Manufacturing method of liposomalization

Abstract

The present invention relates to a method for manufacturing liposomal products, and more specifically, comprises: a first step of mixing a nonionic surfactant and soybean lecithin to produce a phospholipid dispersion and an oil phase forming product; a second step of adding a liposomal product to the phospholipid dispersion and oil phase forming product and mixing to form a reaction mixture; and a third step of setting a stirring speed, temperature, and time and carrying out a liposomal reaction in the reaction mixture; wherein the mixing ratio (w / w) of the soybean lecithin to the nonionic surfactant is 20:1 to 5, and the first step of producing the phospholipid dispersion and oil phase forming product may be solvent-free.

Description

Technology Field

[0001] The present invention relates to a method for manufacturing liposomal structures that facilitates industrial-scale production. Background Technology

[0002] A liposome refers to a lipid vesicle (i.e., an artificial membrane). Molecules such as phospholipids possess both a hydrophobic (water-repelling) and a hydrophilic (water-attracting) portion within a single molecule. When such a molecule is placed in water, the hydrophilic portion attempts to approach the water, while the hydrophobic portion avoids it and aggregates with other hydrophobic parts. As a result, a lipid bilayer structure is formed, which is called a liposome. Liposomes composed of phospholipids can be found within cells, and surfactants can also be used to form liposomes. Liposomes are mostly spherical or oval in shape, and because they contain internal spaces, they can be used to encapsulate functional substances for in vivo delivery. The advantages of liposomes include excellent biocompatibility and biodegradability. Although they are widely used due to these benefits, they have the disadvantage of reduced storage stability because they are sensitive to external environmental factors (e.g., temperature, pH, light, oxidation, etc.), leading to aggregation, precipitation, and layer separation over time. Liposomes vary in shape and size depending on the manufacturing method and are classified accordingly.

[0003] The size ranges widely from approximately 20 nm to 1,000 nm, and they can be manufactured in various sizes ranging from nanometers to micrometers. Liposome manufacturing technologies based on structure can be classified into single-membrane liposome manufacturing, multilayer liposome manufacturing, and multi-liquid crystal liposome manufacturing. Stable, industrial-scale liposome production is difficult because the production methods and equipment used vary depending on the target material. Furthermore, there is the challenge of having to conduct individual research and development for the liposomalization of specific target materials. The problem to be solved

[0004] According to one embodiment, the present invention provides a method for manufacturing liposomal substances capable of industrial-scale production by improving the process for liposomalization of a target substance to solve the aforementioned problems, thereby increasing process efficiency and standardizing the entire process.

[0005] However, the problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below. means of solving the problem

[0006] A method for producing liposomalization according to one embodiment comprises: a first step of mixing a nonionic surfactant and soybean lecithin to produce a phospholipid dispersion and an oil phase forming product; a second step of adding a liposomalization target substance to the phospholipid dispersion and an oil phase forming product and mixing to form a reaction mixture; and a third step of setting a stirring speed, temperature, and time and carrying out a liposomalization reaction in the reaction mixture; wherein the mixing ratio (w / w) of the soybean lecithin to the nonionic surfactant is 20:1 to 5, and the first step of producing the phospholipid dispersion and an oil phase forming product may be solvent-free.

[0007] According to one embodiment, the nonionic surfactant is introduced as a nonionic surfactant complex composed of a single nonionic surfactant or two or more nonionic surfactants; the nonionic surfactant is a liquid nonionic surfactant comprising polyoxyethylene sorbitan ester, and the mixing ratio (w / w) of the first nonionic surfactant to the remaining surfactants in the nonionic surfactant complex may be 3:1 to 2.

[0008] According to one embodiment, the first nonionic surfactant among the nonionic surfactant complex is Tween 20, and the remaining nonionic surfactants may comprise one or more selected from the group consisting of Tween 40, Tween 60, Tween 65, Tween 80, and Tween 85.

[0009] According to one embodiment, the liposomal target substance is a functional food ingredient, and the functional food ingredient may be an animal or plant extract; an animal or plant concentrate; or a vitamin.

