Preparation method and application of NMN oil suspension

Abstract

The present application relates to a kind of preparation method and application of NMN oil suspension, belong to the field of cosmetics.The NMN oil suspension of the present application is prepared by β-nicotinamide mononucleotide, water-soluble functional ingredient powder, oil phase emollient.The β-nicotinamide mononucleotide of the present application is encapsulated with water-soluble functional ingredient by oil phase, can significantly improve the stability of β-nicotinamide mononucleotide, prolong the shelf life of product containing β-nicotinamide mononucleotide.The β-nicotinamide mononucleotide of the present application is compounded with water-soluble functional ingredient, can also improve the anti-aging efficacy of composition, realize synergistic effect, reduce use cost.The NMN oil suspension prepared in the present application does not use emulsifier, is more mild, can be applied to prepare stable anti-aging cosmetic.

Description

Technical Field

[0001] This invention relates to a method for preparing and applying an NMN oil suspension, belonging to the field of cosmetic technology. Background Technology

[0002] NMN, also known as β-nicotinamide mononucleotide, is a precursor to NAD+, a cofactor of longevity proteins in the human body. NAD+ is not only an important raw material for cell repair but also a key communication factor between the cell nucleus and mitochondria responsible for energy synthesis. Its content is closely related to the activity of genes in the human body that have the function of prolonging life and inhibiting aging. Since NAD+ molecules are difficult to enter cells directly, supplementing NAD+ means supplementing NAD+ precursors to be absorbed into the bloodstream so that it can be utilized and converted within cells.

[0003] Chinese patent document CN115531216A (application number 202211229539.5) discloses a soluble microneedle patch, in which the active ingredients are selected from NMN and adenosine. This invention opens the stratum corneum channels through soluble microneedle technology, replenishing beneficial active skin components and promoting absorption and efficacy. NMN can increase cell vitality and enhance cell absorption of active ingredients; adenosine has excellent moisturizing effects and is an important intermediate in the synthesis of cellular energy components. The combined use of NMN and adenosine in soluble microneedles within a hyaluronic acid matrix can increase mitochondrial energy levels and enhance cell water-locking ability, thereby ensuring long-lasting skin hydration and playing an anti-aging role. However, in this patent document, the active ingredients NMN and adenosine are mainly dissolved in the aqueous phase. In practical applications, we found that in stability studies of NMN aqueous solutions at room temperature, light exposure, and high temperature (45°C), β-nicotinamide mononucleotide (NMN) continuously decomposes and becomes inactive over time, which greatly limits its application in formulations.

[0004] Therefore, how to maintain the stability of β-nicotinamide mononucleotide (NMN) while also ensuring its anti-aging effects is a problem we need to solve. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a method for preparing and applying an NMN oil suspension. The NMN oil suspension is prepared from β-nicotinamide mononucleotide (NMN), water-soluble active ingredient powder, and an oil-phase emollient. This NMN oil suspension, by encapsulating β-nicotinamide mononucleotide and the water-soluble active ingredient with an oil-phase emollient, can significantly improve the stability of β-nicotinamide mononucleotide (NMN) and extend the shelf life of products containing β-nicotinamide mononucleotide.

[0006] To achieve the above-mentioned technical objectives, the present invention provides the following technical solution:

[0007] In a first aspect, the present invention provides an NMN oil suspension composition, wherein the components of the NMN oil suspension composition are, in parts by weight: 0.5 to 6 parts of β-nicotinamide mononucleotide (NMN), 0.2 to 3 parts of water-soluble active ingredient, and an oil phase emollient to make up to 100 parts.

[0008] Preferably, the NMN oil suspension composition comprises, in parts by weight: 5 parts of β-nicotinamide mononucleotide (NMN), 1.8 parts of water-soluble active ingredient, and an oil phase emollient to make up to 100 parts.

[0009] Preferably, the β-nicotinamide mononucleotide (NMN) is in powder form with a particle size between 400 and 600 nm and a regular spherical microstructure.

[0010] Preferably, the water-soluble active ingredient is in powder form, with a particle size between 800 and 900 nm and a regular spherical microstructure.

[0011] Preferably, the water-soluble active ingredient is at least one of adenosine, ferulic acid, vitamin C and its derivatives.

