspension, then it is mixed with excipient after spray drying and is prepared the solid dispersion of phytosterol nanocarrier.Further the solid dispersion of phytosterol nanocarrier, disintegrating agent and auxiliary material are prepared phytosterol effervescent tablet;The nanometer structure lipid carrier prepared has nanometer size, good polydispersity, and the encapsulation rate can reach 96.9%.The phytosterol effervescent tablet prepared has good flavor and uniformity, convenient to take, better taste, has higher stability while can be quickly disintegrated;Bioavailability can reach 43.3%, and the content of phytosterol in effervescent tablet can reach 25.76mg / g.Overcome the problem of poor solubility and bad taste of phytosterol greatly improve the bioavailability of phytosterol;It has higher application value.

Description

Technical Field

[0001] This invention relates to the field of phytosterol development and utilization technology, and more specifically to a method for improving the bioavailability of phytosterols. Background Technology

[0002] Phytosterols, structurally similar to cholesterol, are widely recognized for their cholesterol-lowering activity. The recommended daily intake is 2 grams to reduce plasma cholesterol levels and cardiovascular disease risk. Since it is difficult to achieve sufficient daily intake of phytosterols (150-400 mg / day) from a diet, incorporating them into functional foods may be an effective way to realize their health benefits. However, the poor solubility and wettability of phytosterols in water limit their formulation in water-based products such as beverages. Secondly, the high melting point, waxy consistency, and high crystalline structure of phytosterols result in a gritty texture, leading to a final product with undesirable sensory properties. Furthermore, their low solubility and high crystallinity in water and oil lead to insufficient intestinal absorption of crude phytosterols (0.5%-2%), thus affecting their cholesterol-lowering activity.

[0003] However, the existing plant sterol products have the following disadvantages: (1) Esterification of plant sterol with lipophilic groups increases its fat solubility, and then it is dissolved in oil to make soft capsules, which will greatly increase the daily intake of oil and is not beneficial to people's health; (2) After microcrystallizing plant sterol, it is taken with water. Due to the high melting point and high crystal structure of plant sterol, it will produce a gritty texture, resulting in poor sensory characteristics and poor intestinal absorption.

[0004] Nanostructured lipid carriers (NLCs) are second-generation solid lipid nanoparticles (NPs) composed of biocompatible, biodegradable, non-sensitizing, and non-irritating lipids, prepared by mixing solid and liquid (oil) lipids. A key characteristic of NLCs is the disordered crystalline structure resulting from the high and low melting points of lipids. Due to the different properties of solid and liquid lipids, the mixed lipid matrix forms numerous imperfect crystals during cooling. These crystals possess limited order, creating additional space for encapsulating active substances.

[0005] To address the above problems, this invention proposes an oral effervescent tablet that facilitates the absorption of phytosterols and its preparation method. Summary of the Invention

[0006] This invention provides an oral effervescent tablet that facilitates the absorption of phytosterols and its preparation method, aiming to solve the problems of current phytosterol products containing a large amount of oil, having poor sensory characteristics, and leading to poor intestinal absorption.

[0007] To solve the above problems, the present invention adopts the following technical solution: First, a phytosterol nanocarrier solid dispersion is prepared by spray drying after mixing a phytosterol nanolipid carrier suspension with an excipient; in the phytosterol nanolipid carrier suspension, the ratio of phytosterol to distearate, medium-chain glycerides, and lecithin is 10:20~40:20~40:1~5; the phytosterol is selected from one of methyl sterol and dimethyl sterol.

[0008] The first aspect of the present invention provides a method for improving the bioavailability of phytosterols, comprising preparing phytosterols into effervescent tablets, the phytosterol effervescent tablets being made of the following components:

[0009] 35-50 parts of solid dispersion of phytosterol nanocarriers;

[0010] 30-40 parts disintegrant;

[0011] 0-12 parts of sodium carboxymethyl starch;

[0012] Flavoring agent 0-4 parts;

[0013] 3-6 parts lubricant;

[0014] 2-6 parts adhesive;

[0015] 0-4 parts of auxiliary agent;

[0016] Furthermore, the solid dispersion of the phytosterol nanocarrier is prepared by spray drying after mixing a phytosterol nanolipid carrier suspension with an excipient.

