A method for improving the non-invasive anti-aging activity of fine line suppression collagen loss by complexing

Abstract

The present application relates to the field of cosmetic active compound, and discloses a non-invasive anti-aging active compound preparation method for improving fine line inhibition and collagen loss, which comprises a collagen degradation inhibition compound phase, a stress buffering isolation phase and a collagen signal maintenance phase; wherein the collagen degradation inhibition compound phase comprises hydroxytyrosol, zinc pyrrolidone carboxylic acid, 1,3-propanediol and deionized water, the stress buffering isolation phase comprises ikaros, ergothioneine, trehalose, beta-glucan, pullulan and deionized water, and the collagen signal maintenance phase comprises N-acetyl hydroxyproline, hexapeptide-9, palmitoyl tripeptide-5, 1,3-propanediol and deionized water; in the preparation, the collagen degradation inhibition compound phase is prepared first, then the stress buffering isolation phase is added, and finally the collagen signal maintenance phase is added under the condition that the temperature is not higher than 35 DEG C, and the pH of the final system is controlled to be 5.3-5.8; the present application can improve the compatibility stability and preparation repeatability of multiple types of anti-aging active compounds in weak acid aqueous phase, and is suitable for external skin care preparations.

Description

Technical Field

[0001] This invention relates to the field of cosmetic active ingredient compounding, specifically to a non-invasive anti-aging active ingredient compounding method for improving fine lines and inhibiting collagen loss. Background Technology

[0002] As topical skincare products evolve towards anti-wrinkle, firming, and barrier repair, there is a growing trend of active ingredient formulations focused on improving fine lines and inhibiting collagen loss. Current anti-aging skincare products typically use retinol, retinyl esters, hydroxypropyl tetrahydropyranotriols, peptides, hyaluronic acid, collagen, or plant antioxidants as their main active ingredients. These ingredients improve the appearance of aging skin by promoting keratinocyte renewal, increasing skin hydration, replenishing exogenous collagen, or regulating collagen-related expression. While these approaches can improve dryness lines, fine lines, and decreased skin elasticity to some extent, the stability, irritation, order of addition, and system compatibility of different active ingredients vary considerably. Simply adding multiple anti-aging ingredients directly in parallel to the same aqueous phase or emulsion system can easily lead to problems such as excessively high local concentrations, system color changes, fluctuations in active ingredient content, and prolonged direct contact between peptide components and polyphenolic components due to heat.

[0003] In existing technologies, hydroxytyrosol, ectoine, ergothioneine, polysaccharides, and peptides can all be used as topical skincare active ingredients. However, most solutions focus on the synergistic effects of single active ingredients or directly mix functional ingredients such as antioxidants, moisturizers, collagen promoters, and soothing repair agents to form a composition. While this type of compounding method is simple to prepare, it does not adequately consider the sequential relationships between collagen degradation, oxidative stress, glycation stress, and the maintenance of collagen-related signals involved in the formation of fine lines. It also fails to fully address the compatibility and stability issues of polyphenolic active ingredients, zinc salts, polysaccharide components, and peptide components in the same weakly acidic aqueous system. In particular, polyphenolic substances such as hydroxytyrosol are easily affected by pH, temperature, light, and the state of metal ions in aqueous systems, resulting in color or content fluctuations. Furthermore, prolonged treatment of peptide active ingredients at high temperatures or in complex mixed environments may also affect the stability and batch consistency of the final compounded system.

[0004] Therefore, there is still a need for a non-invasive method for compounding anti-aging active ingredients suitable for topical skincare formulations. This method should not simply involve mixing antioxidants, moisturizers, polysaccharides, and peptides all at once. Instead, it should establish a phase-separated preparation and sequential compounding approach based on the compatibility and addition conditions of different active ingredients. This allows collagen degradation inhibitory active ingredients, stress buffering active ingredients, and collagen signaling maintenance active ingredients to be pre-dispersed or pre-compounded in suitable aqueous environments before being added to the final system. This approach improves the stability of the compounded solution, pH compatibility, and compatibility when subsequently added to skincare bases such as serums, lotions, creams, or masks. This provides a technical solution that differs from the conventional direct compounding of active ingredients for addressing skincare needs related to improving fine lines and inhibiting collagen loss. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a non-invasive method for compounding anti-aging active ingredients to improve fine lines and inhibit collagen loss, thereby solving the technical problems existing in the prior art.