[0012] Preferably, the oil-phase emollient is one or more of the following: isopropyl palmitate, isopropyl myristate, isooctyl palmitate, dioctyl carbonate, caprylic / capric triglyceride, squalane, meadowfoam seed oil, sunflower seed oil, vitamin E acetate, hydrogenated polyisobutylene, isononyl isononanoate, C12-15 alcohol benzoate, marula oil, emu oil, sweet almond oil, C9-12 alkyl, cocoyl alcohol-caprylic / capric ester, polydimethylsiloxane, plant squalane, isooctyl palmitate, and isopropyl myristate, but not limited to commonly available oil components.

[0013] A second aspect of the present invention provides a method for preparing an NMN oil suspension, comprising the following steps:

[0014] S1. Under normal temperature conditions, β-nicotinamide mononucleotide and water-soluble active ingredient are mechanically ball-milled to obtain β-nicotinamide mononucleotide powder and water-soluble active ingredient powder that meet the particle size requirements.

[0015] S2. Under normal temperature conditions, β-nicotinamide mononucleotide powder is added to 1 / 2 to 3 / 4 of the oil phase emollient and dispersed by high-speed shearing, and then homogenized under high pressure to obtain oil 1;

[0016] S3. Under normal temperature conditions, the water-soluble active ingredient powder is added to the remaining oil phase emollient and dispersed by high-speed shearing, and then homogenized under high pressure to obtain oil 2.

[0017] S4. Mix oil 1 and oil 2, stir and disperse evenly at room temperature, and then homogenize under high pressure to obtain NMN oil suspension.

[0018] In the above preparation method, steps S2 and S3 do not have a specific order.

[0019] In the above preparation method, the high-speed shear dispersion is achieved using a high-shear homogenizing emulsifier, and the high-pressure homogenization is achieved using a high-pressure microfluidic homogenizer.

[0020] Preferably, in step S2, the high-pressure homogenization process involves a homogenization rate of 6000–8000 rpm, a homogenization time of 8–12 min, and a homogenization pressure of 1.4–1.6 MPa; the process is repeated 3–6 times.

[0021] More preferably, in step S2, the high-pressure homogenization is performed at a homogenization rate of 7500 rpm, a homogenization time of 10 min, and a homogenization pressure of 1.5 MPa; the cycle is repeated 4 times.

[0022] Preferably, in step S3, the high-pressure homogenization process involves a homogenization rate of 4000–6000 rpm, a homogenization time of 6–8 min, and a homogenization pressure of 1.0–1.2 MPa; the process is repeated 2–4 times.

[0023] More preferably, in step S3, the high-pressure homogenization is performed at a homogenization rate of 5000 rpm, a homogenization time of 7 min, and a homogenization pressure of 1.1 MPa; the cycle is repeated 3 times.

[0024] Preferably, in step S4, the high-pressure homogenization is performed at a homogenization rate of 4000-6000 rpm, a homogenization time of 4-6 min, and a homogenization pressure of 1.2-1.4 MPa; the cycle is repeated 2-4 times.

[0025] More preferably, in step S4, the high-pressure homogenization is performed at a homogenization rate of 5500 rpm, a homogenization time of 5 min, and a homogenization pressure of 1.3 MPa; the cycle is repeated twice.

[0026] In a third aspect, the present invention also provides a cosmetic product with anti-aging effects, the cosmetic product comprising the above-mentioned NMN oil suspension.

[0027] In this invention, the term "cosmetics" should be interpreted broadly, including but not limited to face creams, lotions, two-component toners, two-component masks, and two-component serums.

[0028] Beneficial effects:

[0029] 1. This invention combines β-nicotinamide mononucleotide with a water-soluble active ingredient (adenosine) and encapsulates it in an oil phase, which can significantly improve the stability of β-nicotinamide mononucleotide (NMN) and extend the shelf life of products containing β-nicotinamide mononucleotide.

[0030] 2. This invention combines β-nicotinamide mononucleotide (NMN) with water-soluble active ingredients, which can enhance the anti-aging effect of the composition, achieve synergistic effects, and reduce the cost of use.