[0017] Furthermore, in the phytosterol nanolipid carrier, the ratio of phytosterol to distearate, medium-chain glycerides, and lecithin is 10:20~40:20~40:1~5.

[0018] In some embodiments of the present invention, the phytosterol is selected from non-methyl sterols and dimethyl sterols;

[0019] In some embodiments of the present invention, the additives include magnesium stearate, food flavoring, and micronized silica gel.

[0020] Furthermore, the disintegrant includes an acid source and an alkaline source;

[0021] In some embodiments of the present invention, the flavoring agent is selected from aspartame, sucrose and lecithin;

[0022] In some embodiments of the present invention, the lubricant is selected from PEG4000, PEG6000, L-leucine and sodium dodecyl sulfate;

[0023] In some embodiments of the present invention, the adhesive is selected from PVP (polyvinylpyrrolidone), ethanol, and water, etc.

[0024] A second aspect of the present invention is to provide a method for preparing the phytosterol effervescent tablets, comprising the following steps:

[0025] S11. Preparation of acid particles: The solid dispersion of phytosterol nanocarrier, sodium carboxymethyl starch and acid source material are mixed evenly, and then an appropriate amount of binder is added. After wetting, the mixture is granulated to obtain acid particles.

[0026] S12. Preparation of alkali granules: Mix the alkali source material, magnesium stearate, and food flavoring evenly, then add an appropriate amount of binder, moisten, and granulate to obtain alkali granules;

[0027] S13. Tableting and coating: After drying the acid granules and alkali granules separately at 40-60℃, they are mixed and lubricant, micronized silica gel and magnesium stearate are added. After mixing, the mixture is compressed into tablets. A water-soluble starch coating is applied to the surface and dried to obtain the effervescent tablet product.

[0028] Furthermore, the preparation method of the phytosterol nanocarrier solid dispersion includes the following steps:

[0029] S21. Preparation of the organic phase: Distearate, medium-chain triglycerides, and lecithin are dissolved in a mixed solvent of ethanol / acetone and heated until fully dissolved to form the organic phase;

[0030] S22. Preparation of aqueous phase: Take Tween 80 solution and heat it to the same temperature as the organic phase to obtain the aqueous phase;

[0031] S23. Preparation of suspension: The organic phase is slowly injected into the aqueous phase under stirring using an injection device, then homogenized under high pressure, and placed under heating to allow the solvent to evaporate. Then chitosan solution is added and stirred, followed by sodium tripolyphosphate solution and stirring. The resulting translucent emulsion is placed under an ice bath and stirred to obtain a phytosterol nanostructure lipid carrier suspension.

[0032] S24. Preparation of suspension: Add the excipient maltodextrin to the suspension obtained in step S23, and obtain a solid dispersion of nanostructured lipid carrier by spray drying. After sieving, the solid dispersion of nanocarrier is obtained.

[0033] In some embodiments of the present invention, the acid source is selected from one or a combination of tartaric acid, fumaric acid or citric acid.

[0034] In some embodiments of the present invention, the alkaline source is selected from one or a combination of sodium bicarbonate and sodium urethane.

[0035] Further, in step S21, an ethanol / acetone mixed solvent is used, wherein the ratio of ethanol / acetone is 1:5 to 5:1, preferably 1:5 to 2:3;

[0036] Furthermore, the ratio of the phytosterol to the mixed solvent is 1 g: 30~50 ml;

[0037] In some embodiments of the present invention, in step S11, the temperature is raised to 60-75°C;

[0038] In some embodiments of the present invention, in step S22, the concentration of the Tween 80 solution is 0.5-3%, preferably 1-3%;

[0039] Furthermore, the mass ratio of the phytosterol to the volume of the organic phase is 1 g: 50~200 ml.

[0040] In some embodiments of the present invention, in step S23, the solvent evaporation step is heated to 60~65°C.

[0041] In some embodiments of the present invention, in step S23, the ratio of the amount of chitosan solution to the volume of the aqueous phase is 1:1, and the concentration of chitosan in the solution is 0.1~2 wt%.

[0042] In some embodiments of the present invention, in step S23, the ratio of sodium tripolyphosphate solution to water phase is 1:100 to 1:2, and the concentration of sodium tripolyphosphate is 1 to 10 wt%.

