Bird's nest acid liposome composition and preparation method thereof

Abstract

The invention provides a sialic acid liposome composition and a preparation method thereof. The preparation method of the sialic acid liposome composition comprises the following steps: adding an organic phase containing phospholipid and ethanol into a water phase containing sialic acid and water, and uniformly mixing; the water phase also comprises maltodextrin. The sialic acid lipidosome composition with high bioavailability, high stability and good application performance is prepared by the preparation method provided by the invention, and the sialic acid lipidosome composition is simple in preparation process, capable of realizing large-scale industrialization, low in cost and relatively high in commercial application value.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of food, daily chemical, and more particularly, to a bird's nest acid liposome composition and a preparation method thereof. BACKGROUND

[0002] Bird's nest acid is increasingly used in the fields of daily chemical and food due to its excellent efficacy. The existing technology applies bird's nest acid in a free / unwrapped manner in the field of food, which has problems of gastrointestinal degradation and low bioavailability. For example, CN115997840A discloses a whitening and anti-aging functional ice cream based on the addition of bird's nest acid and a preparation method thereof, CN115624132A discloses a bird's nest acid eight-treasure congee and a preparation method thereof, CN113974144A discloses a instant bird's nest acid meal replacement powder, and CN111248373A discloses a bird's nest acid beverage and a preparation method thereof. In the above patent application files, bird's nest acid is consumed in an unwrapped / free form, which has problems of gastrointestinal degradation, difficulty of polar molecules to penetrate the lipid bilayer membrane, first-pass effect, and the like, resulting in low bioavailability of bird's nest acid.

[0003] Liposome drug delivery technology has been widely used in the fields of medicine, food, daily chemical, and the like due to its excellent biocompatibility and safety, sustained and controlled release, bioavailability, and the like. There are few studies on the wrapping of bird's nest acid using liposome technology, and there are problems of simple system, poor flexibility of liposome structure, poor stability, and low bioavailability. For example, CN114099410A discloses a face cream composition with synergistic gain anti-aging efficacy liposome and a preparation method thereof, which uses a thin film dispersion method to prepare bird's nest acid liposome. The process flow is complex, the cost is high, commercial application is difficult, and the components of the liposome include phospholipid, bacillus licheniformis / glutamic acid fermentation filtrate, bird's nest acid, proanthocyanidin solution, and phosphate buffer solution without other additives. The liposome structure has problems of poor flexibility, poor stability, and low bioavailability. In summary, it is urgent to develop a bird's nest acid liposome system with high bioavailability and high stability. SUMMARY

[0004] In order to solve or at least partially solve the problems in the prior art, the present application provides a bird's nest acid liposome composition and a preparation method thereof. The bird's nest acid liposome composition provided by the present application has high bioavailability and good stability.

[0005] The first object of the present application is to provide a preparation method of a bird's nest acid liposome composition, which comprises the following steps: An organic phase containing phospholipid and ethanol is added to an aqueous phase containing bird's nest acid and water, and mixed uniformly. The aqueous phase further includes maltodextrin.

[0006] The application prepares bird's nest acid into a liposome dosage form and modifies the surface of the liposome using maltodextrin as a stabilizer, protects the structure stability of the liposome during the drying process, improves the bioavailability of bird's nest acid, and significantly improves the stability of the bird's nest acid liposome, effectively solving the problems of low bioavailability of bird's nest acid and poor stability of conventional bird's nest acid liposome in the prior art.

[0007] In a preferred embodiment of the application, in order to improve the reconstitution stability of the bird's nest acid liposome, especially the liposome composition, and improve the encapsulation efficiency, the bird's nest acid is 40-60 parts by weight, the phospholipid is 3-10 parts by weight, the maltodextrin is 20-40 parts by weight, and the ethanol is 3-20 parts by weight. In the application, the amount of water is conventionally selected, and in a specific embodiment, the amount of water can be 150-300 parts.

[0008] In the application, as the amount of phospholipid increases, the particle size of the obtained bird's nest acid liposome increases, and the absolute value of the potential increases. When the amount of phospholipid is 3-10 parts, the particle size, Pdi, and potential of the dried liposome are basically unchanged, the reconstitution performance of the bird's nest acid liposome is good, and the encapsulation efficiency gradually increases. The amount of phospholipid can be 3-10 parts, 6-10 parts, or 8-10 parts. In a preferred embodiment of the application, in order to further improve the encapsulation efficiency, the amount of phospholipid is 8-10 parts. In the application, the phospholipid can be selected from commonly used phospholipids in the art, and the source of the phospholipid is not limited in the scheme given in the application. In a specific embodiment, sunflower phospholipid is only used as an example.