[0031] 3. The NMN oil suspension prepared by this invention does not use emulsifiers, making it gentler and suitable for use in the preparation of stable anti-aging cosmetics. Detailed Implementation

[0032] The technical solution of the present invention will be further described below with reference to specific embodiments. Unless otherwise specified, the raw materials involved in the embodiments are all commercially available products.

[0033] Examples 1-4:

[0034] The NMN oil suspension composition includes β-nicotinamide mononucleotide (NMN), a water-soluble active ingredient, and an oil-phase emollient. Both β-nicotinamide mononucleotide (NMN) and the water-soluble active ingredient are in powder form. The particle size of the β-nicotinamide mononucleotide (NMN) powder is between 400 and 600 nm, and its microstructure is a regular spherical shape. The particle size of the water-soluble active ingredient powder is between 800 and 900 nm, and its microstructure is also a regular spherical shape.

[0035] The composition of the NMN oil suspension compositions in Examples 1-4 and the preparation process parameters of the NMN oil suspensions are shown in Table 1, all as mass percentages. The water-soluble active ingredient in the NMN oil suspension compositions is adenosine, and the oil-phase emollient is caprylic / capric triglyceride.

[0036] Table 1. Composition of NMN oil suspension compositions in Examples 1-4 and preparation process parameters of NMN oil suspensions

[0037]

[0038] The preparation methods of NMN oil suspensions in Examples 1-4 include the following steps:

[0039] S1. Under normal temperature conditions, β-nicotinamide mononucleotide and adenosine are mechanically ball-milled to obtain β-nicotinamide mononucleotide powder and adenosine powder that meet the particle size requirements.

[0040] S2. Under normal temperature conditions, β-nicotinamide mononucleotide powder was added to 1 / 2 caprylic / capric triglyceride, and high-speed shear dispersion was performed using a high-shear homogenizer emulsifier. Then, high-pressure homogenization was performed using a high-pressure microfluidic homogenizer according to the process parameters in Table 1 to obtain oil 1.

[0041] S3. Under normal temperature conditions, adenosine powder is added to the remaining 1 / 2 of caprylic / capric triglyceride, and high-speed shear dispersion is performed using a high-shear homogenizer emulsifier. Then, high-pressure homogenization is performed using a high-pressure micro-jet homogenizer according to the process parameters in Table 1 to obtain oil 2.

[0042] S4. Mix oil 1 and oil 2, stir and disperse evenly under normal temperature conditions, and then use a high-pressure micro-jet homogenizer to homogenize under high pressure according to the process parameters in Table 1 to obtain NMN oil suspension.

[0043] Examples 5-14:

[0044] The NMN oil suspension compositions of Examples 5-14 were formed by replacing the oil phase emollient caprylic / capric triglyceride in Example 1 with squalane, meadowfoam seed oil, sunflower seed oil, hydrogenated polyisobutylene, isononyl isononanoate, C12-15 alcohol benzoate, C9-12 alkyl, coconut oil alcohol-caprylic / capric triglyceride, polydimethylsiloxane, and plant squalane, respectively. All other components were the same as in Example 1.

[0045] The preparation methods of NMN oil suspensions in Examples 5-14 are the same as those in Example 1.

[0046] Comparative Examples 1-9:

[0047] The compositions of the oil suspension compositions in Comparative Examples 1-9 are shown in Table 2, all in mass percentage.

[0048] Table 2. Composition of the oil suspension compositions in Comparative Examples 1-9

[0049] β-Nicotinamide mononucleotide (NMN) adenosine Caprylic / Capric Triglyceride Comparative Example 1 5.00％ / to 100 Comparative Example 2 / 1.80％ to 100 Comparative Example 3 10.00％ / to 100 Comparative Example 4 / 5.00％ to 100 Comparative Example 5 5.00％ Replace with 1.80% hydrophobic silica. to 100 Comparative Example 6 5% (particle size greater than 600nm) 1.80% (particle size less than 800nm) to 100 Comparative Example 7 5% (particle size less than 400nm) 1.80% (particle size greater than 900 nm) to 100 Comparative Example 8 5% (particle size less than 400nm) 1.80% (particle size less than 800nm) to 100 Comparative Example 9 5% (particle size greater than 600nm) 1.80% (particle size greater than 900 nm) to 100

[0050] The preparation methods of the oil suspensions in Comparative Examples 1 to 9 are the same as those in Example 1.