[0006] The above-mentioned technical objective of the present invention is achieved through the following technical solution: A non-invasive anti-aging active ingredient compound solution that improves fine lines and inhibits collagen loss includes the following steps: The non-invasive anti-aging active ingredient compound solution is formed by first adding the stress buffer isolation phase to the collagen degradation inhibitory compound phase, and then adding the collagen signal maintenance phase. The collagen degradation inhibitor complex consists of hydroxytyrosol, zinc pyrrolidone carboxylate, 1,3-propanediol, and deionized water; The stress buffering phase includes ectoine, ergothioneine, trehalose, β-glucan, pullulan, and deionized water; The collagen signal maintenance phase includes N-acetylhydroxyproline, hexapeptide-9, palmitoyl tripeptide-5, 1,3-propanediol, and deionized water; Based on the total mass of the non-invasive anti-aging active ingredient compound solution, the non-invasive anti-aging active ingredient compound solution comprises the following components by mass percentage: hydroxytyrosol 0.10–0.60%, zinc pyrrolidone carboxylate 0.07–0.66%, ectoine 1.50–3.00%, ergothioneine 0.03–0.07%, trehalose 2.00–8.00%, β-glucan 0.10–0.40%, pullulan 0.04–0.20%, N-acetylated hydroxyproline 0.60–1.20%, hexapeptide-9 0.006–0.024%, palmitoyl tripeptide-5 0.0024–0.012%, 1,3-propanediol 6.00–18.00%, pH adjuster 0.01–0.30%, with the balance being deionized water.

[0007] Preferably, the mass ratio of hydroxytyrosol to zinc pyrrolidone carboxylate is 1:0.7 to 1.1; The mass ratio of ectoine to ergothioneine is 30–70:1; The mass ratio of the β-glucan to pullulan is 1:0.2 to 1.5.

[0008] Preferably, the mass ratio of N-acetylhydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 is 100:0.6-2.5:0.2-1.0; The mass ratio of the collagen degradation inhibitory complex phase, the stress buffer isolation phase, and the collagen signal maintenance phase is 1:2.0-5.0:0.3-0.8.

[0009] Preferably, the pH adjuster is composed of citric acid and sodium citrate, and the pH of the non-invasive anti-aging active ingredient compound solution is 5.3 to 5.8; The non-invasive anti-aging active ingredient compound solution is filtered through a 0.45 μm pore size filter membrane and then placed into an opaque container.

[0010] Preferably, the non-invasive anti-aging active ingredient compound solution does not contain retinol active ingredients, hydroxypinazone retinate, hydroxypropyl tetrahydropyranotriol, acetyl hexapeptide-8 and exogenous collagen active ingredients; The retinol-based active ingredients include retinol, retinaldehyde, retinyl palmitate, and retinyl acetate; The exogenous collagen-like active ingredients include hydrolyzed collagen, soluble collagen, and recombinant collagen.

[0011] A method for preparing a non-invasive anti-aging active ingredient compound solution that improves fine lines and inhibits collagen loss includes the following steps: S1: Preparation of collagen degradation inhibitory complex phase: Hydroxytyrosol was added to an aqueous phase containing 1,3-propanediol and stirred to dissolve at 35-45℃. Then, zinc pyrrolidone carboxylate was added, and the pH of the system was adjusted to 5.2-5.6 using a pH adjuster. The mixture was stirred for 30-60 min to obtain the collagen degradation inhibitory complex phase. S2: Preparation of stress buffer isolation phase: Add ectoine, trehalose, ergothioneine, β-glucan and pullulan to the aqueous phase, and stir at 25-35℃ until the system is homogeneous to obtain the stress buffer isolation phase; S3: Preparation of collagen signal maintenance phase: Dissolve N-acetylhydroxyproline in aqueous phase, cool to no higher than 35℃, add hexapeptide-9 and palmitoyl tripeptide-5, stir for 10-25 min to obtain collagen signal maintenance phase; S4: Sequential compounding: First, add the stress buffer isolation phase to the collagen degradation inhibition compound phase and stir for 20-40 minutes. Then, add the collagen signal maintenance phase to the mixed system and control the final system pH to 5.3-5.8 to obtain a non-invasive anti-aging active ingredient compound solution.

[0012] Preferably, the aqueous phase containing 1,3-propanediol is composed of 1,3-propanediol and deionized water, and the mass percentage of 1,3-propanediol in the collagen degradation inhibitory complex phase is 10.00-20.00%. After the hydroxytyrosol is added to the aqueous phase containing 1,3-propanediol, it is stirred at a speed of 300-600 r / min for 10-20 min, and then zinc pyrrolidone carboxylate is added.

[0013] Preferably, the pH adjuster comprises an aqueous solution of citric acid and an aqueous solution of sodium citrate, wherein the concentration of the aqueous solution of citric acid is 0.05–0.20 mol / L and the concentration of the aqueous solution of sodium citrate is 0.05–0.20 mol / L. After adjusting the pH of the system, continue stirring the system at 35–45°C for 30–60 min.