[0010] According to one embodiment, the functional food ingredient may be red ginseng concentrate, maca extract, or vitamin C.

[0011] According to one embodiment, the mixing ratio (based on weight) of the liposomal target substance to the soybean lecithin may be 1:1 to 3.

[0012] According to one embodiment, the liposomal target material is introduced together with purified water, and the mixing ratio (w / w) of the liposomal target material to the purified water may be 1:1.5 to 6.

[0013] According to one embodiment, the reaction mixture may comprise 24% to 40% by weight of soybean lecithin, 1% to 3% by weight of a nonionic surfactant, 10% to 25% by weight of a liposomal target substance, and 35% to 64% by weight of purified water, and may further comprise 1% to 3% by weight of a reducing sugar.

[0014] According to one embodiment, the first step may be stirring for 5 to 10 minutes at a stirring speed of 50 rpm to about 200 rpm.

[0015] According to one embodiment, the third step may comprise a three-stage stirring process in which the stirring time and stirring speed are continuously increased, and the three-stage stirring process may include: a first stirring step of stirring at 100 rpm to 500 rpm for 0 to 5 minutes; a second stirring step of stirring at 600 rpm to 1,500 rpm for 5 to 15 minutes; and a third stirring step of stirring at 1,600 rpm to 2,000 rpm for 15 to 30 minutes.

[0016] According to one embodiment, the second stirring step may be 5°C to 20°C higher than the temperature of the first stirring step, and the third stirring step may be 5°C to 10°C higher than the temperature of the second stirring step.

[0017] According to one embodiment, the liposomal manufacturing method may provide liposomal particles with a size of 300 nm (nanometer) to 2,000 nm (nanometer). Effects of the invention

[0018] The present invention can improve process efficiency by improving the liposomalization process (e.g., process steps, process time, etc.) and classify materials into several categories, thereby providing a standardized liposomalization manufacturing method suitable for each material.

[0019] In addition, the liposomal manufacturing method according to the present invention has the advantage of enabling the liposomalization of health functional food ingredients on an industrial scale and improving product performance by increasing the absorption rate of the health functional food ingredients. Brief explanation of the drawing

[0020] FIG. 1 is a process flow diagram of the liposomal manufacturing method of the present invention according to embodiments. Specific details for implementing the invention

[0021] Hereinafter, the liposomal manufacturing method of the present invention will be described in detail with reference to the examples and drawings. However, the present invention is not limited to these examples and drawings.

[0022] FIG. 1 is an exemplary process flow diagram of a method for manufacturing liposomalization according to one embodiment of the present invention. In FIG. 1, the method for manufacturing liposomalization may include: a first step (100) of mixing a nonionic surfactant and soybean lecithin to produce a phospholipid dispersion and an oil phase forming product; a second step (200) of adding and mixing a liposomalization target substance to the phospholipid dispersion and the oil phase forming product; and a third step (300) of setting a stirring speed, temperature, and time and proceeding with a liposomalization reaction.

[0023] According to one embodiment, the first step (100) of preparing a phospholipid dispersion and an oil phase formation product may be performed by mixing a nonionic surfactant and soybean lecithin in a solvent-free environment (e.g., organic solvent) to form the phospholipid dispersion and oil phase formation product. That is, in order to disperse the phospholipid by a physical method without using an organic solvent in the phospholipid dispersion and oil phase formation step, an oil phase formation can be prepared according to the rotational speed and time of a rotating shaft.

[0024] According to one embodiment, a phospholipid dispersion and an oil phase forming product can be prepared by mixing a nonionic surfactant and soybean lecithin in a mixing container at a stirring speed of about 50 rpm to about 200 rpm; about 50 rpm to about 150 rpm; or about 50 rpm to about 100 rpm for 5 minutes to 10 minutes; or about 5 minutes to 8 minutes. According to one embodiment, the first step (100) can be performed at room temperature (e.g., RT, or 20 ℃ to 25 ℃). That is, the first step (100) is prepared by a physical method of mixing a nonionic surfactant and soybean lecithin without adding a solvent to produce a phospholipid dispersion and an oil phase formation, thereby making it easy to set and control appropriate process conditions (e.g., mixing ratio, stirring speed, reaction temperature, etc.) for the liposomal reaction according to the characteristics of the target substance to be liposomed, and allowing for a stable and efficient liposomal reaction to proceed by responding quickly and appropriately to various target substances.