[0051] Experimental Example 1: High and Low Temperature Stability Test of Oil Suspension

[0052] Test samples: oil suspension samples from Examples 1-14 and Comparative Examples 1-9.

[0053] Stability study: The oil suspension samples to be tested were placed at room temperature (25℃), under light (light intensity 4500Lx±500Lx, 24 hours of continuous light), 4℃, 45℃, -18℃, and under cycling (-18℃, 4℃, 45℃, one cycle of temperature change every 24 hours) for 1 month. The stratification of the samples was observed, and the results are shown in Table 3.

[0054] Table 3. Results of stability studies on oil suspension samples from Examples 1-14 and Comparative Examples 1-9

[0055]

[0056] Analysis of the stability test results of Examples 1 to 14 shows that the NMN oil suspension obtained by compounding β-nicotinamide mononucleotide powder with water-soluble active ingredient powder (adenosine powder) and sealing it in an oil has good stability. After being placed for one month under the conditions of room temperature, light, 4℃, 45℃, -18℃, and cyclic (-18℃, 4℃, 45℃, with a cycle of temperature change every 24 hours), it still maintains a normal appearance and no stratification phenomenon occurs.

[0057] Analysis of the stability test results of Example 1 and Comparative Examples 1-4 shows that when β-nicotinamide mononucleotide powder or water-soluble active ingredient powder (adenosine powder) is sealed alone, the resulting oil suspension exhibits stratification and poor stability. After one month of storage under conditions of room temperature, light exposure, 4℃, 45℃, -18℃, and cyclic temperature changes (-18℃, 4℃, 45℃, with a cycle every 24 hours), the stratification phenomenon becomes more pronounced. These results indicate that the addition of water-soluble active ingredient powder (adenosine powder) is beneficial to the stability of the NMN oil suspension system, and that the combined use of β-nicotinamide mononucleotide powder and water-soluble active ingredient powder (adenosine powder) has a certain synergistic stabilizing ability.

[0058] Analysis of the stability test results of Example 1 and Comparative Example 5 shows that although the oil suspension prepared by replacing adenosine with hydrophobic silica appears normal, the oil suspension system is unstable. After being placed for one month under the conditions of room temperature, light, 4℃, 45℃, -18℃, and cyclic (-18℃, 4℃, 45℃, with a temperature change every 24 hours), obvious stratification phenomenon was observed.

[0059] Analysis of the stability test results of Example 1 and Comparative Examples 6-9 shows that the particle size of the β-nicotinamide mononucleotide powder and the water-soluble active ingredient powder (adenosine powder) also affects the stability of the NMN oil suspension. The NMN oil suspension prepared using β-nicotinamide mononucleotide powder with a particle size between 400 and 600 nm and adenosine powder with a particle size between 800 and 900 nm as raw materials has better stability. Beyond this particle size range, the NMN oil suspension shows stratification after being placed for 1 month.

[0060] Experimental Example 2: Determination of NMN Residue in Oil Suspension Samples under Room Temperature, Light Irradiation, and High Temperature

[0061] Stability studies were conducted simultaneously on 5% NMN aqueous solution, oil suspensions of Examples 1-14, and Comparative Examples 1-9 at room temperature (25°C), under light irradiation (light intensity 4500 Lx ± 500 Lx, 24-hour continuous light irradiation), and at 45°C for 3 weeks. The NMN residue rate in the oil suspensions was measured after 3 weeks. The NMN residue rate was calculated using the following formula:

[0062] NMN residual rate = (Actual NMN residual amount / Initial NMN content) × 100%

[0063] The method for determining the content of β-nicotinamide mononucleotide (NMN) is as follows:

[0064] a. Plotting the standard curve for β-nicotinamide mononucleotide (NMN)

[0065] Accurately weigh 20.00 mg of β-nicotinamide mononucleotide (NMN) standard, place it in a 10 mL volumetric flask, add water and sonicate for 5 min to dissolve, then dilute to the mark and shake well to obtain a linear stock solution (2.00 mg / mL). -1 The linear stock solution was diluted with water to prepare a series of standard solutions with different mass concentration gradients (0.05, 0.1, 0.2, 0.5, 1.0, and 2.0 mg / mL). -1 After filtering through a 0.22 μm filter membrane, the sample was analyzed by HPLC, and the peak area was recorded. A plot of peak area (Y, mV·s) versus β-nicotinamide mononucleotide (NMN) concentration (ρ, mg·mL) was plotted. -1 The standard curve of the curve was obtained, the regression coefficient was calculated, and the linear equation was obtained by fitting: Y = 2053862.12ρ + 5367.11, r = 1, which showed good linearity.