[0009] In a preferred embodiment of the application, in order to further improve the stability of the bird's nest acid liposome, reduce the particle size of the bird's nest acid liposome, and improve the encapsulation efficiency and bioavailability, glycerol can be added to the aqueous phase in the above preparation method. At the same time, it is found that the larger the amount of glycerol, the smaller the particle size of the bird's nest acid liposome, and the better the stability of the obtained liposome. However, when the amount is too large, the obtained bird's nest acid liposome will be sticky, so the amount of glycerol is 0-12 parts by weight (based on 40-60 parts of bird's nest acid), which can be 2-12 parts by weight, and is preferably 5-12 parts by weight.

[0010] In the detailed embodiments of the present application, the organic phase and the aqueous phase can be mixed uniformly by using methods commonly used in the art, such as shearing, stirring and / or high-pressure homogenization. In the detailed embodiments of the present application, the organic phase and the aqueous phase are usually prepared separately, and then the organic phase is added to the aqueous phase under shearing, and the system is sheared until uniform to form a liposome primary solution. The liposome primary solution is subjected to high-pressure homogenization to obtain a liposome (homogenized) solution, sterilization and drying. In this step, the temperature of the aqueous phase is 50-60°C. The shearing conditions after the organic phase is added to the aqueous phase are as follows: the shearing temperature is 50-60°C, the shearing time is 10-20 min, and the shearing speed is 5000-8000 rpm / min. In the preparation of the organic phase, phospholipid and ethanol can be mixed uniformly to obtain the organic phase by using methods commonly used in the art, such as shearing the phospholipid and ethanol until the system is uniform in an optional embodiment. In this process, the shearing temperature is 30-50°C, the shearing time is 5-8 min, and the shearing speed is 5000-8000 rpm / min. In the preparation of the aqueous phase, bird's nest acid and other raw materials can be mixed uniformly to obtain the aqueous phase by using methods commonly used in the art, such as shearing the raw materials of the aqueous phase until the system is uniform in an optional embodiment. In this process, the shearing temperature is 30-50°C, the shearing time is 10-20 min, and the shearing speed is 5000-8000 rpm / min.

[0011] In the detailed embodiments of the present application, the homogenization pressure is 60-80 MPa, and the homogenization frequency is 2-3 times.

[0012] In the present application, the bird's nest acid liposome composition can be a solution, a powder or the like, and the bird's nest acid liposome composition of the present application can be prepared into a desired dosage form according to the needs of the person skilled in the art. When the bird's nest acid liposome composition is required to be a powder, the uniformly mixed composition can be dried. In the detailed embodiments of the present application, the drying can be freeze-drying / spray-drying. The steps of freeze-drying can be pre-freezing at -80°C for 8-24 h, and then freeze-drying for 24-48 h. The steps of spray-drying can be an inlet air temperature of 160-170°C and a material temperature of 65-70°C. After drying, the step of sterile packaging is usually included. The person skilled in the art can select whether to sterilize according to the needs.

[0013] Another object of the present application is to provide a bird's nest acid liposome composition prepared by the above preparation method.

[0014] Still another object of the present application is to provide a bird's nest acid liposome composition comprising, by weight, 40-60 parts of bird's nest acid, 3-10 parts of phospholipid, 20-40 parts of maltodextrin and 0-12 parts of glycerol. The phospholipid is preferably 8-10 parts. The glycerol is preferably 5-12 parts.

[0015] Another object of the present invention is to provide a sanilic acid liposome powder comprising the above-mentioned sanilic acid liposome composition. In the preparation method of the liposome powder, both ethanol and water are removed, and the resulting sanilic acid liposome powder comprises 40-60 parts of sanilic acid, 3-10 parts of phospholipids, and 20-40 parts of maltodextrin. If, in a preferred embodiment, glycerol is added to the raw materials, the liposome powder also contains glycerol. Other preferred technical solutions are not further described.

[0016] The swalate acid liposomes provided by the present invention have a nano-liposome structure (redissolved particle size 100-350nm, potential -10--30mV), no solvent residue, good stability (stability index of homogeneous liquid <1), high bioavailability and good application performance.