[0025] According to one embodiment, in the first step (100), the mixing ratio (based on weight) of the soybean lecithin to the nonionic surfactant may be 20:1 to 3; 20:1 to 2; or 20:1 to 1.5. By applying the above mixing ratio, a stable and efficient liposomal reaction can be carried out.

[0026] According to one embodiment, the soybean lecithin is isolated and extracted by filtering and hydrogenating soybean oil, and is used in food processing, pharmaceuticals, etc., and has safety in which no side effects are observed as a result of human clinical trials. The soybean lecithin has the characteristics of being insoluble or swollen in water, soluble in alcohol, ether, and chloroform, and insoluble in acetone. A generally known commercially available product is used, for example, the soybean lecithin may contain about 30% oil, and as phospholipids, may contain about 29% lecithin, about 31% cephalexin, and about 40% inositol-containing phospholipids.

[0027] According to one embodiment, the nonionic surfactant may be a biocompatible surfactant. According to one embodiment, the nonionic surfactant may be a liquid (or viscous liquid) nonionic surfactant. According to one embodiment, the nonionic surfactant may comprise a nonionic surfactant comprising a polyoxyethylene sorbitan ester (e.g., polysorbate (Tween trademark(®)). For example, the nonionic surfactant may be selected from the group consisting of Tween 20 (polyoxyethylene (20) sorbitan monolaurate, HLB 16.0), Tween 40 (polyoxyethylene (20) sorbitan monoopalmitate, HLB 15.6), Tween 60 (polyoxyethylene sorbitan monoostearate, HLB 14.9), Tween 65 (polyoxyethylene sorbitantristearate, HLB 10.5), Tween 80 (polyoxyethylene (20) sorbitan monooleate, HLB 15.0), and Tween 85 (polyoxyethylene sorbanetrioleate, HLB 11.0). It may include one or more types.

[0028] According to one embodiment, the nonionic surfactant may further include a “Cremophor® trademark nonionic surfactant comprising polyoxyethylated castor oil.” For example, it may be PolyoxyCremophor®REL (polyoxyethylated castor oil, HLB 2-14), Cremophor®RH40 (hydrogenated polyoxyethylated castor oil, HLB 14-16), Cremophor®RH60 (hydrogenated polyoxyethylated castor oil, HLB 15-17), Cremophor®RO (hydrogenated polyoxyethylated castor oil, HLB 16.1), etc. The aforementioned nonionic surfactants are commercially available and can be purchased from various suppliers. The Cremophor® trademark nonionic surfactant may be included in an amount of 1% to 20% by weight of the nonionic surfactant.

[0029] According to one embodiment, the nonionic surfactant may be applied as a single nonionic surfactant; or a nonionic surfactant complex composed of two or more nonionic surfactants.

[0030] According to one embodiment, the nonionic surfactant complex may be distinguished into a first nonionic surfactant and the remaining nonionic surfactants excluding the first nonionic surfactant (hereinafter referred to as surfactant A) (e.g., second surfactant, third surfactant, etc.), and these may be classified according to their HLB values. That is, the first nonionic surfactant may have an HLB greater than 15, i.e., an HLB of 16 or higher, and surfactant A may be a nonionic surfactant with an HLB of 15 or lower. For example, the first surfactant is Tween 20 (polyoxyethylene (20) sorbitan monolaurate, HLB 16.0), and the surfactant A may include one or more selected from the group consisting of Tween 60 (polyoxyethylene sorbitan monoostearate, HLB 14.9), Tween 65 (polyoxyethylene sorbitantristearate, HLB 10.5), Tween 80 (polyoxyethylene (20) sorbitan monooleate, HLB 15.0) and Tween 85 (polyoxyethylene sorbanetrioleate, HLB 11.0). For example, the first nonionic surfactant may be Tween 20 (polyoxyethylene (20) sorbitan monolaurate, HLB 16.0) and the second surfactant may be Tween 80 (polyoxyethylene (20) sorbitan monooleate, HLB 15.0). Here, the HLB value corresponds to the hydrophilic-lipophilic balance value.

