Encapsulation product of aqueous-phase NAD < + > precursor and encapsulation method and application thereof

Abstract

The invention belongs to the technical field of packaging, and particularly relates to a packaging product of a water-phase NAD + precursor and a packaging method and application of the packaging product. Comprising the following steps: preparing a water-phase solution containing an NAD + precursor; preparing a packaging solution containing an oil phase material and a surfactant; and preparing a micro-fluidic chip, and preparing the packaging solution and the water-phase solution into a water-in-oil core-shell structure by adopting the micro-fluidic chip through a micro-fluidic technology, so as to obtain a packaging product of the water-phase NAD + precursor. The packaging method is easy to operate and high in efficiency, the prepared packaging product is of a water-in-oil structure, the water-phase NAD + precursor is packaged into an oil-phase shell layer, interference of factors such as oxygen, ultraviolet light, temperature and pH in the environment on the stability of the water-phase NAD + precursor is isolated, the stability and biological activity of the water-phase NAD + precursor in the storage, transportation and application process are remarkably improved, and the packaging product is suitable for industrial production. The application potential in the fields of skin care and health care products is widened.

Description

Technical Field

[0001] The present invention relates to a packaging technology field, and more particularly to a water-phase NAD + Precursor encapsulation product and encapsulation method and application thereof. Background Art

[0002] Nicotinamide adenine dinucleotide (NAD + ) is an indispensable coenzyme in cell metabolism, and is involved in a series of key biochemical reactions, including but not limited to: activating the Sirtuin protein family for protein deacetylation, promoting the repair of DNA damage, maintaining antioxidant balance, regulating inflammatory responses and energy metabolism pathways, etc. These mechanisms together constitute NAD + It plays a fundamental role in maintaining cell function, delaying biological aging, enhancing immunity, improving cognitive ability, and protecting the liver and vision. + The natural decline in NAD levels is considered an important biomarker of the aging process, which has triggered scientific research on how to effectively maintain or increase NAD levels. + level of widespread attention.

[0003] Direct NAD supplementation + It may cause it to be rapidly metabolized and excreted in the body, thereby increasing NAD in cells + The strategy to increase NAD levels has turned to supplementing its precursors. Currently, compounds such as nicotinamide mononucleotide (NMN), nicotinamide riboside (NR), and reduced nicotinamide adenine dinucleotide (NADH, or NAD(P)H) are considered to be the most effective way to increase NAD levels. + In particular, NADH can be quickly converted into NAD + , and in this conversion process, protons (H + ) and adenosine triphosphate (ATP), of which H + It helps to remove free radicals that may cause cellular aging, while ATP provides support for the energy needs of cells. Therefore, NADH supplementation is regarded as a highly effective strategy to enhance intracellular NAD + level.

[0004] However, NADH and NAD + The precursor is highly sensitive to environmental factors such as ultraviolet light, temperature, oxygen and pH, and there are problems of instability and inactivation during production, storage and transportation due to these environmental factors. Although NADH products in the form of freeze-dried powder have a longer shelf life than those in solution form, this does not fundamentally solve its stability problem, which seriously limits its application. Summary of the invention

[0005] The purpose of this application is to provide an aqueous NAD + The encapsulation product of the precursor and its encapsulation method and application are intended to solve the problem of NADH and other NAD + The precursor is highly sensitive to environmental factors such as ultraviolet light, temperature, oxygen and pH. Aqueous precursors are affected by these environmental factors and have unstable performance and are easily inactivated during production, storage and transportation.

[0006] In order to achieve the above application purpose, the technical solution adopted in this application is as follows:

[0007] In the first aspect, the present application provides an aqueous phase NAD + The encapsulation method of the precursor comprises the following steps:

[0008] Preparation containing NAD + an aqueous solution of a precursor;

[0009] preparing an encapsulation solution comprising an oil phase material and a surfactant;

[0010] Prepare a microfluidic chip, use the microfluidic chip to make the encapsulation solution and the aqueous solution into an oil-in-water core-shell structure through microfluidic technology, and obtain aqueous NAD + Encapsulation product of the precursor.

[0011] In some possible implementations, the NAD + The precursor includes at least one of NAD(P)H, NMN, and NR.

[0012] In some possible implementations, the oil phase material includes at least one of mineral oil, vegetable oil, silicone oil, fluorinated oil, and synthetic oil.

[0013] In some possible implementations, the surfactant includes at least one of Span 80, Tween 20, sodium lauryl sulfate, sodium lauryl ether sulfate, cetyltrimethylammonium chloride, polyoxyethylene lauryl alcohol, polyoxyethylene phosphate, olive oil soap, and glucoside.

[0014] In some possible implementations, the aqueous solution further includes an antioxidant and a buffer solution.

[0015] In some possible implementations, the encapsulation solution further includes a polymer carrier or a polymer carrier and a curing agent.

[0016] In some possible implementations, the NAD + The precursor is selected from NAD(P)H.

[0017] In some possible implementations, in the aqueous phase solution, the concentration of the buffer solution is 0.01 mol / L to 1 mol / L, the pH value is 7.0 to 7.5, and the NAD + The concentration of the precursor is 0.1 mmol / L to 10 mmol / L, and the mass percentage of the antioxidant is 1% to 20%.

[0018] In some possible implementations, the antioxidant includes at least one of vitamin C, vitamin E, anthocyanin, chlorophyll, carotene, and glucose.

[0019] In some possible implementations, the buffer solution includes at least one of a phosphate buffered saline solution and a Tris buffer solution.

[0020] In some possible implementations, in the encapsulation solution, the mass percentage of the oil phase material is 80% to 95%, and the mass percentage of the surfactant is 0.02% to 5%.

[0021] In some possible implementations, in the encapsulation solution, the added amount of the polymer carrier is 0.05% to 10% of the mass of the oil phase material.

[0022] In some possible implementations, in the encapsulation solution, the added amount of the curing agent is 0.05% to 10% of the mass of the oil phase material.

[0023] In some possible implementations, the polymer carrier includes at least one of alginate, polysaccharide, polylactic acid-glycolic acid copolymer, and protein.

[0024] In some possible implementations, the curing agent includes at least one of epoxy resin, polyamine curing agent, acrylate monomer, 2-hydroxy-2-methylacetone, protein, polysaccharide, glutaraldehyde, sodium alginate, and calcium ions.

[0025] In some possible implementations, the method for preparing the microfluidic chip includes: at least one of photolithography technology and 3D printing technology.

[0026] In some possible implementations, the microfluidic chip includes an inlet area for an aqueous solution, an inlet area for an encapsulation solution, an intersection, and an outlet area, wherein the inlet area for the aqueous solution is connected to the intersection through an input flow channel for the aqueous solution, the inlet area for the encapsulation solution is connected to the intersection through an input flow channel for the encapsulation solution, and the intersection is connected to the outlet area through a droplet flow channel.

[0027] In some possible implementations, the steps of the microfluidic technology include: transporting the aqueous phase solution and the encapsulation solution from the inlet region to the intersection through the input flow channel, generating oil-in-water droplets at the intersection through the microfluidic technology, and the oil-in-water droplets are collected in the outlet region through the droplet flow channel to obtain the aqueous phase NAD + Encapsulation product of the precursor.

[0028] In some possible implementations, the method of generating water-in-oil droplets by microfluidics technology includes one of T-type structure, flow focusing, cross flow, coaxial flow, and step emulsification;

[0029] In some possible implementations, the ratio of the flow rates of the aqueous solution and the encapsulation solution reaching the intersection is (4-7): (10-13);

[0030] In some possible implementations, the flow rate of the aqueous solution reaching the intersection is 20 μL / h to 35 μL / h.