[0017] The present invention provides a new slant acid liposome composition, which produces a slant acid liposome with high bioavailability and high stability, while also having good application performance. By preparing slant acid into a liposome dosage form, its bioavailability is significantly improved compared to the free form (reducing gastrointestinal degradation and improving the bioaccessibility of the active ingredient; the liposomes can enhance absorption efficiency through transmembrane transport and lymphatic absorption; the liposomes' sustained and controlled release mechanisms prolong the in vivo circulation time). By using liposome surface modification technology, the stability of the slant acid liposomes is significantly improved, expanding their scope of application and filling market and technological gaps in this field. The present invention produces a slant acid liposome composition that combines high bioavailability with good stability and good application performance. It has the advantages of simple process, large-scale industrialization, low cost, and high commercial application value. DETAILED DESCRIPTION

[0018] Below in conjunction with embodiment, specific embodiment of the present invention is described in further detail.The following examples are used to illustrate the present invention, but are not used to limit the scope of the invention.In the present invention, unless otherwise specified, "%" is mass percentage, and the raw material dosage and content related in the present invention are all weight ratios.In the present invention, mass parts can be weight units well known in the art such as μg, mg, g, kg, etc., and can also be multiples thereof, such as 1 / 10, 1 / 100, 10 times, 100 times, etc.

[0019] The preparation method of the swan nest acid liposome powder provided in the embodiments and comparative examples of the present invention comprises the following steps: S1: Weigh sunflower lecithin and ethanol according to the formula in the table and fully shear until the system is uniform. The shear temperature is 45°C, the shear time is 6 minutes, and the shear speed is 7000 rpm / min. This is the organic phase; S2: The bird's nest acid, stabilizer, co-emulsifier and deionized water are weighed according to the formula, and are fully sheared to be uniform in the system, wherein the shearing temperature is 45 DEG C, the shearing time is 15 min, and the shearing speed is 8000 rpm / min, namely the water phase; S3: The organic phase is slowly added to the water phase at 55 DEG C ± 5 DEG C under continuous shearing, and is fully sheared to be uniform in the system, wherein the shearing temperature is 55 DEG C, the shearing time is 15 min, and the shearing speed is 8000 rpm / min, to form a liposome initial solution; S4: The liposome initial solution obtained in step S3 is subjected to high-pressure homogenization at 60 Mpa twice to obtain a liposome (homogenized) solution; S5: The liposome solution is subjected to sterilization treatment to obtain a sterilized solution; S6: The sterilized solution is subjected to spray drying to obtain a bird's nest acid liposome powder, and the spray drying conditions are as follows: the inlet air temperature is 165 ± 5 DEG C, and the material temperature is 65-70 DEG C.

[0020] Test method of the application: 1. Stability index test A stability analyzer Turbiscan Tower (France FORMULACTION Company) is used, and the stability index is obtained under the condition of 25 DEG C and for 6h; the larger the stability index, the worse the sample stability.

[0021] 2. Particle size, Pdi and potential test The liposome homogenized solution / powder is diluted with water to a suitable concentration, and a Malven Zetasizer Pro is used to test the particle size, Pdi and potential thereof.

[0022] 3. Redissolvability 2g of the powder is poured into a beaker containing water (25 DEG C, 98g of water), and the powder is observed for complete dissolution after 30s of stirring at a speed of 3 rpm / s.

[0023] 4. Encapsulation efficiency and content The bird's nest acid liposome powder is diluted 10 times with water, 0.4mL of which is taken and placed in an ultrafiltration centrifuge tube (molecular weight cut-off 10kDa), and the filtrate is collected after centrifugation at 5000 rpm / min for 15 min, and the content of the bird's nest acid in the filtrate is determined by HPLC method, and the content of free bird's nest acid WF is calculated. Another 0.4mL of the 10 times water diluent is taken, acetonitrile is added to break the emulsion, and the content of the bird's nest acid in the supernatant is determined after centrifugation, and the total content of the bird's nest acid WT is calculated. The encapsulation efficiency EE (%) = (WT-WF) / WT x 100.

[0024] 5. Stability of aqueous solution The Edible Bird's Nest Acid Liposome Powder was diluted 50 times with water and stored at 25℃ for 1 month. The changes in appearance, particle size, PDi, potential, and content of the aqueous solution before and after storage were tested. If the appearance did not change and the change in the other indicators was less than 5%, the aqueous solution was determined to be stable.