[0066] The HPLC detection conditions were as follows: YMCC18 column (4.6 mm × 25 mm, 5 μm); flow rate 1.0 mL / min. -1 The column temperature was 40℃; the detection wavelength was 254nm; the injection volume was 20μL; the mobile phase consisted of 50mM KH2PO4 and acetonitrile (95:5, v / v); and the detector was an ultraviolet absorption detector.

[0067] b. Determination of β-nicotinamide mononucleotide (NMN) content in oil suspension samples

[0068] 1 mL of the oil suspension sample after 3 weeks of investigation was accurately transferred into a 10 mL volumetric flask, dissolved by sonication for 5 min, diluted to the mark, and shaken well to obtain the sample solution to be tested. After filtration through a 0.22 μm filter membrane, HPLC detection was performed, the peak area was recorded, the actual residual amount of NMN was calculated according to the standard curve, and the NMN residual rate was calculated. The results are shown in Table 4.

[0069] Table 4. NMN residual rate (%) of oil suspension samples after 3 weeks of storage

[0070]

[0071]

[0072] The results above show that encapsulating β-nicotinamide mononucleotide (NMN) powder in an oil phase is more conducive to the stability of NMN than dissolving it in an aqueous phase. This indicates that encapsulating NMN in an oil phase to form an NMN oil suspension is beneficial to the stability of NMN, thereby maximizing the shelf life of the product.

[0073] Application Examples 1-14:

[0074] The emulsions used in Examples 1-14 are emulsions containing the NMN oil suspensions of Examples 1-14, and the components of the emulsions are as follows:

[0075] Phase A (parts by weight): 1 part A165 (emulsifier), 3 parts octanoic acid / capric triglyceride (GTCC), 3 parts white oil, 0.5 parts cetearyl alcohol, 5 parts silicone oil, and 5 parts NMN oil suspension from Examples 1-14;

[0076] Phase B (parts by weight): 4 parts glycerin, 4 parts butylene glycol, 0.4 parts EMT-10 (thickener), 0.1 parts xanthan gum, 0.4 parts succinate, 0.4 parts hexanediol, 0.2 parts soybean lecithin, and deionized water to make up to 100 parts based on the components of Phase A and Phase B.

[0077] The preparation steps of the above emulsion are as follows:

[0078] 1) First disperse EMT-10, xanthan gum and soybean lecithin into glycerol and butylene glycol, then add succinate, hexanediol and deionized water, heat and stir in an 80°C water bath to fully dissolve them, and obtain phase B;

[0079] 2) A165, GTCC, white oil, cetearyl alcohol, silicone oil, and the NMN oil suspension from Examples 1-14 were heated and melted in a water bath at 83°C to obtain phase A;

[0080] 3) Heat phase A and phase B in a water bath at 83°C. When the two phases are at the same temperature, slowly add phase A to phase B while stirring phase B at 500 rpm. Then start homogenization for 3 min at a homogenization rate of 6000 rpm.

[0081] 4) After homogenization, cool the emulsion at room temperature while stirring at 430 rpm.

[0082] 5) When the temperature drops to 38°C, let it stand and cool to room temperature to obtain an emulsion containing NMN oil suspension.

[0083] Application Example 15:

[0084] The essence of Application Example 15 is a two-component essence containing the NMN oil suspension of Example 1. The formulation of the two-component essence is as follows:

[0085] Formula 1: 3 parts of the NMN oil suspension from Example 1;

[0086] Formula 2: 0.05 parts EDTA-2Na, 4 parts glycerin, 3 parts butylene glycol, 0.1 parts xanthan gum, 0.05 parts hexanediol, 0.5 parts copper sulfate, 0.05 parts sodium hyaluronate (1.2 million), and 92.25 parts deionized water.