[0043] In some embodiments of the present invention, in step S24, the amount of maltodextrin used is 0.5 to 2 times the total weight of phytosterols, distearate, medium-chain triglycerides and lecithin.

[0044] In some embodiments of the present invention, in step S24, the spray drying conditions are: inlet air temperature of 170°C, outlet air temperature of 80-85°C, air volume of 120 m³ / min, and feed rate of 600 ml / h.

[0045] The beneficial effects of the technical solution of this invention are at least as follows:

[0046] (1) In this invention, phytosterols are first encapsulated in a nanostructured lipid carrier. The resulting nanostructured lipid carrier has a nanoscale size, good aggregation and dispersibility, and an encapsulation rate of up to 96.9%.

[0047] (2) The present invention further prepares the nanostructured lipid carrier into a solid dispersion by spray drying, which effectively expands its application scenarios;

[0048] (3) The present invention prepares phytosterol effervescent tablets based on solid dispersions; the preparation process adopts acid-base separation granulation method, which effectively prevents the acid source and the base source from reacting prematurely during the preparation process, so that the effervescent tablets can disintegrate quickly when dissolved and release the active ingredients more quickly; the effervescent tablets have high stability and can disintegrate quickly, which greatly improves the bioavailability of phytosterols; its phytosterol content can reach 25.76 mg / g;

[0049] (4) The phytosterol effervescent tablets prepared by this invention have good flavor and uniformity, are convenient to take and have a good taste; they greatly improve the bioavailability of phytosterols by overcoming the problems of poor solubility and unpleasant taste; the bioavailability can reach 43.3%. Attached Figure Description

[0050] The accompanying drawings are provided to further illustrate the present application and form part of the specification. They are used together with the embodiments of the present application to explain the present application, but do not constitute a limitation thereof.

[0051] Figure 1 This is a flowchart of the preparation process of the present invention. Detailed Implementation

[0052] To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the examples in the specification. Many specific details are set forth in the following description to provide a thorough understanding of the present invention; however, the present invention may also be implemented in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0053] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0054] Quality evaluation standards for phytosterol nanostructured lipid carriers

[0055] (1) Particle size potential measurement

[0056] A suitable amount of phytosterol nanostructured lipid carrier was taken, and its particle size and potential were measured using a Malvernb Zeta potentiometer.

[0057] (2) Encapsulation efficiency determination

[0058] Take an appropriate amount of phytosterol nanostructured lipid carrier, add 10 mL of n-hexane, and immediately vortex for 5 min. Then allow the solution to stand for 1-2 min to dissolve the unencapsulated bioactive substance. Collect the supernatant hexane and dry it with nitrogen. Redissolve the residue in anhydrous ethanol, add ferric phosphate reagent to the redissolved residue solution, react for 15 min, and then place 200 µl in an ELISA plate. Measure the absorbance of the bioactive substance at a specific absorption wavelength using an ELISA reader. Calculate the bioactive substance content based on the standard curve (R² > 0.99) established for the purified bioactive substance. The encapsulation efficiency formula is as follows:

[0059]

[0060] In the formula: W 总生物活性物质 It is the mass (mg) of the added bioactive substance, W 游离生物活性物质 It is the mass (mg) of bioactive substances that are not encapsulated in the nanostructured lipid carrier and are free in solution. It is the mass (mg) of the lipid phase formed by solid lipids and liquid lipids.

[0061] Quality evaluation standards for phytosterol solid dispersion effervescent tablets

[0062] (1) Weight difference

[0063] Take 20 tablets of the test sample, accurately weigh the total weight, calculate the average tablet weight, and then accurately weigh each tablet separately. Compare the weight of each tablet with the average tablet weight, and calculate the limit of tablet weight difference according to the following formula.

[0064]

[0065] The results must comply with the Pharmacopoeia of the People's Republic of China (General Chapter 0101, 2020 edition).

[0066] (2) Disintegration time limit

[0067] Take one tablet and place it in a 250 ml beaker (containing 200 ml of water at 20℃ ± 5℃). Numerous bubbles will be released. When the gas escapes from the tablet or fragments stops, the tablet should dissolve or disperse in the water without any aggregated particles remaining. Test six tablets using the same method; each tablet should disintegrate within 5 minutes. Results must comply with the *Pharmacopoeia of the People's Republic of China* (General Chapter 0901, 2020 edition).