[0031] According to one embodiment, the mixing ratio (based on mass) of the first nonionic surfactant to the surfactant A in the nonionic surfactant complex may be 3:1 to 2; or 3:1 to 1.5. By applying the above mixing ratio, a stable and efficient liposomal reaction can be carried out.

[0032] According to one embodiment, the nonionic surfactant complex may be prepared by stirring two or more nonionic surfactants in a solvent-free environment at a temperature of 20°C to 25°C for 5 to 10 minutes.

[0033] According to one embodiment, the second step (200), which involves adding and mixing a liposomal target substance to a phospholipid dispersion and an oil phase, may involve adding the liposomal target substance to the phospholipid dispersion and an oil phase at a specific ratio and mixing using a stirrer at 50 rpm to 100 rpm for 5 to 10 minutes to form a reaction mixture. Additionally, if necessary, solvents and additives may be further included, for example, purified water and reducing sugars (e.g., dextrin) may be added and mixed to complete the pre-liposomal step. For example, the mixing ratio (based on weight) of the liposomal target substance to the purified water may be 1:1.5 to 6. By applying the above mixing ratio, a stable and efficient liposomal reaction can be carried out, and productivity can be improved.

[0034] According to one embodiment, the additive may be a reducing sugar (e.g., dextrin) that can help with binding strength by having reactivity with other compounds during the liposomal formation process, and the mixing ratio (based on weight) of the nonionic surfactant to the additive (e.g., reducing sugar) may be 1:1 to 1:2.

[0035] According to one embodiment, the liposomal target substance is a raw material (or target) to be liposomalized and may be a health functional food ingredient. The liposomal target substance may be in a liquid, oil, or solid form (e.g., powder). According to one embodiment, the health functional food ingredient may be a bioactive substance. According to one embodiment, the health functional food ingredient may be an animal or plant extract; an animal or plant concentrate; or a chemical species or compound having a bioactive function (e.g., vitamin). For example, the health functional food ingredient may be red ginseng concentrate, red ginseng extract, vitamin C or a derivative thereof, maca extract, etc. Such health functional food ingredients are commercially available or obtained by extraction, concentration, synthesis, etc., through commonly known methods.

[0036] According to one embodiment, the mixing ratio (based on weight; w / w) of the liposomal target material to the soybean lecithin may be 1:1 to 3; or 1:1 to 2.5. This is appropriately adjusted for stable liposomal formation and can improve liposomal productivity.

[0037] According to one embodiment, the reaction mixture corresponds to a raw material mixture for a liposomal reaction, and the composition ratio (i.e., composition ratio) of the mixture can be determined by adjusting the mixing ratio according to the liposomal target material. Such composition ratio can calculate standard process conditions for process standardization and industrial mass production according to the liposomal target material. In addition, the efficiency and productivity of the liposomal reaction can be improved depending on the liposomal target material.

[0038] For example, among the above reaction mixture A:

[0039] It may include 24% to 40% by weight of soybean lecithin; 1% to 3% by weight of nonionic surfactant; 10% to 25% by weight of liposomal target material; and the remainder or 35% to 64% by weight of purified water.

[0040] For example, among the above reaction mixture B:

[0041] It may comprise 24% to 40% by weight of soybean lecithin; 1% to 3% by weight of nonionic surfactant; 10% to 25% by weight of liposomal target material; the remainder or 35% to 64% by weight of purified water; and 1% to 3% by weight of additive (reducing sugar).