[0014] Preferably, in step S2, trehalose, ectoine, and ergothioneine are first added to deionized water and stirred to dissolve, and then β-glucan and pullulan are added and stirred to disperse. The stirring speed after adding the β-glucan and pullulan is 400-800 r / min, and the stirring time is 20-50 min.

[0015] Preferably, the mass ratio of the collagen degradation inhibitory complex phase, the stress buffering and isolating phase, and the collagen signal maintenance phase is 1:2.0-5.0:0.3-0.8; In step S4, when the stress buffer isolation phase is added to the collagen degradation inhibitory complex phase, the addition rate is 2.00-8.00% of the mass of the collagen degradation inhibitory complex phase per minute, and the stirring speed during the addition is 300-600 r / min. When the collagen signal maintenance phase is added to the mixing system, the temperature of the mixing system is 25-35℃, the addition rate is 0.50-3.00% of the mass of the mixing system per minute, and the stirring speed during the addition is 200-500 r / min; After all the collagen signal maintenance phase is added, continue stirring for 10–30 minutes.

[0016] In summary, the present invention has the following main beneficial effects: By first preparing a collagen degradation inhibitory complex phase using hydroxytyrosol and zinc pyrrolidone carboxylate in a weakly acidic aqueous phase containing 1,3-propanediol, and controlling the pH of this complex phase at 5.2–5.6, hydroxytyrosol and zinc pyrrolidone carboxylate are pre-dispersed and compatibly compounded before entering the final system. This reduces color changes, precipitation, or content fluctuations of hydroxytyrosol in the aqueous system due to excessively high local concentrations, pH fluctuations, or direct exposure to complex matrices. Simultaneously, this complex phase is independent of retinols, hydroxypinazone retinate, hydroxypropyl tetrahydropyranotriol, or exogenous collagen active ingredients, distinguishing it from existing stimulating renewal pathways and exogenous collagen supplementation pathways. This makes the complex more suitable as a mild anti-aging active ingredient additive in topical skincare formulations.

[0017] By preparing a stress-buffered isolating phase using ectoine, ergothioneine, trehalose, β-glucan, and pullulan, and then adding this stress-buffered isolating phase to a collagen degradation-inhibiting complex phase followed by a collagen signal-maintaining phase, a dispersion buffer environment is established between the polyphenolic actives, zinc salts, and peptide actives. Compared to mixing antioxidants, humectants, polysaccharides, and peptides all at once, the sequential compounding process of this application reduces the risk of localized high-concentration contact between the actives upon addition. This allows β-glucan and pullulan to first form a uniformly dispersed aqueous phase in the system, which facilitates the subsequent entry of the collagen signal-maintaining phase into the system under lower shear and lower temperature conditions, improving the repeatability and system compatibility of the compound preparation process.

[0018] By preparing a collagen signal maintenance phase containing N-acetylhydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 at temperatures not exceeding 35°C, and controlling the mass ratio of the collagen degradation inhibitory complex phase, stress buffering isolation phase, and collagen signal maintenance phase to 1:2.0–5.0:0.3–0.8, the collagen-related active ingredients are compounded under low-temperature, weakly acidic, and uniformly dispersed conditions. This treatment method avoids prolonged heating or direct involvement in complex mixing of peptide components during the pre-compounding stage of hydroxytyrosol and zinc pyrrolidone carboxylate, thus providing a better process foundation in terms of appearance stability, pH compatibility, filtration operability, and compatibility when subsequently added to serum, emulsion, cream, or mask base. Attached Figure Description

[0019] Figure 1 This is a flowchart of the method of the present invention. Detailed Implementation

[0020] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] refer to Figure 1 A non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss includes a collagen degradation inhibitory compound phase, a stress buffer isolation phase and a collagen signal maintenance phase. The non-invasive anti-aging active ingredient compound solution is formed by first adding the stress buffer isolation phase to the collagen degradation inhibitory compound phase and then adding the collagen signal maintenance phase. The collagen degradation inhibitor complex consists of hydroxytyrosol, zinc pyrrolidone carboxylate, 1,3-propanediol, and deionized water; The stress buffering phase includes ectoine, ergothioneine, trehalose, β-glucan, pullulan, and deionized water; The collagen signal maintenance phase includes N-acetylhydroxyproline, hexapeptide-9, palmitoyl tripeptide-5, 1,3-propanediol, and deionized water; Based on the total mass of the non-invasive anti-aging active ingredient compound solution, the non-invasive anti-aging active ingredient compound solution comprises the following components by mass percentage: hydroxytyrosol 0.10–0.60%, zinc pyrrolidone carboxylate 0.07–0.66%, ectoine 1.50–3.00%, ergothioneine 0.03–0.07%, trehalose 2.00–8.00%, β-glucan 0.10–0.40%, pullulan 0.04–0.20%, N-acetylated hydroxyproline 0.60–1.20%, hexapeptide-9 0.006–0.024%, palmitoyl tripeptide-5 0.0024–0.012%, 1,3-propanediol 6.00–18.00%, pH adjuster 0.01–0.30%, with the balance being deionized water.