[0031] In some possible implementations, the flow rate of the encapsulation solution reaching the intersection is 50 μL / h to 65 μL / h.

[0032] In some possible implementations, in the microfluidic chip, a buffer zone is further provided between the droplet flow channel and the outlet area.

[0033] In some possible implementations, the inner diameter of the inlet region of the aqueous solution is 800 μm to 1200 μm.

[0034] In some possible implementations, the inner diameter of the inlet region of the encapsulated solution is 800 μm to 1200 μm.

[0035] In some possible implementations, the inner diameter of the input flow channel of the aqueous solution is 10 μm to 500 μm.

[0036] In some possible implementations, the inner diameter of the input channel of the encapsulation solution is 10 μm to 500 μm.

[0037] In some possible implementations, the inner diameter of the droplet flow channel is 10 μm to 500 μm.

[0038] In some possible implementations, the inner diameter of the flow channel of the buffer zone is 10 μm to 500 μm.

[0039] In some possible implementations, the ratio of the length of the flow channel of the buffer zone to the length of the droplet flow channel is (2-10):1.

[0040] In some possible implementations, in the input flow channel of the aqueous solution, the inner diameter of the end close to the intersection is smaller than the inner diameter of the end close to the inlet region of the aqueous solution.

[0041] In some possible implementations, in the input flow channel of the encapsulated solution, the inner diameter of the end close to the intersection is smaller than the inner diameter of the end close to the inlet region of the encapsulated solution.

[0042] In some possible implementations, in the input flow channel of the aqueous solution, the ratio of the inner diameter at one end close to the intersection to the inner diameter at one end close to the inlet region of the aqueous solution is (1.5-2.5):1.

[0043] In some possible implementations, in the input flow channel of the encapsulation solution, the ratio of the inner diameter of the end close to the intersection to the inner diameter of the end close to the inlet area of ​​the encapsulation solution is (1.5-2.5):1.

[0044] In some possible implementations, the ratio of the inner diameter of the input flow channel of the aqueous solution to the inner diameter of the input flow channel of the encapsulation solution is 1:1.

[0045] In a second aspect, the present invention provides an aqueous NAD + The encapsulation product of the precursor, the aqueous phase NAD + The encapsulated product of the precursor is an oil-in-water structure, including NAD + The precursor has a water phase inner core and an oil phase encapsulation layer, wherein the oil phase encapsulation layer comprises an oil phase material and a surfactant.

[0046] In some possible implementations, the aqueous phase NAD + The average particle size of the encapsulated product of the precursor is 50 μm to 100 μm.

[0047] In some possible implementations, the aqueous phase NAD + In the encapsulated product of the precursor, the mass ratio of the water phase inner core to the oil phase encapsulation layer is (20-35):(50-65).

[0048] In some possible implementations, the aqueous core further includes an antioxidant and a buffer solution.

[0049] In some possible implementations, the outer surface of the oil phase encapsulation layer further includes a polymer carrier layer, and the thickness of the polymer carrier layer is 50 nm to 200 nm.

[0050] In some possible implementations, the aqueous phase NAD + The encapsulated product of the precursor is stored in a sealed, light-proof environment at a temperature of -20°C to 25°C.

[0051] In a third aspect, the present invention provides an aqueous NAD + The encapsulation product of the precursor is used to prepare the aqueous NAD + The encapsulated product of the precursor or the aqueous NAD + The encapsulated product of the precursor is applied to at least one of the fields of skin care products, health care products and medicines.

[0052] The aqueous NAD provided in the first aspect of the present application + Precursor encapsulation method, respectively preparing NAD + The aqueous phase solution of the precursor and the encapsulation solution containing the oil phase material and the surfactant, wherein the surfactant acts on the interface between the aqueous phase and the oil phase to promote the formation of the oil-in-water encapsulation product, improve the stability of the encapsulation product, and reduce the risk of agglomeration of the encapsulation product. Prepare a microfluidic chip, use the microfluidic chip through microfluidic technology, make the encapsulation solution and the aqueous phase solution into an oil-in-water core-shell structure, and realize the aqueous phase NAD + Precise control of the droplet size, uniformity and encapsulation efficiency of the precursor oil-in-water encapsulation results in the acquisition of aqueous NAD + The encapsulation method of the present application is simple to operate and highly efficient. The encapsulated product obtained is an oil-in-water structure, and the aqueous phase NAD + The precursor is encapsulated in the oil phase shell layer, isolating the environmental factors such as oxygen, ultraviolet light, temperature and pH from affecting the aqueous phase NAD + The interference of precursor stability significantly increased the aqueous phase NAD + The stability and biological activity of the precursor during storage, transportation and application broaden its application potential in the field of skin care and health products.

[0053] This application aqueous NAD + The encapsulated product of the precursor is an oil-in-water structure, including NAD + The precursor has an aqueous core and an oil phase encapsulation layer, wherein the oil phase encapsulation layer comprises an oil phase material and a surfactant. The coating of the oil phase encapsulation layer significantly increases the NAD + Stability of precursors, effective protection of NAD + The precursor is not affected by factors such as oxygen, ultraviolet light, temperature and pH in the environment, avoiding the presence of aqueous NAD + Precursor inactivation significantly increases aqueous NAD + Stability of precursors during storage, transportation and application.

[0054] This application aqueous NAD + The encapsulation product of the precursor contains NAD stably +Precursors have the functions of maintaining cell function, delaying biological aging, enhancing immunity, improving cognitive ability, protecting the liver and vision, and removing free radicals that may cause cell aging. They can not only be used to develop skin care products with anti-aging effects, but also can be used in health products and medicines with the effects of reverse aging, enhancing vitality, enhancing immunity, protecting the liver and vision, improving athletic ability and rapid recovery. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0056] Figure 1 It is the aqueous phase NAD provided in the examples of this application. + A schematic flow diagram of a precursor encapsulation method;

[0057] Figure 2 It is a schematic diagram of the structure of the microfluidic technology chip used in the encapsulation method of aqueous NADH provided in Example 1 of the present application;

[0058] Figure 3 It is a schematic diagram of the structure of the microfluidic technology chip used in the encapsulation method of aqueous NADH provided in Example 2 of the present application;

[0059] Figure 4 It is a schematic diagram of the structure of the microfluidic technology chip used in the encapsulation method of aqueous NADH provided in Example 3 of the present application;

[0060] Figure 5 It is a schematic diagram of droplet generation by coaxial flow, cross flow (T-shaped structure) and flow focusing in the microfluidic technology provided in the embodiments of the present application;

[0061] Figure 6 This is a physical picture of the droplet flow channel of the encapsulated product of aqueous NAD(P)H in the microfluidic technology chip provided in Example 2 of the present application;

[0062] Among them, the reference numerals in the figure are:

[0063] 1—Inlet area of ​​aqueous solution 2—Input flow channel of aqueous solution

[0064] 3—Inlet area of ​​encapsulation solution 4—Input flow channel of encapsulation solution

[0065] 5—intersection point 6—droplet flow channel 7—exit area 8—buffer zone. DETAILED DESCRIPTION

[0066] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application more clearly understood, the present application is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0067] In this application, the term "and / or" describes the association relationship of associated objects, indicating that there may be three relationships. For example, A and / or B can mean: A exists alone, A and B exist at the same time, and B exists alone. A and B can be singular or plural. The character " / " generally indicates that the associated objects are in an "or" relationship.