[0068] (3) Friability test

[0069] Take 10 tablets, blow away the powder that has fallen off the tablets with a hair dryer, weigh them accurately, and place them in a tablet friability tester and rotate them 100 times. Take them out, remove the powder in the same way, weigh them accurately, and the weight loss of the effervescent tablets should be less than 1%, with no breakage, cracking or pulverization, in accordance with the "Pharmacopoeia of the People's Republic of China" (General Chapter 0923, 2020 edition).

[0070] Example 1

[0071] Preparation of phytosterol nanostructured lipid carriers and their solid dispersion effervescent tablets

[0072] Step 1: Preparation of phytosterol nanostructured lipid carriers

[0073] Take 10 parts of soybean sterol, 30 parts of distearate (solid lipid), 30 parts of medium-chain triglycerides (liquid lipid), 2 parts of lecithin, 100 parts of anhydrous ethanol, and 400 parts of acetone in a beaker and heat in a 65°C constant temperature water bath until fully dissolved to form the organic phase; separately take 1000 parts of 2% Tween 80 solution and heat in a water bath to the same temperature as the organic phase to form the aqueous phase. Slowly inject the organic phase into the aqueous phase with a needle while stirring at 1000 r / min, stir at (65±2)°C, then homogenize under high pressure at 500 bar, and place it in a 65°C water bath for 2 h to allow the organic solvent to evaporate. Then add 1000 parts of 5 mg / ml chitosan solution and stir for 1 h, then add 200 parts of 50 mg / ml sodium tripolyphosphate solution and stir for 3 h. Place the resulting translucent emulsion in an ice bath and stir to obtain a phytosterol nanostructure lipid carrier suspension.

[0074] Step 2: Preparation of solid dispersions of phytosterol nanocarriers

[0075] Add 100 parts of maltodextrin as an excipient to the nanostructured lipid carrier suspension prepared above, and obtain a solid dispersion of the nanostructured lipid carrier by spray drying. Pass the dispersion through a 100-mesh sieve to obtain the solid dispersion of the nanocarrier. The spray drying conditions are: inlet air temperature 170℃, outlet air temperature 80-85℃, and air volume of... The feed rate is 600 ml / h.

[0076] Step 3: Preparation of solid dispersion effervescent tablets

[0077] The formulation of an oral effervescent tablet that facilitates the absorption of phytosterols comprises: 40 parts of a solid dispersion of phytosterol nanocarriers (including 20 parts of maltodextrin), 35 parts of disintegrant, 11 parts of sodium carboxymethyl starch, 2.5 parts of flavoring agent (aspartame), 5 parts of lubricant (PEG6000:NaCl=2:3), 4 parts of binder (10% PVP anhydrous ethanol solution), 1 part of micronized silica gel, 0.5 parts of magnesium stearate, and 1 part of edible flavoring. The disintegrant is derived from tartaric acid and sodium bicarbonate. All excipients are sieved through a 100-mesh sieve. The specific preparation process is as follows:

[0078] S1. Preparation of acid particles: Mix the solid dispersion of the nanocarrier encapsulating phytosterols in the prescribed amount with tartaric acid, then add an appropriate amount of binder (10% PVP anhydrous ethanol solution), moisten it and then granulate to obtain acid particles.

[0079] S2. Preparation of alkaline granules: Mix sodium bicarbonate, aspartame and food flavoring in the prescribed amounts evenly, then add an appropriate amount of binder (10% PVP anhydrous ethanol solution), moisten and granulate to obtain alkaline granules.

[0080] S3, tableting: Dry the acid granules and alkali granules separately at 40-60℃ for 10 min, mix them, add lubricant (PEG6000:NaCl=2:3), micronized silica gel, and magnesium stearate, mix well, and compress into effervescent tablets.

[0081] S4. Coating: Coat the surface of the effervescent tablet semi-finished product with a layer of water-soluble starch coating, dry it, and obtain the effervescent tablet finished product.