[0042] For example, depending on the liposomal target material in each of the above reaction mixture A and the above reaction mixture B, the composition ratio may be as follows:

[0043] (1) Liposomal target substance: Vitamin C

[0044] 23% to 25% by weight soybean lecithin,

[0045] 1.2 wt% to 1.5 wt% nonionic surfactant

[0046] 23 to 26 weight% Vitamin C

[0047] 47% to 49% by weight of purified water, and

[0048] It may contain 2.3% to 2.5% by weight of reducing sugar.

[0049] (2) Liposomal target substance: Red ginseng concentrate

[0050] 35% to 37% by weight soybean lecithin,

[0051] 1.8 wt% to 2 wt% nonionic surfactant,

[0052] 24% to 26% by weight red ginseng concentrate,

[0053] and may include 35% to 37% by weight of purified water.

[0054] (3) Liposomal target substance: Maca extract

[0055] 24% to 26% by weight soybean lecithin,

[0056] 1.2 wt% to 1.5 wt% nonionic surfactant,

[0057] 10% to 12% by weight of maca extract,

[0058] 60% to 62% by weight of purified water and

[0059] It may contain 1.2% to 1.4% by weight of reducing sugar.

[0060] According to one embodiment, the third step (300), which sets the stirring speed, temperature, and time and proceeds with the liposomal reaction, can adjust the stirring speed, temperature, and time for the final step of liposomalization using the reaction mixture obtained in the second step (200). Depending on the characteristics of the raw material to be liposomed (i.e., the liposomal target material), the mixing ratio of the phospholipid and other components, the stirring speed, and the reaction temperature can be adjusted.

[0061] According to one embodiment, the third step (300) may produce liposomal formation by performing a three-stage stirring process in which at least one of the stirring time, stirring speed, or stirring time, or a combination thereof, is varied or continuously increased. For example, the three-stage stirring process may include: a first stirring step of stirring at 300 rpm to 500 rpm for 0 to 5 minutes; or 1 to 5 minutes; a second stirring step of stirring at 1,300 rpm to 1,500 rpm for 5 to 15 minutes; and a third stirring step of stirring at 1,800 rpm to 2,000 rpm for 15 to 30 minutes.

[0062] According to one embodiment, the temperature of the first stirring step may be 20 ℃ to 25 ℃, the temperature of the second stirring step may be 30 ℃ to 40 ℃, and the temperature of the third stirring step may be 40 ℃ to 50 ℃.

[0063] According to one embodiment, data such as the degree of liposomal formation, particle size, structure, and stabilization are collected based on the type of liposomal target material, the mixing ratio of components, the input amount, the stirring speed, the stirring temperature, and the stirring time. The collected data is then processed and converted into information to secure standard process conditions. This enables the standardization of liposomal manufacturing methods for each raw material group. For example, Vitamin C (crystalline powder), red ginseng concentrate (oil), and maca extract (powder) can be classified as standard liposomal target materials, and their optimal process conditions can be secured. The secured process conditions can be utilized as standard conditions.

[0064] For example, new target materials for liposomalization can be selected and identified, matched with standard target materials classified by raw material group, and relevant standard process conditions selected, followed by the liposomalization process. This can reduce the liposomalization process steps and time, and improve the efficiency of the process. For instance, standard conditions for liposomalization manufacturing methods by raw material group can be utilized in industrial-scale liposome production systems, enabling scale-up from laboratory scale to industrial production scale and improving the productivity and quality of liposomes.

[0065] According to one embodiment, the liposomal manufacturing method of the present invention can produce liposomal particles having a particle size ranging from a nanoscale of 10 nm or more to a scale of thousands of micrometers by controlling process conditions. The size of the liposomal particles in the present invention is 300 nm (nanometer) to 2,000 nm (nanometer).

[0066] According to one embodiment, the size and particle size distribution of liposome particles can be measured in a commonly known manner using dynamic light scattering measurement (i.e., Photon Correlation Spectroscopy (PCS) or Quasi-Elastic Light Scattering (QELS)) or laser diffraction measurement. The morphology and structure of liposomes can be analyzed using imaging techniques such as transmission electron microscopy (TEM), cryoelectron microscopy, atomic force microscopy (AFM), and small angle X-ray scattering (SAXS).