[0068] In this application, "at least one" means one or more, and "plurality" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single items or plural items. For example, "at least one of a, b or c", or "at least one of a, b and c", can all represent: a, b, c, ab (i.e. a and b), ac, bc, or abc, where a, b, c can be single or multiple, respectively.

[0069] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the above-mentioned processes does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0070] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments, and are not intended to limit the present application. The singular forms "a", "an" and "the" used in the embodiments of the present application and the appended claims are also intended to include plural forms, unless the context clearly indicates other meanings.

[0071] The weight of the relevant components mentioned in the embodiments of the present specification can not only refer to the specific content of each component, but also represent the proportional relationship between the weights of the components. Therefore, as long as the content of the relevant components in the embodiments of the present specification is proportionally enlarged or reduced, it is within the scope disclosed in the embodiments of the present specification. Specifically, the mass described in the embodiments of the present specification can be μg, mg, g, kg and other mass units known in the chemical industry.

[0072] The terms "first" and "second" are used only for descriptive purposes to distinguish objects such as substances from each other, and should not be understood as indicating or implying relative importance or implicitly indicating the number of indicated technical features. For example, without departing from the scope of the embodiments of the present application, the first XX may also be referred to as the second XX, and similarly, the second XX may also be referred to as the first XX. Thus, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features.

[0073] The term "microfluidics" refers to the science and technology involved in systems that use microchannels (with dimensions of tens to hundreds of microns) to process or manipulate tiny fluids (volumes of nanoliters to attoliters). It is an emerging interdisciplinary subject involving chemistry, fluid physics, microelectronics, new materials, biology, and biomedical engineering. Because of its miniaturization and integration, microfluidic devices are usually called microfluidic chips, also known as labs on a chip and micro-total analytical systems.

[0074] In a first aspect, an aqueous NAD + Precursor packaging methods, such as Figure 1 As shown, the following steps are included:

[0075] S10. Preparation of NAD + an aqueous solution of a precursor;

[0076] S20. Preparing an encapsulation solution comprising an oil phase material and a surfactant;

[0077] S30. Prepare a microfluidic chip, use the microfluidic chip to make the encapsulation solution and the aqueous solution into an oil-in-water core-shell structure through microfluidic technology, and obtain aqueous NAD + Encapsulation product of the precursor.

[0078] The aqueous NAD provided in the first aspect of the embodiment of the present application + Precursor encapsulation method, respectively preparing NAD + The aqueous solution of the precursor and the encapsulation solution comprising an oil phase material and a surfactant, wherein NAD + The precursor is soluble in the water phase and is compatible with the oil phase material that is insoluble in the encapsulation solution. The surfactant acts on the interface between the water phase and the oil phase to promote the formation of oil-in-water encapsulation products, improve the stability of the encapsulation products, and reduce the risk of encapsulation product agglomeration. Prepare microfluidic chips, use microfluidic chips and microfluidic technology to make the encapsulation solution and the water phase solution into an oil-in-water core-shell structure, and achieve the water phase NAD + Precise control of the droplet size, uniformity and encapsulation efficiency of the precursor oil-in-water encapsulation results in the acquisition of aqueous NAD+ The encapsulation method of the present application embodiment is simple to operate and highly efficient. The encapsulated product obtained is an oil-in-water structure, and the aqueous phase NAD + The precursor is encapsulated in the oil phase shell layer, isolating the environmental factors such as oxygen, ultraviolet light, temperature and pH from affecting the aqueous phase NAD + The interference of precursor stability significantly increased the aqueous phase NAD + The stability and biological activity of the precursor during storage, transportation and application broaden its application potential in the field of skin care and health products.

[0079] In the above step S10: preparing + Aqueous solution of the precursor.

[0080] In some possible implementations, NAD + Precursors include at least one of nicotinamide mononucleotide (NMN), nicotinamide riboside (NR), and reduced nicotinamide adenine dinucleotide (NADH, or NAD(P)H); these compounds are believed to increase NAD + Level of effective precursors.

[0081] In some possible implementations, NAD + The precursor is selected from NAD(P)H; NAD(P)H can not only be rapidly converted into NAD + In this conversion process, protons (H+) and adenosine triphosphate (ATP) can be generated and released, of which H+ helps to remove free radicals that may cause cell aging, while ATP supports the energy needs of cells. Therefore, the supplementation of NADH is regarded as an efficient strategy to enhance the intracellular NAD + Level of effective precursors.

[0082] In some possible implementations, the aqueous solution also contains an antioxidant and a buffer solution. In this case, by adding the antioxidant and the buffer solution, the oxidation reaction can be prevented and a suitable pH environment can be maintained, so that NAD + The precursor is not affected by oxidation and pH changes, thus protecting NAD in aqueous solutions + Activity of precursors.

[0083] In some possible implementations, in the aqueous phase solution, the concentration of the buffer solution is 0.01 mol / L to 1 mol / L, the pH value is 7.0 to 7.5, and NAD + The concentration of the precursor is 0.1mmol / L to 10mmol / L, and the mass percentage of the antioxidant is 1% to 20%. In this case, the aqueous phase solution uses water as the solvent, and the NAD +The precursor has better stability, is less affected by environmental oxidation and pH changes, and can better protect NAD in aqueous solutions + Activity of precursors. For example, in aqueous solution, NAD + The concentration of the precursor can be 0.1mmol / L, 1mmol / L, 2mmol / L, 3mmol / L, 4mmol / L, 5mmol / L, 6mmol / L, 7mmol / L, 8mmol / L, 9mmol / L, 10mmol / L or other typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values; the mass percentage of the antioxidant is 1%, 3%, 5%, 8%, 10%, 13%, 15%, 18%, 20% or other typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values; the concentration of the buffer solution can be 0.01mol / L, 0.03mol / L, 0.05mol / L, 0.08mol / L, 0.1mol / L or other typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values; the pH value is 7.0, 7.1, 7.2, 7.3, 7.4, 7.5 or the like.

[0084] In some possible implementations, the antioxidant includes at least one of vitamin C, vitamin E, anthocyanin, chlorophyll, carotene, and glucose; these antioxidants can prevent oxidation reactions and make NAD + The precursor is not affected by oxidation, thus protecting NAD in the aqueous solution + Activity of precursors.

[0085] In some possible implementations, the buffer solution includes at least one of a phosphate buffered saline solution and a tris buffer solution; these buffer solutions are conducive to maintaining the pH value of the aqueous phase solution between 7.0 and 7.5, reducing NAD + Precursors are affected by pH changes and protect NAD in aqueous solutions + Activity of precursors.

[0086] In the above step S20: an encapsulation solution comprising an oil phase material and a surfactant is prepared, wherein the oil phase can form a coating layer on the surface of the water phase droplet to form an oil-in-water structure, and the surfactant acts on the interface between the water phase and the oil phase to promote the formation of the oil-in-water encapsulation product, improve the stability of the encapsulation product, and reduce the risk of agglomeration of the encapsulation product.

[0087] In some possible implementations, the oil phase material includes at least one of mineral oil, vegetable oil, silicone oil, fluorinated oil, and synthetic oil; these oil phase materials can form an oil-in-water structure with the aqueous solution.

[0088] In some possible implementations, the surfactant includes at least one of Span 80, Tween 20, sodium lauryl sulfate, sodium lauryl ether sulfate, cetyltrimethylammonium chloride, polyoxyethylene lauryl alcohol, polyoxyethylene phosphate, olive oil soap, and glucoside; these surfactants can act on the interface between the water phase and the oil phase to promote the formation of oil-in-water encapsulation products, improve the stability of the encapsulation products, and reduce the risk of agglomeration of the encapsulation products.