[0022] The non-invasive anti-aging active ingredient compound solution described in this application is a topical skincare active ingredient compound solution and is not intended for injection, skin breakage, wound treatment, or medical treatment. The improvement of fine lines and inhibition of collagen loss described in this application refers to the compound solution being formulated as an active ingredient additive in topical skincare preparations, focusing on collagen degradation-related stress, active ingredient stability, and the compatibility of collagen-related active ingredients. It should not be construed as a medical application or a treatment of collagen metabolism in disease states.

[0023] The hydroxytyrosol, zinc pyrrolidone carboxylate, ectoine, ergothioneine, trehalose, β-glucan, pullulan, N-acetylhydroxyproline, hexapeptide-9, palmitoyl tripeptide-5, 1,3-propanediol, citric acid, and sodium citrate used in the embodiments of this application can all be cosmetic-grade or topical skincare raw material grade products. The purity, moisture content, heavy metal content, and microbial limits of each raw material can be confirmed based on the quality inspection reports, enterprise standards, or cosmetic raw material safety data provided by the supplier. All mass percentages in the embodiments are based on the total mass of the final non-invasive anti-aging active ingredient compound solution.

[0024] In this application, the collagen degradation inhibitory complex phase comprises hydroxytyrosol, zinc pyrrolidone carboxylate, 1,3-propanediol, and deionized water; the stress buffering phase comprises ectoine, ergothioneine, trehalose, β-glucan, pullulan, and deionized water; and the collagen signal maintenance phase comprises N-acetylhydroxyproline, hexapeptide-9, palmitoyl tripeptide-5, 1,3-propanediol, and deionized water. The collagen degradation inhibitory complex phase is used to pre-contact hydroxytyrosol and zinc pyrrolidone carboxylate in a weakly acidic aqueous phase containing 1,3-propanediol; the stress buffering phase is used to form a polysaccharide-containing aqueous dispersion during subsequent compounding; and the collagen signal maintenance phase is used to introduce N-acetylhydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 under low-temperature conditions.

[0025] In this application, the pH of the collagen degradation inhibitor complex phase is controlled at 5.2–5.6. This range is determined based on the dispersion stability of hydroxytyrosol in an aqueous phase containing 1,3-propanediol, the compatibility of zinc pyrrolidone carboxylate in a weakly acidic aqueous phase, and the subsequent peptide phase addition conditions. When the pH is below 5.2, the system compatibility decreases after the subsequent collagen signal maintenance phase is added; when the pH is above 5.6, the risk of color change of hydroxytyrosol during storage increases. The final non-invasive anti-aging active ingredient complex solution has a pH controlled at 5.3–5.8, which matches the weakly acidic usage environment of the topical skin care aqueous phase system and can also take into account the compatibility after the three-phase complex is formulated.

[0026] Temperature S1 is controlled at 35–45°C to accelerate the dissolution and dispersion of hydroxytyrosol in an aqueous phase containing 1,3-propanediol. Below 35°C, the time required for hydroxytyrosol and zinc pyrrolidone carboxylate to reach a homogeneous state is prolonged; above 45°C, the risk of color change in hydroxytyrosol during subsequent storage evaluation increases. Temperatures S2 and S3 are controlled at 25–35°C to meet the aqueous dispersion requirements of ectoine, trehalose, ergothioneine, polysaccharides, and peptides; S3 is limited to no higher than 35°C to avoid prolonged treatment of hexapeptide-9 and palmitoyl tripeptide-5 at higher temperatures.

[0027] The mass ratio of hydroxytyrosol to zinc pyrrolidone carboxylate was controlled at 1:0.7–1.1. When the zinc pyrrolidone carboxylate was below this range, its contribution to the stability of the aqueous phase of hydroxytyrosol in the collagen degradation inhibitory complex was insufficient; when it was above this range, the risk of visible precipitation after centrifugation increased. The mass ratio of ectoine to ergothioneine was controlled at 30–70:1, based on the fact that ergothioneine has a low amount of active ingredient; too high a ratio would increase system color and cost fluctuations, while too low a ratio would not be sufficient to form a stable stress-buffered isolating phase. The mass ratio of β-glucan to pullulan was controlled at 1:0.2–1.5, based on a comprehensive selection of polysaccharide dispersibility, system viscosity, and operability at 0.45m filtration; when pullulan was below this range, the dispersion framework of the stress-buffered isolating phase was insufficient; when pullulan was above this range, system viscosity and filtration resistance increased significantly.