[0082] Example 2

[0083] The difference from Example 1 is that the formulation of the effervescent tablets in the third step is changed to: 47 parts of solid dispersion of phytosterol nanocarrier (including 23.5 parts of maltodextrin), 39 parts of disintegrant, 2.5 parts of flavoring agent (aspartame), 5 parts of lubricant (PEG6000:NaCl=2:3), 4 parts of binder (10% PVP anhydrous ethanol solution), 1 part of micronized silica gel, 0.5 parts of magnesium stearate, and 1 part of edible flavoring. The disintegrant is provided by tartaric acid as the acid source and sodium bicarbonate as the alkali source.

[0084] Example 3

[0085] The difference from Example 1 is that the formulation of the effervescent tablets in the third step is changed to: 40 parts of solid dispersion of phytosterol nanocarrier (including 20 parts of maltodextrin), 12.5 parts of sodium carboxymethyl starch, 35 parts of disintegrant, 2.5 parts of flavoring agent (aspartame), 5 parts of lubricant (PEG6000:NaCl=2:3), 4 parts of binder (10% PVP anhydrous ethanol solution) and 1 part of food flavoring, wherein the acid source of the disintegrant is tartaric acid and the alkali source is sodium bicarbonate.

[0086] Example 4

[0087] The difference from Example 1 is that the acid source in the effervescent tablet formulation in the third step is changed to citric acid.

[0088] Example 5

[0089] The difference from Example 1 is that the ratio of phytosterols to distearate, medium-chain glycerides and lecithin is changed to 10:20:40:2.

[0090] Example 6

[0091] The difference from Example 1 is that the ratio of phytosterols to distearate, medium-chain triglycerides and lecithin is changed to 10:40:20:2.

[0092] Example 7

[0093] The difference from Example 1 is that the heating temperature during the solvent evaporation stage is 60°C.

[0094] Example 8

[0095] The difference from Example 1 is that the concentration of the Tween 80 solution is changed to 3%.

[0096] Comparative Example 1: Phytosterol Nanostructured Lipid Carrier Emulsion

[0097] The phytosterol nanostructure lipid carrier emulsion was prepared according to the first step of Example 1.

[0098] Comparative Example 2: Phytosterol Emulsion

[0099] Take 2.4 parts of soybean oil, heat it to 60℃, and then add 0.4 parts of phytosterol and stir evenly to form a lipid phase; add 2 parts of Tween 80 to 100 parts of water, heat and mix at 60℃ to obtain an aqueous phase; then mix the aqueous phase and lipid phase, and shear at 12000 rpm for 3 min to obtain an oil-in-water pre-emulsion; homogenize the oil-in-water pre-emulsion three times under high pressure at 500 bar, and cool to obtain a phytosterol emulsion.

[0100] Comparative Example 3

[0101] The difference from Comparative Example 1 is that the ratio of phytosterols to distearate, medium-chain triglycerides and lecithin was changed to 15:30:30:2.

[0102] Comparative Example 4

[0103] The difference from Example 1 is that the ratio of ethanol to acetone in the organic phase is 2:3 and 1:1, respectively.

[0104] Comparative Example 5

[0105] The difference from Example 1 is that the concentration of the Tween 80 solution is changed to 1%.

[0106] Comparative Example 6

[0107] The difference from Example 1 is that the heating temperature during the solvent evaporation stage is 70°C and 75°C.

[0108] Comparative Example 7

[0109] The difference from Example 1 is that step S4 in the third step is not present; the surface of the effervescent tablet is not coated with water-soluble starch.

[0110] The properties of phytosterol nanostructured lipid carriers are shown in the table below;

[0111] Table 1 Properties of nanostructured lipid carriers

[0112]

[0113] As shown in Table 1, with the increase of phytosterol addition, the particle size increases and the encapsulation efficiency decreases. Simultaneously, with the increase of the solid lipid / liquid lipid ratio, the particle size also increases, but the PDI decreases. This may be because with the increase of liquid lipid content, the formed nanocarriers become more irregular, leading to a less homogeneous and less stable system.

[0114] Example 1 and Comparative Example 4 investigated the effect of the ethanol / acetone ratio in the organic phase on the properties of the nanocarrier. As the ethanol content increased, the heating time for evaporating the organic solvent became longer. The longer heating time led to the destruction of the nanocarrier, resulting in an increase in both the particle size and PDI of the nanocarrier.