[0089] In some possible implementations, in the encapsulation solution, the mass percentage of the oil phase material is 80% to 95%, and the mass percentage of the surfactant is 0.02% to 5%. In this case, the oil phase material is used as a solvent in the encapsulation solution, and the ratio of the oil phase material and the surfactant ensures that the encapsulation solution has sufficient surface tension, thereby ensuring that the encapsulation solution stably forms droplets in the microfluidic technology, and avoids the separation of the aqueous phase solution and the oil phase encapsulation solution to form a laminar flow. Exemplarily, in the encapsulation solution, the mass percentage of the oil phase material can be 80%, 85%, 90%, 95%, etc., which are typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values, and the mass percentage of the surfactant is 0.02%, 1%, 2%, 3%, 4%, 5%, etc., which are typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values.

[0090] In some possible implementations, the encapsulation solution also contains a polymer carrier or a polymer carrier and a curing agent. In this case, the oil phase in the encapsulation solution will first be coated on the surface of the water phase droplets. After the oil-in-water droplets are formed, the polymer carrier has a certain interfacial activity and will spontaneously migrate to the surface of the oil phase and adsorb on the outer surface of the oil phase coating layer to form a stable polymer carrier coating layer. The polymer carrier coating layer has a sustained release effect. Furthermore, when the polymer carrier and the curing agent are added at the same time, under the action of the curing agent, the polymer carrier layer solidifies into a solid shell layer, at which time the NAD in the water phase core is + The precursor can form a more stable protection and achieve multifunctional protection.

[0091] In some possible implementations, the amount of the polymer carrier added to the encapsulation solution is 0.05% to 10% of the mass of the oil phase material, and can be 0.05%, 1%, 2%, 3%, 4%, 5%, 6%, 8%, 10%, or any other typical but non-limiting point value or an interval value between any two points. In this case, the content of the polymer carrier can ensure that a polymer carrier layer is formed outside the encapsulation layer of the oil phase, and that the subsequent NAD in the water phase core is + Sustained release effect of precursors.

[0092] In some possible implementations, the polymer carrier includes at least one of alginate, polysaccharide, polylactic acid-glycolic acid copolymer, and protein; these polymer carrier materials can further form a polymer carrier layer on the surface of the encapsulation layer of the oil phase.

[0093] In some possible implementations, the amount of curing agent added to the encapsulation solution is 0.05% to 10% of the mass of the oil phase material, and specifically can be 0.05%, 1%, 2%, 3%, 4%, 5%, 6%, 8%, 10%, or any other typical but non-limiting point value or an interval value between any two point values. In this case, the concentration of the curing agent fully ensures the curing effect on the polymer carrier layer.

[0094] In some possible implementations, the curing agent includes at least one of epoxy resin, polyamine curing agent, acrylate monomer, 2-hydroxy-2-methyl acetone, protein, polysaccharide, glutaraldehyde, sodium alginate, and calcium ions. Among them, epoxy resin and polyamine curing agent are thermal curing agents, acrylate monomer and 2-hydroxy-2-methyl acetone are light curing agents, protein, polysaccharide, glutaraldehyde are chemical curing agents, and sodium alginate and calcium ions are ion cross-linking curing agents.

[0095] In the above step S30: prepare a microfluidic chip, use the microfluidic chip to process the encapsulation solution and the aqueous solution using microchannels (with a size of tens to hundreds of microns) through microfluidic technology to form an oil-in-water core-shell structure, and obtain aqueous NAD + Encapsulation product of the precursor.

[0096] In some possible implementations, the method for preparing the microfluidic chip includes: at least one of photolithography technology and 3D printing technology.

[0097] In some specific embodiments, the steps of preparing a microfluidic chip using photolithography technology include: coating a photoresist on a silicon wafer, and creating a desired microfluidic channel pattern by photolithography technology; then mixing polydimethylsiloxane (PDMS) and a curing agent in a ratio of 10:1, stirring evenly and pouring the mixture on a silicon wafer; curing at 80°C for 30 minutes; after curing, peeling the PDMS off the silicon wafer, and plasma bonding it to a glass sheet or another PDMS layer to form a complete microfluidic chip.

[0098] In some specific embodiments, the steps of preparing a microfluidic chip using 3D printing technology include: preparing a microfluidic channel pattern using 3D printing technology; then mixing polydimethylsiloxane (PDMS) and a curing agent in a ratio of 10:1, stirring evenly and pouring the mixture on a silicon wafer; curing at 80°C for 30 minutes; after curing, peeling the PDMS off the silicon wafer and plasma bonding it to a glass sheet or another PDMS layer to form a complete microfluidic chip.

[0099] In some possible implementations, such as the attached Figures 2 to 4 As shown, the microfluidic chip includes an inlet area 1 for an aqueous solution, an inlet area 3 for an encapsulation solution, an intersection 5 and an outlet area 7, wherein the inlet area 1 for the aqueous solution is connected to the intersection 5 through an input flow channel 2 for the aqueous solution, the inlet area 3 for the encapsulation solution is connected to the intersection 5 through an input flow channel 4 for the encapsulation solution, and the intersection 5 is connected to the outlet area 7 through a droplet flow channel 6.

[0100] In some possible implementations, the steps of microfluidic technology include: the steps of microfluidic technology include: transporting the aqueous phase solution and the encapsulation solution from the inlet area through the input flow channel to the intersection 5 respectively, generating oil-in-water droplets at the intersection 5 through microfluidic technology, and the oil-in-water droplets are collected in the outlet area 7 through the droplet flow channel 6 to obtain the encapsulation product of the aqueous phase NAD+ precursor.

[0101] In some possible implementations, the ratio of the flow rates of the aqueous solution and the encapsulation solution reaching the intersection 5 is (4-7): (10-13). In this case, the flow rate ratio of the aqueous solution and the oil-phase encapsulation solution fully ensures the effective contact and mixing of the two liquids to form stable "water-in-oil" droplets. Exemplarily, the ratio of the flow rates of the aqueous solution and the encapsulation solution reaching the intersection 5 can be 4:10, 5:11, 6:12, 7:13, etc., typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values.

[0102] In some possible implementations, the flow rate of the aqueous solution to the intersection 5 is 20 μL / h to 35 μL / h. For example, it can be 20 μL / h, 25 μL / h, 30 μL / h, 35 μL / h, and other typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values. In some possible implementations, the flow rate of the encapsulation solution to the intersection 5 is 50 μL / h to 65 μL / h. For example, it can be 50 μL / h, 55 μL / h, 60 μL / h, 65 μL / h, and other typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values. In the above embodiments of the present application, the flow rates of the aqueous solution and the oil-phase encapsulation solution ensure effective contact and mixing of the two liquids to form stable "water-in-oil" droplets.

[0103] In some possible implementations, the method of generating water-in-oil droplets by microfluidics technology includes one of T-type structure, flow focusing, cross flow, coaxial flow, and step emulsification. Figure 5As shown, the T-type structure refers to the intersection 5 where the aqueous solution and the oil-phase encapsulation solution contact and mix at the intersection of the T-type structure to form water-in-oil droplets. Flow focusing is a central main channel for the liquid phase (dispersed phase, i.e., aqueous solution) for droplet generation, and two side channels for carrying the continuous phase, i.e., the oil-phase encapsulation solution. These channels intersect at a confluence point, and the oil-phase encapsulation solution squeezes the aqueous solution from both sides, forcing it to pass through a narrow nozzle or orifice to form droplets. The cross type, also known as the T-type, is a main channel and a side channel perpendicular to it. The aqueous solution flows in from the main channel, while the oil-phase encapsulation solution flows in from the side channel and meets at the intersection, where the flow of the oil-phase encapsulation solution shears the aqueous solution, thereby forming droplets. The coaxial flow droplet generation method utilizes coaxially arranged channels, in which the inner channel carries the aqueous solution and the outer channel carries the oil-phase encapsulation solution. The aqueous solution flows in the inner channel, the oil-phase encapsulation solution flows in the outer channel, the two phases meet at the coaxial nozzle, and the oil-phase encapsulation solution surrounds the aqueous solution and forms droplets. Step emulsification is a method that uses geometric structure changes to generate droplets. This method usually involves a channel with a step-like structure, and the aqueous phase solution is sheared to form droplets due to the sudden change in geometric structure when flowing through the steps.