[0028] The mass ratio of N-acetylhydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 is controlled at 100:0.6–2.5:0.2–1.0. N-acetylhydroxyproline serves as the main water-soluble component in the collagen signal maintenance phase, while hexapeptide-9 and palmitoyl tripeptide-5 are low-addition peptide components. This ratio balances aqueous dissolution, low-temperature addition, and final solution concentration control. The mass ratio of the collagen degradation inhibitory phase, stress buffer phase, and collagen signal maintenance phase is controlled at 1:2.0–5.0:0.3–0.8. If the stress buffer phase ratio is too low, the isolation and dispersion effect is insufficient; if the stress buffer phase ratio is too high, it will dilute the active ingredient concentrations in both the collagen degradation inhibitory phase and the collagen signal maintenance phase; if the collagen signal maintenance phase ratio is too low, it will be difficult to uniformly add the peptide active ingredients; if the collagen signal maintenance phase ratio is too high, it will increase the fluctuation of the aqueous phase ratio in the final system.

[0029] This application uses citric acid and sodium citrate as pH adjusters. Citric acid and sodium citrate can form a weakly acidic buffer system, suitable for controlling the collagen degradation inhibitory complex phase and the final complex solution within the ranges of 5.2–5.6 and 5.3–5.8, respectively, avoiding local pH abrupt changes when using strong acids or bases for adjustment. The mass percentage of the pH adjuster is 0.01–0.30%. This amount is not added as a fixed active ingredient, but rather is the adjusted process dosage based on the initial pH and target pH range of each batch of raw materials. In the examples, the converted mass of the pH adjuster is based on the actual mass of citric acid and sodium citrate added.

[0030] The non-invasive anti-aging active ingredient compound solution described in this application does not contain retinol-based active ingredients, hydroxypinazone retinoate, hydroxypropyl tetrahydropyranotriol, acetyl hexapeptide-8, or exogenous collagen-based active ingredients. The retinol-based active ingredients include retinol, retinaldehyde, retinyl palmitate, and retinyl acetate; the exogenous collagen-based active ingredients include hydrolyzed collagen, soluble collagen, and recombinant collagen. This limitation means that the above-mentioned substances are not added as active ingredient raw materials to the formulation system when preparing the non-invasive anti-aging active ingredient compound solution. The raw material lists and preparation steps of Examples 1-4 do not include the above-mentioned excluded active ingredients.

[0031] Example 1 This embodiment prepares 1000g of a non-invasive anti-aging active ingredient compound solution. The amounts of each component are as follows: hydroxytyrosol 3.00g, zinc pyrrolidone carboxylate 2.40g, ectoine 20.00g, ergothioneine 0.40g, trehalose 50.00g, β-glucan 2.00g, pullulan 1.00g, N-acetylhydroxyproline 8.00g, hexapeptide-90.12g, palmitoyl tripeptide-50.048g, 1,3-propanediol 120.00g, pH adjuster 1.00g, and the balance is deionized water 792.032g. The pH adjuster consists of citric acid and sodium citrate, and the pH is adjusted using 0.10mol / L citric acid aqueous solution and 0.10mol / L sodium citrate aqueous solution, respectively.

[0032] In this embodiment, the masses of the collagen degradation inhibition complex phase, the stress buffer isolation phase, and the collagen signal maintenance phase are 250.00 g, 600.00 g, and 150.00 g, respectively, and the mass ratio of the three is 1:2.4:0.6.

[0033] S1, Preparation of the collagen degradation inhibitory complex phase. 204.00 g of deionized water and 40.00 g of 1,3-propanediol were added to a clean, jacketed mixing tank with a stirrer. Stirring was started and the system was heated to 40°C, with the stirring speed controlled at 450 r / min. 3.00 g of hydroxytyrosol was slowly added to the mixing tank, and stirring was continued for 15 min to disperse and dissolve the hydroxytyrosol. Then, 2.40 g of zinc pyrrolidone carboxylate was added, and stirring was maintained at 40°C. The pH of the system was adjusted to 5.4 using 0.10 mol / L citric acid aqueous solution and 0.10 mol / L sodium citrate aqueous solution, with an equivalent mass of 0.60 g of pH adjuster added. After adjustment, stirring was continued for 45 min to obtain 250.00 g of the collagen degradation inhibitory complex phase.

[0034] S2, Preparation of the stress buffer isolating phase. Add 526.20 g of deionized water to another clean mixing tank, maintain the temperature at 30°C, and start stirring. First, add 50.00 g of trehalose and stir until dissolved; then add 20.00 g of ectoine and 0.40 g of ergothioneine, and continue stirring until the system is homogeneous. Subsequently, add 2.00 g of β-glucan and 1.00 g of pullulan in batches, maintaining a stirring speed of 600 r / min during the addition process. After all the ingredients are added, continue stirring for 35 min. Adjust the pH of the system to 5.5 using a pH adjuster (equivalent to 0.40 g), yielding 600.00 g of the stress buffer isolating phase.