[0115] The data from Examples 1, 8, and 5 were compared to investigate the effect of the amount of surfactant added in the aqueous phase on the properties of the nanocarrier. The nanocarrier system formed by a 1% concentration of Tween 80 solution was unstable, with both the particle size and PDI being relatively large; while the system formed by a 3% concentration of Tween 80 solution was stable, but the particle size of the nanocarrier was slightly increased.

[0116] Example 1 and Comparative Example 6 investigated the effect of heating temperature on the properties of nanocarriers during solvent evaporation. As the heating temperature increased, the particle size of the nanocarriers increased. This may be because the nanocarrier structure disintegrated and was destroyed as the heating temperature increased, resulting in a certain degree of aggregation.

[0117] Experimental Example 1: Determination of Phytosterol Content in Phytosterol Solid Dispersion Effervescent Tablets

[0118] Take 10 effervescent tablets, accurately weigh them, grind them finely, and accurately weigh a certain weight (approximately equivalent to 0.1g of phytosterol). Dissolve them in 100 ml of purified water. Take 300 µl of the dissolved phytosterol and add 5 ml of methanol. Sonicate in a water bath for 1 min to break the emulsion. Then add 5 ml of n-hexane and vortex to mix. Extract the phytosterol from the methanol phase twice. Combine the n-hexane phases and dry them under nitrogen. Finally, redissolve the sample to 2 mL with the mobile phase. Add ferric phosphate reagent to the redissolved residue solution and react for 15 min. Take 200 µl of the solution and place it in an ELISA plate. Measure the absorbance of the phytosterol using an ELISA reader at a specific phytosterol absorption wavelength. Calculate the phytosterol content based on the standard curve (R2>0.99) established for purified phytosterol.

[0119] Table 2 Quality Inspection

[0120]

[0121] Quality inspection of the samples revealed that the effervescent tablets had a longer disintegration time without the addition of sodium carboxymethyl starch. This may be because the granules produced when using only dextrin as a filler were harder, affecting the disintegration time. However, when sodium carboxymethyl starch was added as a co-filler, it could quickly absorb water and achieve significant swelling, thereby accelerating the disintegration rate. Furthermore, the disintegration time was also relatively longer when the acid source was changed from tartaric acid to citric acid. This may be because tartaric acid is a stronger acid than citric acid, which allows it to better provide an acidic environment and accelerate the disintegration rate. In addition, without the addition of magnesium stearate and micronized silica gel, tablet sticking occurred during compression, resulting in significant differences in the weight of the effervescent tablets.

[0122] Sensory analysis of Experiment Example 2

[0123] The sensory analysis included sample form (for effervescent tablets, the form after brewing), homogeneity, and flavor. Each item was scored out of 10. The sensory analysis was conducted by 20 professionals. After each set of samples was tested, the sample rinsed with purified water, and the next set of samples was tested at least 2 hours apart. The sensory score was the total score of all items. The higher the score, the closer the sample was to the product's optimal characteristics. The sensory score results are shown in Table 2.

[0124] Table 3 Sensory Analysis Results

[0125]

[0126] Experimental Example 3: In vitro digestion assay for bioaccessibility

[0127] 1. Test Methods

[0128] (1) Phytosterol solid dispersion effervescent tablets

[0129] Oral digestion stage: Accurately weigh 10 tablets of this product, grind them into a fine powder, accurately weigh 16.0 g, add 100 ml of purified water to dissolve (to achieve a lipid content of 2% (w / w)), mix with an equal volume of preheated oral working solution (80 ml + 80 ml), adjust to pH = 6.80, and shake at 100 rpm for 2 min at 37°C to obtain the oral digestion solution. The oral working solution contains 30 mg / mL mucin and needs to be dissolved by stirring at 4°C for 12 hours in advance.

[0130] Gastric digestion stage: The simulated gastric juice, which has been preheated at 37°C, is mixed with an equal volume of oral digestive juice (100 ml + 100 ml), the pH is adjusted to 2.50, and the mixture is incubated at 37°C and 100 rpm for 2 h to obtain the gastric digestive juice.