[0104] In some possible implementations, a buffer zone 8 is further provided between the droplet flow channel 6 and the outlet region 7 in the microfluidic chip. Figure 2 As shown, the chip of the microfluidic technology of the embodiment of the present application includes multiple flow channels, the aqueous solution is stored in the inlet area 1 of the aqueous solution, and is transported to the intersection 5 through the input flow channel 2 of the aqueous solution. At the same time, the encapsulation solution is stored in the inlet area 3 of the oil solution, and is transported to the intersection 5 through the input flow channel 4 of the oil solution. After the aqueous solution and the oil-phase encapsulation solution generate oil-in-water droplets at the intersection 5, the droplets are transported to the buffer zone 8 through the droplet flow channel 6. The oil-in-water droplets become round in the buffer zone 8, and the relative positions of different droplets are stabilized. The aqueous NAD is collected in the outlet area 7 of the buffer zone 8. + Encapsulation product of the precursor.

[0105] In some possible implementations, the inner diameter of the inlet region 1 of the aqueous solution is 800 μm to 1200 μm; the larger inner diameter is used to store and provide the aqueous solution. Exemplarily, the inner diameter of the inlet region 1 of the aqueous solution can be 800 μm, 900 μm, 1000 μm, 1200 μm, or any other typical but non-limiting point value or an interval value between any two point values.

[0106] In some possible implementations, the inner diameter of the input flow channel 2 of the aqueous solution is 10 μm to 500 μm. In this case, the inner diameter of the input flow channel 2 of the aqueous solution ensures that the aqueous solution has good flow performance in the flow channel to avoid excessive flow resistance that causes the solution to be difficult to flow; it also ensures that the aqueous solution has good surface shear force so as to form a droplet state at the intersection 5. Exemplarily, the inner diameter of the input flow channel 2 of the aqueous solution is 10 μm, 50 μm, 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, etc., typical but non-restrictive arbitrary point values ​​or interval values ​​between any two point values.

[0107] In some possible implementations, the width of the input channel 2 for the aqueous solution is 10 μm to 500 μm, and the height is 10 μm to 500 μm; and the ratio of the height to the width is 1:1, which is convenient for channel processing and avoids channel blockage.

[0108] In some possible implementations, the inner diameter of the input flow channel 2 of the aqueous solution near the intersection 5 is smaller than the inner diameter of the end near the inlet region 1 of the aqueous solution. In this case, the narrowing of the flow channel can speed up the flow rate of the solution, making it easier to form a liquid at the intersection 5 to obtain an oil-in-water structure.

[0109] In some possible implementations, in the input flow channel 2 of the aqueous solution, the ratio of the inner diameter at the end close to the intersection 5 to the inner diameter at the end close to the inlet region 1 of the aqueous solution is (1.5-2.5):1. In this case, it is ensured that the aqueous solution has good flow properties in the flow channel, and it is also conducive to forming a liquid at the intersection 5 to obtain an oil-in-water structure. Exemplarily, in the input flow channel 2 of the aqueous solution, the ratio of the inner diameter at the end close to the intersection 5 to the inner diameter at the end close to the inlet region 1 of the aqueous solution can be 1.5:1, 2:1, 2.5:1, or other typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values.

[0110] In some possible implementations, the inner diameter of the inlet region 3 for the encapsulated solution is 800 μm to 1200 μm; the larger inner diameter is used to store and provide the oil phase encapsulated solution. Exemplarily, the inner diameter of the inlet region 3 for the encapsulated solution can be 800 μm, 900 μm, 1000 μm, 1200 μm, or any other typical but non-limiting point value or an interval value between any two point values.

[0111] In some possible implementations, the inner diameter of the input flow channel 4 of the encapsulated solution is 10 μm to 500 μm. In this case, the inner diameter of the input flow channel 4 of the encapsulated solution ensures that the encapsulated solution has good flow performance in the flow channel to avoid excessive flow resistance that causes the solution to be difficult to flow; it also ensures that the encapsulated solution has good surface shear force so that a droplet state is formed at the intersection 5. Exemplarily, the inner diameter of the input flow channel 4 of the encapsulated solution can be 10 μm, 50 μm, 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, etc., typical but non-restrictive arbitrary point values ​​or interval values ​​between any two point values.

[0112] In some possible implementations, the width of the input channel 4 for the encapsulation solution is 10 μm to 500 μm, and the height is 10 μm to 500 μm; and the ratio of the height to the width is 1:1, which facilitates channel processing and avoids channel blockage.

[0113] In some possible implementations, in the input flow channel 4 of the encapsulated solution, the inner diameter of the end near the intersection 5 is smaller than the inner diameter of the end near the inlet area 3 of the encapsulated solution. In this case, the narrowing of the flow channel can speed up the flow rate of the solution, making it easier to form a liquid at the intersection 5 to obtain an oil-in-water structure.

[0114] In some possible implementations, in the input flow channel 4 of the encapsulated solution, the ratio of the inner diameter at the end close to the intersection 5 to the inner diameter at the end close to the inlet area 3 of the encapsulated solution is (1.5-2.5):1. In this case, it is ensured that the encapsulated solution has good flow properties in the flow channel, and it is also conducive to forming a liquid at the intersection 5 to obtain an oil-in-water structure. Exemplarily, in the input flow channel 4 of the encapsulated solution, the ratio of the inner diameter at the end close to the intersection 5 to the inner diameter at the end close to the inlet area 3 of the encapsulated solution can be 1.5:1, 2:1, 2.5:1, or other typical but non-limiting arbitrary point values ​​or interval values ​​between any two point values.

[0115] In some possible implementations, the ratio of the inner diameter of the input flow channel 2 of the aqueous solution to the inner diameter of the input flow channel 4 of the encapsulation solution is 1:1. In this case, it is more conducive to controlling the generation of an encapsulated product with an oil-in-water structure by regulating the flow rates of the aqueous solution and the oil-phase encapsulation solution, thereby preventing the aqueous solution and the oil-phase encapsulation solution from forming a laminar flow at the intersection 5 and failing to form droplets.

[0116] In some possible implementations, the ratio of the length of the flow channel of the buffer zone 8 to the length of the droplet flow channel 6 is (2-10):1. In this case, the flow channel of the buffer zone 8 has a suitable length, which is conducive to rounding the micro-droplets and stabilizing the relative positions of different micro-droplets, avoiding the influence of the stability of the droplets due to the excessive length of the flow channel and the high flow resistance. Exemplarily, the ratio of the length of the flow channel of the buffer zone 8 to the length of the droplet flow channel 6 can be 2:1, 2.5:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, etc.

[0117] In some possible implementations, the inner diameter of the droplet flow channel 6 is 10 μm to 500 μm, and can be, for example, 10 μm, 50 μm, 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, or any other typical but non-limiting point value or an interval value between any two point values. In this case, the inner diameter of the droplet flow channel 6 is conducive to maintaining the fluidity of the water-in-oil droplet while ensuring the stability of the water-in-oil droplet structure.