[0067] The embodiments of the present invention will be described in more detail below. However, the following embodiments are provided merely to aid in understanding the present invention and do not limit the scope of the present invention.

[0068] <Example 1>

[0069] Example regarding the manufacture of liposomal vitamin C.

[0070] Table 1 shows the composition ratio of input raw materials for the production of liposomal vitamin C.

[0071] item ratio Lecithin: Tween Complex 20 weight : 1 weight Tween Complex [Tween 20 : Tween 80] 3 weight : 1 weight Vitamin C: Lecithin 1 weight : 1 weight Purified water: Vitamin C 2 weight : 1 weight Tween complex: reducing sugar 1 weight : 2 weight

[0072] As shown in Table 1, each ingredient was standardized to a certain ratio and used in the experiment to manufacture liposomal vitamin C.

[0073] Table 2 shows the percentage of raw materials used in the production of liposomal vitamin C.

[0074] To form phospholipids, soybean lecithin and a Tween complex were added to a mixing container and mixed for 5 to 10 minutes at a stirring speed of 50 to 100 rpm to prepare a phospholipid dispersion and an oil phase. Vitamin C was added to the phospholipid dispersion and oil phase in a certain ratio and thoroughly mixed using a stirrer at 50 to 100 rpm for 5 to 10 minutes. During mixing, a certain ratio of purified water and reducing sugar (sucrose) was added and mixed to complete the pre-liposomal stage. For the final stage of liposomalization, the stirrer speed was adjusted from 500 rpm to 2,000 rpm and the temperature from 25 ℃ to 50 ℃. Liposomalization was performed by adjusting the stirring speed and temperature for each stage: the first stage for 0 to 5 minutes (speed 500 rpm and temperature 25 ℃), the second stage for 5 to 15 minutes (speed 1,500 rpm and temperature 40 ℃), and the third stage for 15 to 30 minutes (speed 2,000 rpm and temperature 50 ℃).

[0075] item Input Amount [Percentage (%)] lecithin 1 kg [24.1 %] Tween complex Tween 20 37.5 g [0.9 %] Tween 80 12.5 g [0.3 %] Vitamin C 1 kg [24.1 %] purified water 2 kg [48.2 %] reducing sugar 100 g [2.4%] Total 4.150 kg [100 %]

[0076] <Example 2>

[0077] Example regarding the preparation of liposomal red ginseng concentrate.

[0078] Table 3 shows the composition ratio of input raw materials for the production of liposomal red ginseng concentrate.

[0079] item ratio Lecithin: Tween Complex 20 weight : 1 weight Tween Complex [Tween 20 : Tween 80] 3 weight : 1 weight Red Ginseng Concentrate: Lecithin 1 weight : 1.5 weight Purified water: Red ginseng concentrate 1.5 weight : 1 weight

[0080] As shown in Table 3, each ingredient was standardized to a certain ratio and used in the experiment to manufacture liposomal red ginseng concentrate.

[0081] Table 4 shows the percentage of raw materials used in the production of liposomal red ginseng concentrate.

[0082] item Input Amount [Percentage (%)] lecithin 1.5 kg [36.8 %] Tween complex Tween 20 55 g [1.4 %] Tween 80 20 g [0.5 %] Red ginseng concentrate 1 kg [24.5 %] purified water 1.5 kg [36.8 %] Total 4.075 kg [100 %]

[0083] To form phospholipids, soybean lecithin and a Tween complex were added to a mixing container and mixed at a stirring speed of 50 to 100 rpm for 5 to 10 minutes to prepare a phospholipid dispersion and an oil phase. Red ginseng concentrate was added to the phospholipid dispersion and oil phase in a certain ratio and thoroughly mixed using a stirrer at 50 to 100 rpm for 5 to 10 minutes. During mixing, a certain ratio of purified water was added to complete the pre-liposome formation step.