[0087] The preparation method of the above-mentioned dual-dosage form essence is as follows:

[0088] (1) Add EDTA-2Na, glycerol, butylene glycol, xanthan gum, sodium hyaluronate (1.2 million), and deionized water to a beaker, heat to 80°C, stir until completely dissolved, and then cool down.

[0089] (2) When the temperature drops to 65℃, add hexanediol and copper sulfate, keep warm and stir for 10 minutes, then stir and cool down.

[0090] (3) When the temperature drops below 45℃, stop stirring to obtain formula 2;

[0091] (4) Formula 1 and Formula 2 are respectively packaged in specific two-agent packaging materials to obtain a two-agent essence.

[0092] Comparative application examples 1-11:

[0093] An emulsion, wherein the emulsions of comparative application examples 1 to 9 each contain the oil suspension of comparative examples 1 to 9, the emulsion of comparative application example 10 is a blank emulsion in which no oil suspension is added but an equal amount of pure water is used instead, and the emulsion of comparative application example 11 is an emulsion in which no oil suspension is added but an equal amount of 5% NMN aqueous solution is used instead.

[0094] The preparation method of the above emulsion is the same as the preparation steps of the emulsions in Application Examples 1 to 14.

[0095] Experimental Example 3: Determination of NMN Leakage Rate of Emulsion Samples under Room Temperature, Light Irradiation, and High Temperature

[0096] Emulsion samples from Application Examples 1-4 and Comparative Application Examples 1-11 were simultaneously subjected to stability tests at room temperature (25°C), under light exposure (light intensity 4500Lx±500Lx, 24-hour continuous light exposure), and at 45°C for one month. The NMN leakage rate in the emulsion samples was measured after one month. The NMN leakage rate was calculated using the following formula:

[0097] NMN leakage rate = (Measured amount of NMN in the aqueous phase of the emulsion sample / Initial NMN content) × 100%

[0098] The method for determining the content of β-nicotinamide mononucleotide (NMN) is as follows:

[0099] a. The standard curve for β-nicotinamide mononucleotide (NMN) was plotted in the same manner as in Experiment 2.

[0100] b. Determination of β-nicotinamide mononucleotide (NMN) content in emulsion samples

[0101] Accurately transfer 0.5 mL of the emulsion sample after one month of investigation into a 10 mL volumetric flask, dissolve it in water, dilute to the mark, shake well, and obtain the sample solution to be tested. Perform HPLC detection, record the peak area, calculate the amount of NMN in the aqueous phase of the emulsion sample according to the standard curve, and calculate the NMN leakage rate.

[0102] Specificity assessment: The above steps were performed on the 5% NMN aqueous solution and the emulsion sample of Comparative Application Example 10. The results showed that the 5% NMN aqueous solution had a significant chromatographic peak at 2.4 min to 2.8 min, while the Comparative Application Example 10 had almost no chromatographic peak at 2.4 min to 2.8 min. This indicates that the components in the Comparative Application Example 10 did not interfere with the determination of NMN, and the determination method has good specificity.

[0103] Criterion Determination: During the one-month stability study, unstable NMN oil suspensions in the emulsion samples experienced some leakage. The leaked NMN also underwent some decomposition over time, making it impossible to determine the specific leakage / residual amount. Therefore, we selected a determination method: for emulsions containing NMN oil suspensions, when the NMN leakage rate is greater than 0.01% (when the NMN oil suspension is stable, its effective residual rate is ≥99.99%, and its leakage rate is definitely <0.01%, therefore, a leakage rate greater than 0.01% is selected as the determination indicator), then the NMN oil suspension is considered to have some leakage. The results are shown in Table 5.

[0104] Table 5. NMN leakage of emulsion samples after 1 month of storage

[0105]

[0106]

[0107] As shown in the table above, the stability of the oil suspension has a significant impact on the NMN leakage rate in the emulsion system when applied to it. A stable oil suspension system (the NMN oil suspensions of Examples 1-14) ensures that NMN does not leak from the oil phase of the emulsion system, thus protecting the stability of NMN within the emulsion system. In contrast, unstable oil suspension systems (the NMN oil suspensions of Comparative Examples 1, 3, 5-9) will experience varying degrees of NMN leakage and cannot protect the stability of NMN within the emulsion system. Therefore, storing NMN in a stable oil suspension system allows it to remain stable within the emulsion system, thereby extending its shelf life.