[0118] In some possible implementations, the inner diameter of the flow channel of the buffer zone 8 is 10 μm to 500 μm, and can be, for example, any typical but non-restrictive point value or an interval value between any two point values ​​such as 10 μm, 50 μm, 100 μm, 200 μm, 300 μm, 400 μm, and 500 μm. In this case, the inner diameter of the flow channel of the buffer zone 8 can round the droplets and stabilize the relative positions of different micro-droplets, thereby avoiding the influence of the stability of the droplets on the flow channel being too long and the flow resistance being too high.

[0119] In a second aspect, the present invention provides an aqueous NAD + Encapsulation product of precursor, aqueous NAD + The encapsulated product of the precursor is an oil-in-water structure, including NAD + The precursor has a water phase core and an oil phase encapsulation layer, wherein the oil phase encapsulation layer comprises an oil phase material and a surfactant.

[0120] The aqueous phase NAD of the present application example + The encapsulated product of the precursor is an oil-in-water structure, including NAD + The precursor has a water phase core and an oil phase encapsulation layer, and the oil phase encapsulation layer contains oil phase materials and surfactants. The coating of the oil phase encapsulation layer significantly increases the NAD + Stability of precursors, effective protection of NAD + The precursor is not affected by factors such as oxygen, ultraviolet light, temperature and pH in the environment, avoiding the presence of aqueous NAD + Precursor inactivation significantly increases aqueous NAD + Stability of precursors during storage, transportation and application.

[0121] The aqueous phase NAD of the present application example + The encapsulated product of the precursor can be prepared by the aqueous NAD + The precursor is prepared by the encapsulation method.

[0122] In some possible implementations, the aqueous phase NAD + The average particle size of the encapsulated product of the precursor is 50 μm to 100 μm, and can be, for example, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, or any other typical but non-limiting point value or an interval value between any two point values. Figure 6 In this case, the aqueous NAD + The encapsulation product of the precursor has a small particle size, which is beneficial to the core NAD + The precursor is released and has a larger contact specific surface area, which is more conducive to its application and activity in the fields of health care products, skin care products, etc.

[0123] In some possible implementations, the aqueous phase NAD + In the encapsulated product of the precursor, the mass ratio of the water phase core to the oil phase encapsulation layer is (20-35): (50-65). In this case, the mass ratio of the water phase core to the oil phase encapsulation layer is conducive to maintaining the stability of the oil-in-water structure of the encapsulated product. The oil phase encapsulation layer can effectively isolate the factors such as oxygen, ultraviolet light, temperature and pH in the environment from affecting the water phase NAD + The interference of precursor stability improves the environmental stability of the encapsulated product. + In the encapsulated product of the precursor, the mass ratio of the water phase core to the oil phase encapsulation layer can be any typical but non-limiting point value such as 20:50, 25:55, 30:60, 35:65, or an interval value between any two points.

[0124] In some possible implementations, the aqueous core also contains an antioxidant and a buffer solution; by adding the antioxidant and the buffer solution, oxidation reactions can be prevented and a suitable pH environment can be maintained, so that NAD + The precursor is not affected by oxidation and pH changes, thus protecting NAD in aqueous solutions + Activity of precursors.

[0125] In some possible implementations, the outer surface of the oil phase encapsulation layer further includes a polymer carrier layer, and the thickness of the polymer carrier layer is 50nm to 200nm, and can be, for example, 50nm, 100nm, 150nm, 200nm, or any other typical but non-limiting point value or an interval value between any two point values. In this case, the polymer carrier layer can further maintain the aqueous phase NAD + The structural stability and environmental stability of the encapsulated product of the precursor are beneficial to the storage, transportation, application and other processes.

[0126] In some possible implementations, the aqueous phase NAD + The encapsulated product of the precursor is stored in a sealed, light-proof environment at a temperature of -20°C to 25°C. Exemplarily, it can be any typical but non-limiting point value or an interval value between any two points such as -20°C, -10°C, -5°C, 0°C, 5°C, 10°C, 15°C, 20°C, 25°C. This low-temperature sealed, light-proof environment is more conducive to maintaining the aqueous phase NAD + Stability of the encapsulated product of the precursor.

[0127] In a third aspect, the present invention provides an aqueous NAD + The encapsulation product of the precursor is used to prepare the aqueous NAD + The encapsulated product of the precursor or the aqueous NAD + The encapsulated product of the precursor is applied to at least one of the fields of skin care products, health care products and medicines.

[0128] The aqueous phase NAD of the present application example + The encapsulation product of the precursor contains NAD stably + Precursors have the functions of maintaining cell function, delaying biological aging, enhancing immunity, improving cognitive ability, protecting the liver and vision, and removing free radicals that may cause cell aging. They can not only be used to develop skin care products with anti-aging effects, but also can be used in health products and medicines with the effects of reverse aging, enhancing vitality, enhancing immunity, protecting the liver and vision, improving athletic ability and rapid recovery.

[0129] In order to make the above implementation details and operations of this application clearly understood by those skilled in the art, as well as the aqueous phase NAD of this application example + The encapsulated product of the precursor and its encapsulation method and application have significantly improved performance. The above technical solution is illustrated by multiple embodiments below.

[0130] Example 1

[0131] A water-phase NAD(P)H encapsulation product, wherein the encapsulation method comprises the following steps:

[0132] 1. Prepare the following by photolithography Figure 2 The microfluidic chip shown: photoresist is coated on the silicon wafer, and the desired microfluidic channel pattern is created by photolithography technology; polydimethylsiloxane (PDMS) and curing agent are mixed in a ratio of 10:1, stirred evenly and poured on the silicon wafer; cured at 80°C for 30 minutes; after curing, the PDMS is peeled off the silicon wafer and plasma bonded to a glass sheet or another PDMS layer to form a complete microfluidic chip.

[0133] 2. Prepare an aqueous solution of NAD(P)H: dissolve 10 g / L NAD(P)H and 5% w / v vitamin C in 0.1 M Tris buffer (pH 7.4).

[0134] 3. Prepare oil phase encapsulation solution: Dissolve 4% w / v surfactant T154 in soybean oil.

[0135] 4. Use attached Figure 2 In the microfluidic chip shown, the aqueous solution of NAD(P)H and the oil-phase encapsulation solution are respectively injected into the aqueous solution inlet area 1 (radius of 1000 μm) and the oil-phase encapsulation solution inlet area 3 (radius of 1000 μm) of the microfluidic chip by a syringe pump. The aqueous solution is transported to the intersection 5 at a flow rate of 20 μL / h through the aqueous solution input channel 2 (the inner diameter of the inlet area 1 near the aqueous solution is 100 μm, and the inner diameter of the inlet area near the intersection 5 is 50 μm), and the oil-phase encapsulation solution is transported to the intersection 5 at a flow rate of 50 μL / h through the encapsulation solution input channel 4 (the inner diameter of the inlet area 3 near the encapsulation solution is 100 μm, and the inner diameter of the inlet area 5 near the intersection 5 is 50 μm). Oil-in-water droplets are generated at the intersection 5T-shaped structure; the generated oil-in-water droplets are transported to the buffer zone 8 through the droplet flow channel 6 (inner diameter 50 μm, length 9500 μm), and the oil-in-water droplets become round in the flow channel (inner diameter 50 μm, length 199100 μm) of the buffer zone 8, and the relative positions of different droplets are stabilized, and the encapsulated product of aqueous phase NAD(P)H is collected in the outlet area 7 of the buffer zone 8, and the particle size is about 50 μm.