[0025] 6、Stability The Edible Bird's Nest Acid Liposome Powder was stored at 37℃, refrigerated (4℃), and room temperature (25℃) for 3 months, respectively. The changes in reconstitution, particle size, Pdi, potential, and encapsulation efficiency were evaluated. If the reconstitution did not change and the change in the other indicators was less than 5%, the Edible Bird's Nest Acid Liposome Powder was determined to be stable.

[0026] 7、Bioavailability test In vitro simulation of gastrointestinal digestion: A two-step in vitro digestion model was used to simulate the digestion of Edible Bird's Nest Acid / Edible Bird's Nest Acid Liposome Powder in the stomach and small intestine. All experiments were performed at 37℃ and all solutions were preheated to that temperature before use.

[0027] Among them, the simulated gastric juice (pH=2.0) contains 5.505 mg / mL NaCl, 1.648 mg / mL KCl, 0.532 mg / mL NaH2PO4, 0.604 mg / mL CaCl2, 0.612 mg / mL NH4Cl, 1250 U / mL pepsin, and pH is adjusted to 2.0 with HCl. 2.5 g of sample was dissolved in ultrapure water and mixed with 200 mL of simulated gastric juice, placed in a 250 mL conical flask, and shaken at 100 rpm / min in a constant temperature horizontal shaker at 37℃ for 2 h. The pH of the reaction solution was maintained at 2.0 during the reaction.

[0028] During small intestine digestion, the gastric juice was adjusted to pH 7.0 with 1 M NaHCO3. Then an equal volume of simulated intestinal fluid containing 14.024 mg / mL NaCl, 1.128 mg / mL KCl, 6.776 mg / mL NaHCO3, 0.160 mg / mL KH2PO4, 0.100 mg / mL MgCl2, 72 U / mL trypsin, and 180 U / mL lipase was added. The simulated digestion was carried out at 37℃ in a constant temperature horizontal shaker at 100 rpm / min for 2 h, cooled in ice for 10 min to stop the intestinal digestion process, and the sample was centrifuged at 10000×g at 4℃ for 10 min. The supernatant (ID) was collected and freeze-dried for subsequent content analysis.

[0029] Bioavailability (%) = Content after gastrointestinal digestion / Content before digestion × 100; The higher the bioavailability, the less degradation of Edible Bird's Nest Acid in the gastrointestinal tract and the higher the bioavailability.

[0030] 1、Stability of the stabilizer The samples (in parts by weight) were prepared according to the formula of each raw material given in Table 1, and the effects of different stabilizers on the liposome homogeneous liquid (stability index, particle size, Pdi, potential), liposome powder (powder state, reconstitution, particle size, Pdi, potential) were evaluated. Among them, composition 1 is example 1, and compositions 2-7 are comparative examples 1-6.

[0031] Table 1 ; The results of Table 1 show that when the stabilizer is ionic, the stability index of the liposome homogeneous liquid is larger (>1), the liquid stability is poor, it does not meet the requirements of freeze drying / spray drying, and the test is discontinued; when the stabilizer is a sugar syrup with a small molecular weight, the reconstitution after drying is poor, and the particle size and Pdi increase significantly, indicating that the sugar syrup has a small protective effect on the structure of the liposome during the drying process of the liposome; when maltodextrin is used as the stabilizer, the reconstitution of the liposome homogeneous liquid after drying is good, and the particle size / Pdi / potential is basically unchanged, indicating that the maltodextrin has an excellent protective effect on the structure of the liposome during the drying process of the liposome, and therefore maltodextrin is preferred, which is one of the core points of the present application. In addition, the encapsulation efficiency of composition 1 is measured to be 34.84%.

[0032] 2, Effect of Glycerol Amount on Liposomes The samples (in parts by weight) were prepared according to the formula of Table 2, and the effects of different amounts of co-emulsifier glycerol on the liposome homogeneous liquid (stability index, particle size, Pdi, potential), liposome powder (powder state, reconstitution, particle size, Pdi, potential) were evaluated. Among them, compositions 8-14 are examples 2-8.