[0131] Small intestinal digestion stage: 60 mL of gastric digestive fluid was taken and the pH adjusted to 6.99. Then, 3 mL of preheated (37°C) small intestinal stock solution and 7 mL of bile salt solution (53.6 mg / mL) were added sequentially. After readjusting the pH to 6.99, 5 mL of pancreatic lipase solution (24 mg / mL, lipase activity 64~256 U / mL) was rapidly added to initiate intestinal digestion. The reaction was carried out at 37°C for 2 h to simulate the small intestinal digestion stage. During the reaction, 0.1 mol / L NaOH was continuously added dropwise using an automatic titrator to neutralize free fatty acids (FFA) and maintain the pH at 6.995~6.999.

[0132] The micelle phase was obtained by centrifuging the enzyme-inactivated small intestinal digestive fluid at 12000 g for 60 min at 4°C. A small amount of undigested fat was visible in the upper layer, followed by a clear intermediate phase and a white precipitate in the lower phase. The clear intermediate liquid was aspirated with a syringe; this was the micelle phase. Phytosterols in the micelle phase were extracted with n-hexane, and the PS content was measured. PS bioaccessibility was calculated using the following formula:

[0133]

[0134] in The concentration of PS in the micelle phase. This represents the concentration of PS in the small intestinal products.

[0135] 2. Test Results

[0136] Table 3 shows the results of the bioavailability of phytosterols in the phytosterol solid dispersion effervescent tablets. It can be seen that the bioavailability of phytosterols in the phytosterol solid dispersion effervescent tablets is significantly greater than that in the control group. This indicates that encapsulating phytosterols in a nanostructured lipid carrier and drying it into a solid dispersion to make effervescent tablets greatly improves the bioavailability of phytosterols while overcoming their poor solubility and unpleasant taste, achieving the expected results.

[0137] Table 4 Results of Biological Accessibility Test

[0138]

[0139] This invention first prepares a phytosterol nanolipid carrier suspension, then mixes it with excipients and spray-dries it to obtain a solid dispersion of phytosterol nanocarriers. Further, the solid dispersion of phytosterol nanocarriers, disintegrants, and excipients are used to prepare phytosterol effervescent tablets. The obtained nanostructured lipid carriers have nanoscale dimensions, good aggregation and dispersibility, and an encapsulation efficiency of up to 96.9%. The obtained phytosterol effervescent tablets have good flavor and homogeneity, good sample transparency, and no obvious gritty feeling or special flavor; they are convenient to take, have a good taste, and exhibit high stability while rapidly disintegrating; the bioavailability can reach 43.3%, and the phytosterol content in the effervescent tablets can reach 25.76 mg / g. This invention overcomes the problems of poor solubility and unpleasant taste of phytosterols, greatly improving their bioavailability; it has high application value.

[0140] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the present invention.

Claims

1. A method for improving the bioavailability of phytosterols, characterized in that, Phytosterols are formulated into effervescent tablets, which are made from the following components: 35-50 parts of solid dispersion of phytosterol nanocarriers; 30-40 parts disintegrant; 0-12 parts of sodium carboxymethyl starch, wherein the amount of sodium carboxymethyl starch used is not 0; Flavoring agent 0-4 parts; 3-6 parts lubricant; 2-6 parts adhesive; 0-4 parts of the auxiliary agent, and the amount of the auxiliary agent is not 0; The solid dispersion of the phytosterol nanocarrier is prepared by spray drying after mixing a phytosterol nanolipid carrier suspension with an excipient. In the phytosterol nanolipid carrier, the ratio of phytosterol to distearate, medium-chain glycerides and lecithin is 10:20~40:20~40:1~5. The additives are magnesium stearate, food flavoring, and micronized silica gel. The disintegrant is an acid source and an alkaline source, and the acid source is tartaric acid. The preparation method of the phytosterol effervescent tablets includes the following steps: S11. Preparation of acid particles: The solid dispersion of phytosterol nanocarrier, sodium carboxymethyl starch and acid source material are mixed evenly, and then an appropriate amount of binder is added. After wetting, the mixture is granulated to obtain acid particles. S12. Preparation of alkali granules: Mix the alkali source material, magnesium stearate, and food flavoring evenly, then add an appropriate amount of binder, moisten, and granulate to obtain alkali granules; S13. Tableting and coating: After drying the acid granules and alkali granules separately at 40-60℃, they are mixed and lubricant, micronized silica gel and magnesium stearate are added. After mixing, the mixture is compressed into tablets. A water-soluble starch coating is applied to the surface and dried to obtain the effervescent tablet product.