[0136] Example 2

[0137] A water-phase NAD(P)H encapsulation product, wherein the encapsulation method comprises the following steps:

[0138] 1. Use 3D printing technology to prepare the attached Figure 3 The microfluidic chip shown: a microfluidic channel pattern is prepared by 3D printing technology; polydimethylsiloxane (PDMS) and a curing agent are mixed in a ratio of 10:1, stirred evenly and poured onto a silicon wafer; cured at 80°C for 30 minutes; after curing, the PDMS is peeled off the silicon wafer and plasma bonded to a glass sheet or another PDMS layer to form a complete microfluidic chip.

[0139] 2. Prepare an aqueous solution of NAD(P)H: Dissolve 10 g / L NAD(P)H and 3% w / v glutathione in 0.1 M phosphate buffered saline (PBS) (pH 7.4).

[0140] 3. Prepare oil phase encapsulation solution: Dissolve gelatin in warm water, then slowly add it to light mineral oil, stir evenly, then add surfactant Span 80, stir evenly. Gelatin can form a flexible membrane, which helps to isolate air, maintain droplet shape and control release. The content is 1% (w / v), and Span 80 acts as a surfactant, with a content of 2% (w / v).

[0141] 4. Use attached Figure 3 In the microfluidic chip shown, the aqueous solution of NAD(P)H and the oil-phase encapsulation solution are respectively injected into the aqueous solution inlet area 1 (radius of 1000 μm) and the oil-phase encapsulation solution inlet area 3 (radius of 1000 μm) of the microfluidic chip by a syringe pump. The aqueous solution is transported to the intersection 5 at a flow rate of 35 μL / h through the aqueous solution input channel 2 (the inner diameter of the inlet area 1 near the aqueous solution is 100 μm, and the inner diameter of the inlet area 5 near the intersection 5 is 50 μm), and the oil-phase encapsulation solution is transported to the intersection 5 at a flow rate of 65 μL / h through the encapsulation solution input channel 4 (the inner diameter of the inlet area 3 near the encapsulation solution is 100 μm, and the inner diameter of the inlet area 5 near the intersection 5 is 50 μm). Oil-in-water droplets are generated at the intersection 5 T-shaped structure; the generated oil-in-water droplets become round in the droplet flow channel 6 (with an inner diameter of 50 μm and a length of 9500 μm), and the relative positions of different droplets are stabilized. The encapsulated products of aqueous phase NAD(P)H are collected in the outlet area 7, and the particle size is about 50 μm. Figure 6 shown.

[0142] Example 3

[0143] A water-phase NAD(P)H encapsulation product, wherein the encapsulation method comprises the following steps:

[0144] 1. Prepare the following by photolithography Figure 4 The microfluidic chip shown: photoresist is coated on the silicon wafer, and the desired microfluidic channel pattern is created by photolithography technology; polydimethylsiloxane (PDMS) and curing agent are mixed in a ratio of 10:1, stirred evenly and poured on the silicon wafer; cured at 80°C for 30 minutes; after curing, the PDMS is peeled off the silicon wafer and plasma bonded to a glass sheet or another PDMS layer to form a complete microfluidic chip.

[0145] 2. Prepare the aqueous solution of NAD(P)H: dissolve 1 g / L NAD(P)H in Tris buffer at pH 7.4, add 0.5% glutathione, and mix well.

[0146] 3. Prepare the oil phase encapsulation solution: dissolve 5% tetraethyl orthosilicate (TEOS) in argan oil, add 2% surfactant Span 80 and 0.01M hydrochloric acid, and stir evenly.

[0147] 4. Use attached Figure 4 In the microfluidic chip shown, the aqueous solution of NAD(P)H and the oil-phase encapsulation solution are respectively injected into the aqueous solution inlet area 1 (radius of 1000 μm) and the oil-phase encapsulation solution inlet area 3 (radius of 1000 μm) of the microfluidic chip by a syringe pump. The aqueous solution is transported to the intersection 5 at a flow rate of 35 μL / h through the aqueous solution input channel 2 (the inner diameter of the inlet area 1 near the aqueous solution is 100 μm, and the inner diameter of the inlet area 5 near the intersection 5 is 50 μm), and the oil-phase encapsulation solution is transported to the intersection 5 at a flow rate of 65 μL / h through the encapsulation solution input channel 4 (the inner diameter of the inlet area 3 near the encapsulation solution is 100 μm, and the inner diameter of the inlet area 5 near the intersection 5 is 50 μm). Oil-in-water droplets are generated at the intersection 5T-shaped structure; the generated oil-in-water droplets become rounded in the droplet flow channel 6 (inner diameter 50 μm, length 9500 μm), and the relative positions of different droplets are stabilized, and the encapsulated products of aqueous phase NAD(P)H are collected in the outlet area 7, with a particle size of about 50 μm.

[0148] 5. After the encapsulated droplets of aqueous NAD(P)H are exposed to air, ethyl orthosilicate is hydrolyzed and condensed under the catalysis of hydrochloric acid to form a silica shell, thereby obtaining an encapsulated product of aqueous NAD(P)H.

[0149] In order to verify the progress of the embodiments of the present application, the following performance tests were performed on the above embodiments:

[0150] 1. Droplet uniformity test: The droplet shape, breakage and dispersion of the encapsulated product of aqueous NAD(P)H collected in outlet area 7 of Example 2 were observed using an optical microscope and a high-speed camera.

[0151] The test results are as attached Figure 6 As shown, the droplet diameter is about 50 μm and the droplet size is 1962 ± 178 μm 2 , the droplet uniformity is good; where the droplet size refers to the area of ​​the droplet in the figure, which is measured using ImageJ software.

[0152] 2. Stability test: After the encapsulated product droplets of NAD(P)H prepared in the above embodiments were stored at 4°C and room temperature for 1 day, 3 days, 1 week, 2 weeks, 3 weeks and 1 month, the concentration of NAD(P)H was measured regularly using an ultraviolet spectrophotometer. The test results are shown in Table 1 below:

[0153] Table 1

[0154]

[0155]

[0156] From the above test results, it can be seen that the encapsulated product of the embodiment of the present application is an oil-in-water structure, and the aqueous phase NAD + The precursor NAD(P)H is encapsulated in the oil phase shell layer, isolating the interference of environmental factors such as oxygen, ultraviolet light, temperature and pH on the stability of aqueous phase NAD(P)H, significantly improving the stability of aqueous phase NAD + Stability of the precursor at low temperature (4 °C) and room temperature.

[0157] 3. Biological activity test: The NAD(P)H colorimetric detection kit was used to measure the biological activity of the encapsulated product of NAD(P)H in the embodiment when droplets were generated and stored for 1 day, 3 days, 1 week, and 2 weeks. The test results are shown in Table 2 below:

[0158] Table 2

[0159] Biological activity test Example 1 Example 2 Example 3 When generated 100％ 100％ 100％ 1 day 100％ 100％ 100％ 3 days 99.3％ 99.7％ 99.8％ 1 week 98.6％ 99.2％ 99％ 2 weeks 97.9％ 98％ 97.7％

[0160] From the above test results, it can be seen that the aqueous phase NAD encapsulated product of the present embodiment + The precursor NAD(P)H is encapsulated into the oil phase shell layer, isolating the interference of environmental factors such as oxygen, ultraviolet light, temperature and pH on the stability of aqueous phase NAD(P)H, and significantly improving the aqueous phase NAD in the encapsulated product. + The storage biological activity of the precursor NAD(P)H facilitates its flexible application in areas such as skin care and health products.