[0108] Experiment Example 4: Experiment to Promote the Proliferation of Human Skin Fibroblasts

[0109] Human skin fibroblasts (HSF cells, purchased from Guangdong Boxi Biotechnology Co., Ltd.) were used as the test model in this experiment. HSF cells were grown at a concentration of 3.5 × 10⁻⁶. 3 Cells were seeded at a rate of [number] cells / well into 96-well plates and incubated in an incubator (37℃, 5% CO2). When the cell deposition rate in the 96-well plates reached 20%–30%, cells were administered to different groups. The sample information for drug administration is shown in Table 6. The drug dosage per well was 200 μL, with three replicates per group. After drug administration, the cells were incubated in an incubator (37℃, 5% CO2) for 24 h. After 24 h, the supernatant was discarded, and MTT working solution (0.5 mg / mL, freshly prepared) was added. The cells were incubated at 37℃ in the dark for 4 h. After incubation, the supernatant was discarded, and 150 μL of DMSO was added to each well. The OD value was read at 490 nm. The relative cell viability enhancement rate was calculated using the following formula:

[0110]

[0111] The test protocol for the human skin fibroblast proliferation experiment is shown in Table 6.

[0112] Table 6. Test protocol for human skin fibroblast proliferation assay

[0113]

[0114] The results of the relative increase in cell viability are shown in the table below:

[0115] Table 7. Relative cell viability enhancement rate

[0116] Serial Number name 24-hour relative vitality (%) SD P-value 1 BC 143.36 6.73％ / 2 PC 254.78 8.21％ 0.000## 3 Application Example 1 405.68 7.29％ 0.001** 4 Application Example 2 296.56 5.69％ 0.003** 5 Application Example 3 266.69 7.68％ 0.000** 6 Application Example 4 250.11 10.27％ 0.015* 7 Application Example 5 404.32 3.26％ 0.001** 8 Application Example 6 407.01 5.99％ 0.003** 9 Application Example 7 405.75 4.86％ 0.004** 10 Application Example 8 403.68 8.47％ 0.001** 11 Application Example 9 405.16 4.89％ 0.002** 12 Application Example 10 402.38 6.79％ 0.004** 13 Application Example 11 406.75 5.89％ 0.001** 14 Application Example 12 405.59 7.27％ 0.000** 15 Application Example 13 403.45 5.33％ 0.001** 16 Application Example 14 404.26 6.19％ 0.003** 17 Comparative Application Example 1 164.35 8.62％ 0.005** 18 Comparative Application Example 2 178.34 4.96％ 0.009** 19 Comparative Application Example 3 258.68 7.29％ 0.21 20 Comparative Application Example 4 288.83 8.39％ 0.011* 21 Comparative Application Example 5 162.39 4.36％ 0.013* 22 Comparative Application Example 6 401.68 8.69％ 0.002** 23 Comparative Application Example 7 397.25 11.36％ 0.015* 24 Comparative Application Example 8 405.21 4.89％ 0.007** 25 Comparative Application Example 9 399.99 7.36％ 0.024* 26 Comparative Application Example 10 145.56 2.12％ 0.11 27 Comparative Application Example 11 165.49 0.0547 0.013*

[0117] Note: ## indicates p < 0.01 compared to the blank control; ** indicates p < 0.01 compared to the negative control; * indicates 0.01 < p < 0.05 compared to the negative control.

[0118] As shown in the table above, compared with the BC group, the PC group significantly promoted the proliferation of HSF cells 24 hours after administration of 10% NBS. Analysis of the relative cell viability enhancement rates of Application Example 1 and the comparative application examples 1, 2, 3, and 4 shows that when β-nicotinamide mononucleotide and adenosine are used in combination, their effect on HSF cell proliferation is significantly better than that of using either alone. This indicates that the NMN oil suspension of the present invention has a good HSF cell proliferation promoting effect and good anti-aging efficacy, and that the combination of β-nicotinamide mononucleotide and adenosine has a certain synergistic effect.