[0084] For the final stage of liposomalization, the stirring speed was adjusted from 500 rpm to 2,000 rpm and the temperature from 25 ℃ to 50 ℃. Liposomalization was performed by adjusting the stirring speed and temperature for each stage: the first stage for 0 to 5 minutes (speed 500 rpm and temperature 25 ℃), the second stage for 5 to 15 minutes (speed 1,500 rpm and temperature 40 ℃), and the third stage for 15 to 30 minutes (speed 2,000 rpm and temperature 50 ℃).

[0085] <Example 3>

[0086] Example regarding the preparation of liposomal maca extract.

[0087] Table 5 shows the composition ratio of input raw materials for the production of liposomal maca extract.

[0088] item ratio Lecithin: Tween Complex 20 weight : 1 weight Tween Complex [Tween 20 : Tween 80] 3 weight : 1 weight Maca Extract: Lecithin 1 weight : 2.5 weight Purified water: Maca extract 6 weight : 1 weight Tween complex: reducing sugar 1 weight : 1 weight

[0089] As shown in Table 5, each raw material was standardized to a certain ratio and used in the experiment to prepare liposomal maca extract.

[0090] Table 6 shows the percentage of raw materials used in the production of liposomal maca extract.

[0091] item Input Amount [Percentage (%)] lecithin 2.5 kg [25.6 %] Tween complex Tween 20 93.8 g [1.0 %] Tween 80 31.2 g [0.3 %] Maca extract 1 kg [10.3 %] purified water 6 kg [61.5 %] reducing sugar 125 g [1.3 %] Total 9.750 kg [100 %]

[0092] To form phospholipids, soybean lecithin and a Tween complex were added to a mixing container and mixed for 5 to 10 minutes at a stirring speed of 50 to 100 rpm to prepare a phospholipid dispersion and an oil phase. Maca extract (powder) was added to the phospholipid dispersion and oil phase in a certain ratio and thoroughly mixed using a stirrer at 50 to 100 rpm for 5 to 10 minutes. During mixing, a certain ratio of purified water was added to complete the pre-liposomal stage. For the final stage of liposomalization, the stirrer speed was adjusted from 500 rpm to 2,000 rpm and the temperature from 25 ℃ to 50 ℃. Liposomalization was performed by adjusting the stirring speed and temperature for each stage: the first stage for 0 to 5 minutes (speed 500 rpm and temperature 25°C), the second stage for 5 to 15 minutes (speed 1,500 rpm and temperature 40°C), and the third stage for 15 to 30 minutes (speed 2,000 rpm and temperature 50°C).

[0093] The particle size and shape of the liposomal materials prepared in Examples 1 to 3 were measured using a hematocytometer slide under an optical microscope, which allows the manufacturer to easily observe whether liposomal formation has occurred.

[0094] It was confirmed that spherical liposome particles with a size of 300 nm to 1,500 nm were obtained in Example 1, spherical liposome particles with a size of 500 nm to 2,000 nm were obtained in Example 2, and spherical liposome particles with a size of 1,000 nm to 2,000 nm were obtained in Example 3. It was observed that the size of the liposomal particles of the red ginseng concentrate in Example 2 and the maca extract in Example 3, which had lower particle uniformity than that of Vitamin C in Example 1, was larger.

[0095] According to one embodiment, the present invention can provide a liposomalization process for functional food ingredients by forming an oil phase without using an organic solvent and setting stepwise stirring speeds and treatment temperatures in the physical treatment for liposomalization.

[0096] In addition, the present invention can establish categories of liposomal manufacturing methods by raw material group to improve the conventional liposome manufacturing process and to resolve the difficulty of standardizing liposomal manufacturing methods, as the manufacturing methods for each functional food ingredient to be liposomalized vary significantly.

[0097] Furthermore, the present invention provides a liposomal manufacturing method that facilitates industrial-scale production by standardizing the entire process, and can provide benefits to overall management costs by shortening the liposomal manufacturing process and time.

[0098] Furthermore, the present invention provides a liposomal manufacturing method that facilitates industrial-scale production and can improve product performance by increasing the absorption rate of health functional food ingredients. Additionally, by ensuring established quality consistency and applying it to various ingredients, it can provide industrial benefits that lead to product diversity.