[0161] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. Aqueous NAD + A method for encapsulating a precursor, characterized in that The following steps are involved: Preparation containing NAD + an aqueous solution of a precursor; preparing an encapsulation solution comprising an oil phase material and a surfactant; Prepare a microfluidic chip, use the microfluidic chip to make the encapsulation solution and the aqueous solution into an oil-in-water core-shell structure through microfluidic technology, and obtain aqueous NAD + Encapsulation product of the precursor.

2. The aqueous NAD as claimed in claim 1 + A method for encapsulating a precursor, characterized in that The NAD + The precursor includes at least one of NAD(P)H, NMN, and NR; and / or, the oil phase material comprises at least one of mineral oil, vegetable oil, silicone oil, fluorinated oil, and synthetic oil; and / or, the surfactant comprises at least one of Span 80, Tween 20, sodium lauryl sulfate, sodium lauryl ether sulfate, cetyltrimethylammonium chloride, polyoxyethylene lauryl alcohol, polyoxyethylene phosphate, olive oil soap, and glucoside; And / or, the aqueous phase solution further comprises an antioxidant and a buffer solution; And / or, the encapsulation solution further comprises a polymer carrier or a polymer carrier and a curing agent.

3. The aqueous NAD as claimed in claim 2 + A method for encapsulating a precursor, characterized in that The NAD + The precursor is selected from NAD(P)H; And / or, in the aqueous phase solution, the concentration of the buffer solution is 0.01 mol / L to 1 mol / L, the pH value is 7.0 to 7.5, and the NAD + The concentration of the precursor is 0.1mmol / L to 10mmol / L, and the mass percentage of the antioxidant is 1% to 20%; And / or, the antioxidant includes at least one of vitamin C, vitamin E, anthocyanin, chlorophyll, carotene, and glucose; And / or, the buffer solution includes at least one of a phosphate buffered saline solution and a tris buffer solution; And / or, in the encapsulation solution, the mass percentage of the oil phase material is 80% to 95%, and the mass percentage of the surfactant is 0.02% to 5%; and / or, in the encapsulation solution, the addition amount of the polymer carrier is 0.05% to 10% of the mass of the oil phase material; and / or, in the encapsulation solution, the amount of the curing agent added is 0.05% to 10% of the mass of the oil phase material; And / or, the polymer carrier includes at least one of alginate, polysaccharide, polylactic acid-glycolic acid copolymer, and protein; And / or, the curing agent includes at least one of epoxy resin, polyamine curing agent, acrylate monomer, 2-hydroxy-2-methylacetone, protein, polysaccharide, glutaraldehyde, sodium alginate, and calcium ion.

4. The aqueous NAD according to any one of claims 1 to 3 + A method for encapsulating a precursor, characterized in that The method for preparing the microfluidic chip includes: at least one of photolithography technology and 3D printing technology; And / or, the microfluidic chip includes an inlet area of ​​an aqueous solution, an inlet area of ​​an encapsulation solution, an intersection and an outlet area, wherein the inlet area of ​​the aqueous solution is connected to the intersection through an input flow channel of the aqueous solution, the inlet area of ​​the encapsulation solution is connected to the intersection through an input flow channel of the encapsulation solution, and the intersection is connected to the outlet area through a droplet flow channel.

5. The aqueous NAD as claimed in claim 4 + A method for encapsulating a precursor, characterized in that The steps of the microfluidic technology include: transporting the aqueous phase solution and the encapsulation solution from the inlet area to the intersection through the input flow channel, generating oil-in-water droplets at the intersection through the microfluidic technology, and the oil-in-water droplets are collected in the outlet area through the droplet flow channel to obtain the aqueous phase NAD + Encapsulation product of the precursor.

6. The aqueous NAD as claimed in claim 5 + A method for encapsulating a precursor, characterized in that The method of generating water-in-oil droplets by microfluidics technology includes one of T-type structure, flow focusing, cross flow, coaxial flow, and step emulsification; and / or, the ratio of the flow rates of the aqueous solution and the encapsulation solution reaching the intersection is (4-7):(10-13); and / or, the flow rate of the aqueous solution reaching the intersection is 20 μL / h to 35 μL / h; And / or, the flow rate of the encapsulation solution reaching the intersection is 50 μL / h to 65 μL / h.

7. The aqueous NAD as claimed in claim 5 + A method for encapsulating a precursor, characterized in that In the microfluidic chip, a buffer zone is further provided between the droplet flow channel and the outlet area; and / or, the inner diameter of the inlet region of the aqueous solution is 800 μm to 1200 μm; and / or, the inner diameter of the inlet region of the encapsulated solution is 800 μm to 1200 μm; And / or, the inner diameter of the input flow channel of the aqueous solution is 10 μm to 500 μm; and / or, the inner diameter of the input channel of the encapsulation solution is 10 μm to 500 μm; And / or, the inner diameter of the droplet flow channel is 10 μm to 500 μm; And / or, the inner diameter of the flow channel of the buffer zone is 10 μm to 500 μm.

8. The aqueous NAD as claimed in claim 7 + A method for encapsulating a precursor, characterized in that The ratio of the length of the flow channel of the buffer zone to the length of the droplet flow channel is (2-10):1; And / or, in the input flow channel of the aqueous solution, the inner diameter of the end close to the intersection is smaller than the inner diameter of the end close to the inlet region of the aqueous solution; And / or, in the input flow channel of the encapsulated solution, the inner diameter of the end close to the intersection is smaller than the inner diameter of the end close to the inlet area of ​​the encapsulated solution.

9. The aqueous NAD as claimed in claim 8 + A method for encapsulating a precursor, characterized in that In the input flow channel of the aqueous solution, the ratio of the inner diameter of the end close to the intersection to the inner diameter of the end close to the inlet area of ​​the aqueous solution is (1.5-2.5):1; And / or, in the input flow channel of the encapsulation solution, the ratio of the inner diameter of the end close to the intersection to the inner diameter of the end close to the inlet area of ​​the encapsulation solution is (1.5-2.5):1; And / or, the ratio of the inner diameter of the input flow channel of the aqueous solution to the inner diameter of the input flow channel of the encapsulation solution is 1:

1.

10. Aqueous NAD + The encapsulated product of the precursor is characterized in that The aqueous NAD + The encapsulated product of the precursor is an oil-in-water structure, including NAD + The precursor has a water phase inner core and an oil phase encapsulation layer, wherein the oil phase encapsulation layer comprises an oil phase material and a surfactant.

11. The aqueous NAD according to claim 10 + The encapsulated product of the precursor is characterized in that The aqueous NAD + The average particle size of the encapsulated product of the precursor is 50 μm to 100 μm; And / or, the aqueous phase NAD + In the encapsulated product of the precursor, the mass ratio of the water phase core to the oil phase encapsulation layer is (20-35): (50-65); And / or, the aqueous core further comprises an antioxidant and a buffer solution; And / or, the outer surface of the oil phase encapsulation layer further comprises a polymer carrier layer, and the thickness of the polymer carrier layer is 50nm to 200nm; And / or, the aqueous phase NAD + The encapsulated product of the precursor is stored in a sealed, light-proof environment at a temperature of -20°C to 25°C.

12. Aqueous NAD + Application of the encapsulated product of the precursor, characterized in that The aqueous NAD prepared by the method according to any one of claims 1 to 9 + The encapsulated product of the precursor or the aqueous phase NAD as claimed in any one of claims 10 to 11 + The encapsulated product of the precursor is applied to at least one of the fields of skin care products, health care products and medicines.

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