[0035] S3, Preparation of the collagen signal maintenance phase. Add 61.832 g of deionized water and 80.00 g of 1,3-propanediol to the third mixing tank, start stirring, and control the temperature at 30°C. Add 8.00 g of N-acetylhydroxyproline and stir until dissolved; after confirming that the system temperature does not exceed 35°C, add 90.12 g of hexapeptide- and 50.048 g of palmitoyl tripeptide-, and stir for 20 min to obtain 150.00 g of the collagen signal maintenance phase.

[0036] S4, Sequential Compounding. The collagen degradation inhibitory compound phase prepared in S1 was maintained at 30℃ with a stirring speed of 450 r / min. The stress buffer isolation phase prepared in S2 was added to the collagen degradation inhibitory compound phase at a rate of 10 g / min, which is equivalent to 4.00% of the mass of the collagen degradation inhibitory compound phase added per minute. After all the stress buffer isolation phase was added, stirring was continued for 30 min to obtain a first mixed system with a mass of 850.00 g. Subsequently, the collagen signal maintenance phase prepared in S3 was added to the first mixed system at a rate of 10 g / min, which is equivalent to 1.18% of the mass of the first mixed system added per minute. During the addition, the stirring speed was controlled at 350 r / min, and the system temperature was controlled at 30℃. After all the collagen signal maintenance phase was added, stirring was continued for 20 min. The final pH of the system was detected and adjusted to 5.5 to obtain a non-invasive anti-aging active ingredient compound solution.

[0037] S5, Post-processing. The non-invasive anti-aging active ingredient compound solution obtained in S4 was filtered through a 0.45 μm filter membrane, and then placed in an opaque container and allowed to stand at 25°C for 18 hours. After standing, the pH was measured again, and the pH was maintained within the range of 5.3 to 5.8.

[0038] In this embodiment, the mass ratio of hydroxytyrosol to zinc pyrrolidone carboxylate is 1:0.8; the mass ratio of ectoine to ergothioneine is 50:1; the mass ratio of β-glucan to pullulan is 1:0.5; and the mass ratio of N-acetylated hydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 is 100:1.5:0.6.

[0039] Example 2 This embodiment prepares 1000g of non-invasive anti-aging active ingredient compound solution. The amounts of each component are as follows: hydroxytyrosol 1.00g, zinc pyrrolidone carboxylate 0.80g, ectoine 15.00g, ergothioneine 0.30g, trehalose 20.00g, β-glucan 1.00g, pullulan 0.40g, N-acetylhydroxyproline 6.00g, hexapeptide-90.06g, palmitoyl tripeptide-50.024g, 1,3-propanediol 60.00g, pH adjuster 0.50g, and the balance is deionized water 894.916g.

[0040] In this embodiment, the masses of the collagen degradation inhibition complex phase, the stress buffer isolation phase, and the collagen signal maintenance phase are 260.00 g, 620.00 g, and 120.00 g, respectively, and the mass ratio of the three is 1:2.38:0.46.

[0041] S1. Preparation of a collagen degradation inhibitory complex. The collagen degradation inhibitory complex consisted of 260.00 g of deionized water, 30.00 g of 1,3-propanediol, 1.00 g of hydroxytyrosol, 0.80 g of zinc pyrrolidone carboxylate, and 0.30 g of pH adjuster. Deionized water and 1,3-propanediol were added to a mixing tank, the temperature was raised to 35°C, and the mixture was stirred at 300 rpm. After adding hydroxytyrosol, stirring was continued for 20 min, followed by the addition of zinc pyrrolidone carboxylate. The pH of the system was adjusted to 5.2 using 0.10 mol / L citric acid aqueous solution and 0.10 mol / L sodium citrate aqueous solution. After adjustment, stirring was continued for 60 min to obtain the collagen degradation inhibitory complex. The mass percentage of 1,3-propanediol in the collagen degradation inhibitory complex was 11.54%.

[0042] S2, Preparation of the stress buffer isolating phase. The stress buffer isolating phase, with a volume of 620.00 g, consists of 583.10 g deionized water, 15.00 g ectoine, 0.30 g ergothioneine, 20.00 g trehalose, 1.00 g β-glucan, 0.40 g pullulan, and 0.20 g pH adjuster. Deionized water was added to a mixing tank, and the temperature was controlled at 25°C. Trehalose, ectoine, and ergothioneine were added first and stirred until dissolved. Then, β-glucan and pullulan were added. After adding β-glucan and pullulan, the stirring speed was 400 r / min, and the stirring time was 50 min, yielding the stress buffer isolating phase.