[0033] Table 2 ; The results of Table 2 show that as the amount of co-emulsifier (glycerol) increases, the stability index of the liquid decreases, the stability of the liquid increases, the particle size of the liquid shows a decreasing trend, and the particle size of the liposome powder decreases significantly, indicating that the co-emulsifier (glycerol) significantly improves the stability of the liposome; but when the content of co-emulsifier (glycerol) is ≥15 parts, the softening point of the powder decreases, and the powder appears to be sticky; therefore, the content of co-emulsifier (glycerol) is preferably 0-12 parts. That is, when no glycerol is added, the stability index of the liquid is also good, and the particle size of the liquid can meet the requirements. In order to improve the effect, the content of glycerol is preferably 5-12 parts.

[0034] 3, Effect of Phospholipid Amount on Liposomes Sample was prepared according to the formulation of Table 3 (in parts by weight), and the effect of different phospholipid amount on the homogenate of liposome (stability index, particle size, Pdi, potential), powder (powder state, reconstitution, particle size, Pdi, potential) was evaluated. Among them, compositions 15-21 are examples 9-15.

[0035] Table 3 ; Table 3 results show that as the amount of phospholipid increases, the stability index of the homogenate increases, the stability of the homogenate decreases, the particle size of the homogenate increases, and the absolute value of the potential increases; when the amount of phospholipid is 3-10 parts, the particle size, Pdi, and potential of the dried liposome are basically unchanged, the reconstitution performance of the liposome powder is good, and the encapsulation efficiency gradually increases, which meets the expectation; when the amount of phospholipid is 13-20 parts, the particle size and Pdi of the dried liposome significantly increase, and the reconstitution performance of the liposome powder decreases, which is analyzed to be caused by the destruction of the liposome structure during the drying process; therefore, the preferred amount of phospholipid is 8-10 parts.

[0036] 4, Effect of core components on liposome Sample was prepared according to the formulation of Table 4 (in parts by weight), and the effect of each component on liposome was evaluated. Among them, composition 18 is example 11, composition 22 is example 12, and composition 23 is comparative example 7. The composition and amount of raw materials in control groups 1-2 are shown in Table 4.

[0037] Table 4 ; Table 4 results show that maltodextrin has a significant structure protection effect on the drying process of liposome, which can avoid significant increase in particle size during the drying process; among them, as a preferred, glycerol can also improve the bioavailability and encapsulation efficiency of the liposome powder. The phospholipid / edible bird's nest acid system is simple, and has the problems of poor reconstitution, large particle size, and large Pdi. For bioavailability: edible bird's nest acid / phospholipid / maltodextrin / glycerol > edible bird's nest acid / phospholipid / maltodextrin > edible bird's nest acid, the bioavailability of edible bird's nest acid liposome powder is significantly improved compared with that of edible bird's nest acid, which indicates that the liposome structure can effectively protect the structure of edible bird's nest acid in the gastrointestinal tract, reduce its degradation, and improve the bioavailability; the edible bird's nest acid liposome powder has obvious liposome characteristics, high encapsulation efficiency, and high bioavailability, and has good powder state, stability, reconstitution, and aqueous solution stability, which indicates that it has good application performance and commercial application value.

[0038] Finally, the method of the present application is only a preferred embodiment, and is not used to limit the protection scope of the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing a senna acid liposome composition, characterized in that: The steps include: The organic phase containing phospholipids and ethanol is added to the aqueous phase containing swalnic acid and water, and mixed evenly; the aqueous phase also includes maltodextrin.

2. The preparation method according to claim 1, characterized in that Calculated by weight, the ingredients include 40-60 parts of bird's nest acid, 3-10 parts of phospholipids, 20-40 parts of maltodextrin, and 3-20 parts of ethanol.

3. The preparation method according to claim 2, characterized in that The phospholipids are 8-10 parts.

4. The preparation method according to any one of claims 1 to 3, characterized in that The aqueous phase also includes glycerol.

5. The preparation method according to claim 4, characterized in that In parts by weight, the amount of glycerol used is 0 to 12 parts, preferably 5 to 12 parts.

6. The swalnic acid liposome composition obtained by the preparation method according to any one of claims 1 to 5.

7. A senna acid liposome composition, characterized in that: Calculated by weight, the invention comprises 40-60 parts of swiftlet acid, 3-10 parts of phospholipids, 20-40 parts of maltodextrin and 0-12 parts of glycerol.

8. The senna acid liposome composition according to claim 7, characterized in that The phospholipids are 8 to 10 parts.

9. The senna acid liposome composition according to claim 7, characterized in that The glycerol is 5 to 12 parts.

10. A swan acid liposome powder, characterized in that: The invention comprises the swine acid liposome composition according to any one of claims 6 to 9.