Claims

1. An NMN oil suspension composition, characterized in that, The NMN oil suspension composition comprises, in parts by weight: 0.5-6 parts of β-nicotinamide mononucleotide, 0.2-3 parts of water-soluble active ingredient, and an oil-phase emollient to make up to 100 parts; the NMN oil suspension is made by compounding β-nicotinamide mononucleotide and water-soluble active ingredient and encapsulating them with an oil-phase emollient, wherein the water-soluble active ingredient is adenosine.

2. The NMN oil suspension composition according to claim 1, characterized in that, The NMN oil suspension composition comprises, in parts by weight: 5 parts β-nicotinamide mononucleotide, 1.8 parts water-soluble active ingredient, and oil phase emollient to make up to 100 parts.

3. The NMN oil suspension composition according to claim 1, characterized in that, The β-nicotinamide mononucleotide is in powder form with a particle size between 400 and 600 nm and a regular spherical microstructure.

4. The NMN oil suspension composition according to claim 1, characterized in that, The water-soluble active ingredient is in powder form, with a particle size between 800 and 900 nm and a regular spherical microstructure.

5. The NMN oil suspension composition as described in claim 1, characterized in that, The oil-phase emollient is one or more of the following: isopropyl palmitate, isopropyl myristate, isooctyl palmitate, dioctyl carbonate, caprylic / capric triglyceride, squalane, meadowfoam seed oil, sunflower seed oil, vitamin E acetate, hydrogenated polyisobutylene, isononyl isononanoate, C12-15 alcohol benzoate, marula oil, emu oil, sweet almond oil, C9-12 alkyl, coconut oil-caprylic / capric ester, polydimethylsiloxane, plant squalane, isooctyl palmitate, and isopropyl myristate.

6. A method for preparing the NMN oil suspension according to claim 1, characterized in that, Includes the following steps: S1. Under normal temperature conditions, β-nicotinamide mononucleotide and water-soluble active ingredient are mechanically ball-milled to obtain β-nicotinamide mononucleotide powder and water-soluble active ingredient powder that meet the particle size requirements. S2. Under normal temperature conditions, β-nicotinamide mononucleotide powder is added to 1 / 2 to 3 / 4 of the oil phase emollient and dispersed by high-speed shearing, and then homogenized under high pressure to obtain oil 1; S3. Under normal temperature conditions, the water-soluble active ingredient powder is added to the remaining oil phase emollient and dispersed by high-speed shearing, and then homogenized under high pressure to obtain oil 2. S4. Mix oil 1 and oil 2, stir and disperse evenly at room temperature, and then homogenize under high pressure to obtain NMN oil suspension.

7. The preparation method according to claim 6, characterized in that, In step S2, the high-pressure homogenization process involves a homogenization rate of 6000~8000 rpm, a homogenization time of 8~12 min, and a homogenization pressure of 1.4~1.6 MPa; the process is repeated 3~6 times.

8. The preparation method according to claim 6, characterized in that, In step S2, the high-pressure homogenization process involves a homogenization rate of 7500 rpm, a homogenization time of 10 min, and a homogenization pressure of 1.5 MPa; the process is repeated 4 times.

9. The preparation method according to claim 6, characterized in that, In step S3, the high-pressure homogenization process involves a homogenization rate of 4000~6000 rpm, a homogenization time of 6~8 min, and a homogenization pressure of 1.0~1.2 MPa; the process is repeated 2~4 times.

10. The preparation method according to claim 6, characterized in that, In step S3, the high-pressure homogenization process involves a homogenization rate of 5000 rpm, a homogenization time of 7 min, and a homogenization pressure of 1.1 MPa; the process is repeated 3 times.

11. The preparation method according to claim 6, characterized in that, In step S4, the high-pressure homogenization process involves a homogenization rate of 4000~6000 rpm, a homogenization time of 4~6 min, and a homogenization pressure of 1.2~1.4 MPa; the process is repeated 2~4 times.

12. The preparation method according to claim 6, characterized in that, In step S4, the high-pressure homogenization process involves a homogenization rate of 5500 rpm, a homogenization time of 5 min, and a homogenization pressure of 1.3 MPa; the process is repeated twice.

13. A cosmetic product with anti-aging effects, characterized in that, The cosmetic product comprises the NMN oil suspension as described in claim 1 or 6.

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