2. The method for improving the bioavailability of phytosterols according to claim 1, characterized in that, The phytosterols are selected from non-methyl sterols and dimethyl sterols; And / or, the flavoring agent is selected from aspartame, sucrose and lecithin; And / or, the lubricant is selected from PEG4000, PEG6000, L-leucine and sodium dodecyl sulfate; And / or, the adhesive is selected from PVP, ethanol and water.

3. The method for preparing the phytosterol effervescent tablets according to any one of claims 1-2, characterized in that, Includes the following steps: S11. Preparation of acid particles: The solid dispersion of phytosterol nanocarrier, sodium carboxymethyl starch and acid source material are mixed evenly, and then an appropriate amount of binder is added. After wetting, the mixture is granulated to obtain acid particles. S12. Preparation of alkali granules: Mix the alkali source material, magnesium stearate, and food flavoring evenly, then add an appropriate amount of binder, moisten, and granulate to obtain alkali granules; S13. Tableting and coating: After drying the acid granules and alkali granules separately at 40-60℃, they are mixed and lubricant, micronized silica gel and magnesium stearate are added. After mixing, the mixture is compressed into tablets. A water-soluble starch coating is applied to the surface and dried to obtain the effervescent tablet product.

4. The method for preparing phytosterol effervescent tablets according to claim 3, characterized in that, The preparation method of the phytosterol nanocarrier solid dispersion includes the following steps: S21. Preparation of the organic phase: Distearate, medium-chain triglycerides, and lecithin are dissolved in a mixed solvent of ethanol / acetone and heated until fully dissolved to form the organic phase; S22. Preparation of aqueous phase: Take Tween 80 solution and heat it to the same temperature as the organic phase to obtain the aqueous phase; S23. Preparation of suspension: The organic phase is slowly injected into the aqueous phase under stirring using an injection device, then homogenized under high pressure, and placed under heating to allow the solvent to evaporate. Then chitosan solution is added and stirred, followed by sodium tripolyphosphate solution and stirring. The resulting translucent emulsion is placed under an ice bath and stirred to obtain a phytosterol nanostructure lipid carrier suspension. S24. Preparation of suspension: Add the excipient maltodextrin to the suspension obtained in step S23, and obtain a solid dispersion of nanostructured lipid carrier by spray drying. After sieving, the solid dispersion of nanocarrier is obtained.

5. The method for preparing phytosterol effervescent tablets according to claim 3, characterized in that, The alkaline source is selected from sodium bicarbonate.

6. The method for preparing phytosterol effervescent tablets according to claim 4, characterized in that, In step S21, the ratio of ethanol to acetone in the ethanol / acetone mixed solvent is 1:5-2:3; the ratio of the amount of phytosterol to the amount of the mixed solvent is 1g:30~50 ml.

7. The method for preparing phytosterol effervescent tablets according to claim 4, characterized in that, In step S22, the concentration of Tween 80 solution is 0.5-3%, and the mass ratio of the phytosterol to the volume of the organic phase is 1 g: 50-200 ml.

8. The method for preparing phytosterol effervescent tablets according to claim 4, characterized in that, In step S23, the solvent evaporation step involves heating to 60~65℃.

9. The method for preparing phytosterol effervescent tablets according to claim 4, characterized in that, In step S23, the ratio of the amount of chitosan solution to the volume of the aqueous phase is 1:1, and the concentration of chitosan in the solution is 0.1~2 wt%.

10. The method for preparing phytosterol effervescent tablets according to claim 4, characterized in that, In step S23, the ratio of sodium tripolyphosphate solution to water phase is 1:100 to 1:2, and the concentration of sodium tripolyphosphate is 1 to 10 wt%.

11. The method for preparing phytosterol effervescent tablets according to claim 4, characterized in that, In step S24, the amount of maltodextrin used is 0.5 to 2 times the total weight of phytosterols, distearate, medium-chain triglycerides and lecithin.

12. The method for preparing phytosterol effervescent tablets according to claim 4, characterized in that, In step S24, the spray drying conditions are: inlet air temperature 170℃, outlet air temperature 80-85℃, and air volume 120 m³ / h. 3 / min, feed rate is 600 ml / h.

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