[0099] The foregoing description is merely an illustrative explanation of the technical concept of the present invention, and those skilled in the art to which the present invention pertains will be able to make various modifications and variations within the scope of the essential characteristics of the present invention. Accordingly, the embodiments disclosed in the present invention are intended to explain, not to limit, the technical concept of the present invention, and the scope of the technical concept of the present invention is not limited by such embodiments. The scope of protection of the present invention shall be interpreted by the claims, and all technical concepts within an equivalent scope shall be interpreted as being included within the scope of rights of the present invention.

Claims

Claim 1 A first step of preparing a phospholipid dispersion and an oil phase forming product by mixing a nonionic surfactant and soybean lecithin; a second step of forming a reaction mixture by adding a liposomal target substance to the phospholipid dispersion and oil phase forming product and mixing. A third step comprising carrying out a liposomal reaction in the reaction mixture using a three-stage stirring process in which the stirring time, stirring speed, and temperature are continuously increased; wherein the three-stage stirring process of the third step comprises: a first stirring step of stirring at 100 rpm to 500 rpm for 0 to 5 minutes; a second stirring step of stirring at 600 rpm to 1,500 rpm for 5 to 15 minutes; and a third stirring step of stirring at 1,600 rpm to 2,000 rpm for 15 to 30 minutes; wherein the temperature of the second stirring step is 5 ℃ to 20 ℃ higher than the temperature of the first stirring step, and the temperature of the third stirring step is 5 ℃ to 10 ℃ higher than the temperature of the second stirring step, and the mixing ratio (w / w) of the soybean lecithin to the nonionic surfactant is A liposomal manufacturing method in which the ratio is 20:1 to 5, and the first step of preparing the phospholipid dispersion and oil phase forming product is solvent-free. Claim 2 A method for manufacturing liposomal formation according to claim 1, wherein the nonionic surfactant is introduced as a nonionic surfactant complex composed of a single nonionic surfactant or two or more types of nonionic surfactants, and the nonionic surfactant is a liquid nonionic surfactant comprising polyoxyethylene sorbitan ester, and the mixing ratio (w / w) of the first nonionic surfactant to the remaining surfactant in the nonionic surfactant complex is 3:1 to 2. Claim 3 A method for manufacturing liposomal formation according to claim 2, wherein the first nonionic surfactant among the nonionic surfactant complex is Tween 20, and the remaining nonionic surfactants comprise one or more selected from the group consisting of Tween 40, Tween 60, Tween 65, Tween 80, and Tween 85. Claim 4 A method for manufacturing liposomalization according to claim 1, wherein the liposomalization target substance is a functional food ingredient, and the functional food ingredient is an animal or plant extract; an animal or plant concentrate; or a vitamin. Claim 5 A liposomal manufacturing method according to claim 4, wherein the functional food ingredient is one or more selected from the group consisting of red ginseng concentrate, maca extract, and vitamin C. Claim 6 A method for manufacturing liposomalization according to claim 1, wherein the mixing ratio (based on weight) of the liposomalization target substance to the soybean lecithin is 1:1 to 3. Claim 7 A method for manufacturing liposomalization according to claim 1, wherein the liposomalization target substance is added together with purified water, and the mixing ratio (w / w) of the liposomalization target substance to the purified water is 1:1.5 to 6. Claim 8 A method for producing liposomalization according to claim 1, comprising 24% to 40% by weight of soybean lecithin, 1% to 3% by weight of a nonionic surfactant, 10% to 25% by weight of a liposomal target substance, and 35% to 64% by weight of purified water in the reaction mixture, and further comprising 1% to 3% by weight of a reducing sugar. Claim 9 A method for manufacturing liposomal formation according to claim 1, wherein the first step is stirring for 5 to 10 minutes at a stirring speed of 50 rpm to about 200 rpm. Claim 10 delete Claim 11 delete Claim 12 In claim 1, the liposomal manufacturing method is a method of providing liposomal particles with a size of 300 nm (nanometer) to 2,000 nm (nanometer).

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