[0043] S3, Preparation of the collagen signal maintenance phase. The collagen signal maintenance phase consisted of 120.00 g of deionized water (83.916 g), 1,3-propanediol (30.00 g), N-acetylhydroxyproline (6.00 g), hexapeptide-9 (0.06 g), and palmitoyl tripeptide-5 (0.024 g). Deionized water and 1,3-propanediol were added to a mixing tank, and the temperature was controlled at 25°C. N-acetylhydroxyproline was added and stirred until dissolved. After confirming that the system temperature did not exceed 35°C, hexapeptide-9 and palmitoyl tripeptide-5 were added, and stirring was continued for 25 min to obtain the collagen signal maintenance phase.

[0044] S4, Sequential Compounding. First, the stress buffer isolation phase is added to the collagen degradation inhibitor compound phase at a rate of 2.00% of the collagen degradation inhibitor compound phase mass per minute, with a stirring speed of 300 r / min during addition. After all the stress buffer isolation phase is added, stirring continues for 40 min. Then, the collagen signal maintenance phase is added at a rate of 0.50% of the mixed system mass per minute, with a stirring speed of 200 r / min during addition, and the system temperature is controlled at 25℃. After all the collagen signal maintenance phase is added, stirring continues for 30 min, and the final system pH is controlled to 5.3, yielding a non-invasive anti-aging active ingredient compound solution. The resulting compound solution is filtered through a 0.45 μm pore size filter membrane and then placed into an opaque container.

[0045] In this embodiment, the mass ratio of hydroxytyrosol to zinc pyrrolidone carboxylate is 1:0.8; the mass ratio of ectoine to ergothioneine is 50:1; the mass ratio of β-glucan to pullulan is 1:0.4; and the mass ratio of N-acetylated hydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 is 100:1.0:0.4.

[0046] Example 3 This embodiment prepares 1000g of non-invasive anti-aging active ingredient compound solution. The amounts of each component are as follows: hydroxytyrosol 6.00g, zinc pyrrolidone carboxylate 6.60g, ectoine 30.00g, ergothioneine 0.60g, trehalose 80.00g, β-glucan 4.00g, pullulan 2.00g, N-acetylhydroxyproline 12.00g, hexapeptide-90.24g, palmitoyl tripeptide-50.096g, 1,3-propanediol 120.00g, pH adjuster 3.00g, and the balance is deionized water 735.464g.

[0047] In this embodiment, the masses of the collagen degradation inhibition complex phase, the stress buffer isolation phase, and the collagen signal maintenance phase are 200.00 g, 650.00 g, and 150.00 g, respectively, and the mass ratio of the three is 1:3.25:0.75.

[0048] S1. Preparation of a collagen degradation inhibitory complex. The collagen degradation inhibitory complex consisted of 200.00 g of deionized water, 40.00 g of 1,3-propanediol, 6.00 g of hydroxytyrosol, 6.60 g of zinc pyrrolidone carboxylate, and 3.00 g of pH adjuster. Deionized water and 1,3-propanediol were added to a mixing tank, the temperature was raised to 45°C, and the mixture was stirred at 600 rpm. After adding hydroxytyrosol, stirring was continued for 10 min, followed by the addition of zinc pyrrolidone carboxylate. The pH of the system was adjusted to 5.6 using 0.10 mol / L citric acid aqueous solution and 0.10 mol / L sodium citrate aqueous solution. After adjustment, stirring was continued for 30 min to obtain the collagen degradation inhibitory complex. The mass percentage of 1,3-propanediol in the collagen degradation inhibitory complex was 20.00%.

[0049] S2, Preparation of the stress buffer isolating phase. The stress buffer isolating phase consisted of 650.00 g of deionized water, 30.00 g of ectoine, 0.60 g of ergothioneine, 80.00 g of trehalose, 4.00 g of β-glucan, and 2.00 g of pullulan. Deionized water was added to a mixing tank, and the temperature was controlled at 35℃. Trehalose, ectoine, and ergothioneine were added first and stirred until dissolved. Then, β-glucan and pullulan were added. After adding β-glucan and pullulan, the stirring speed was 800 r / min, and the stirring time was 20 min, yielding the stress buffer isolating phase.

[0050] S3, Preparation of the collagen signal maintenance phase. The collagen signal maintenance phase consisted of 150.00 g of deionized water, 80.00 g of 1,3-propanediol, 12.00 g of N-acetylhydroxyproline, 0.24 g of hexapeptide-9, and 0.096 g of palmitoyl tripeptide-5. Deionized water and 1,3-propanediol were added to a mixing tank, and the temperature was controlled at 35°C. N-acetylhydroxyproline was added and stirred until dissolved. After confirming that the system temperature did not exceed 35°C, hexapeptide-9 and palmitoyl tripeptide-5 were added, and stirring was continued for 10 min to obtain the collagen signal maintenance phase.

[0051] S4, Sequential Compounding. First, the stress buffer isolation phase is added to the collagen degradation inhibitor compound phase at a rate of 8.00% of the collagen degradation inhibitor compound phase mass per minute, with a stirring speed of 600 r / min during addition. After all the stress buffer isolation phase is added, stirring continues for 20 min. Then, the collagen signal maintenance phase is added at a rate of 3.00% of the mixed system mass per minute, with a stirring speed of 500 r / min during addition, and the system temperature is controlled at 35℃. After all the collagen signal maintenance phase is added, stirring continues for 10 min, and the final system pH is controlled to 5.8, yielding a non-invasive anti-aging active ingredient compound solution. The resulting compound solution is filtered through a 0.45 μm pore size filter membrane and then placed into an opaque container.

[0052] In this embodiment, the mass ratio of hydroxytyrosol to zinc pyrrolidone carboxylate is 1:1.1; the mass ratio of ectoine to ergothioneine is 50:1; the mass ratio of β-glucan to pullulan is 1:0.5; and the mass ratio of N-acetylated hydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 is 100:2.0:0.8.

[0053] Example 4 The non-invasive anti-aging active ingredient compound obtained in Example 1 was added to the serum matrix as an active ingredient additive. The serum matrix includes deionized water, glycerin, butylene glycol, thickener, preservative, and pH adjuster. During preparation, the serum matrix was first prepared and cooled to below 35°C. Then, the non-invasive anti-aging active ingredient compound obtained in Example 1 was added at a rate of 3.00% of the total serum mass. The mixture was stirred until homogeneous, and the final pH was adjusted to 5.4–5.8.

[0054] In this embodiment, the non-invasive anti-aging active ingredient compound solution is added to the skin care matrix as a pre-combined three-phase active ingredient component, instead of directly adding hydroxytyrosol, zinc pyrrolidone carboxylate, ectoine, ergothioneine, N-acetylhydroxyproline, and peptide components to the skin care matrix separately. This addition method avoids interference from thickeners, emulsifiers, or preservatives in the skin care matrix during the three-phase compounding process.

[0055] The non-invasive anti-aging active ingredient compound solution can also be added to emulsion base, cream base, or mask base at a mass percentage of 1.00–6.00%. When adding, the temperature of the skincare base should be controlled at 25–35°C, and the stirring speed should be controlled at 200–500 r / min. Continue stirring for 10–30 minutes after addition. If adding to a cream base, it should be added after emulsification is complete and the temperature has cooled; if adding to a mask base, it should be added after the thickener is fully hydrated and defoamed.

[0056] Comparative Example 1: One-time mixing preparation This comparative example uses the exact same components and amounts as Example 1, but instead of preparing the collagen degradation inhibitory complex phase, stress buffer isolation phase, and collagen signal maintenance phase, deionized water, 1,3-propanediol, hydroxytyrosol, zinc pyrrolidone carboxylate, ectoine, ergothioneine, trehalose, β-glucan, pullulan, N-acetylhydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 were sequentially added to the same mixing tank. The mixture was stirred at 30°C for 60 min, and the final pH was adjusted to 5.5 to obtain the Comparative Example 1 sample. This comparative example is used to illustrate the difference between a one-time mixing method and the three-phase sequential mixing method of this application under the same components and content conditions.

[0057] Comparative Example 2: No pre-combination of hydroxytyrosol and zinc pyrrolidone carboxylate This comparative example uses the same components and amounts as Example 1, but in S1, only hydroxytyrosol is dissolved in an aqueous phase containing 1,3-propanediol, and zinc pyrrolidone carboxylate is not added; zinc pyrrolidone carboxylate is added together with the collagen signal maintenance phase in the final compounding stage in S4, and other conditions are the same as in Example 1. This comparative example is used to illustrate the difference between pre-contacting hydroxytyrosol and zinc pyrrolidone carboxylate in a weakly acidic aqueous phase and adding them separately in the final system.

[0058] Comparative Example 3: Pullulan was removed This comparative example uses the same components and amounts as Example 1, but without pullulan. Deionized water is added to bring the total to 1000g, and other conditions are the same as in Example 1. This comparative example is used to illustrate the difference between the stress buffering and isolating phase formed by β-glucan and pullulan together and the aqueous dispersion system formed by using only β-glucan.

[0059] Comparative Example 4: Peptide components added during the high-temperature stage This comparative example uses the same components and amounts as Example 1, but hexapeptide-9 and palmitoyl tripeptide-5 are added together with hydroxytyrosol and zinc pyrrolidone carboxylate in stage S1, and stirred at 40°C for 45 min. Other conditions are the same as in Example 1. This comparative example is used to illustrate the difference between adding peptide components during the collagen degradation inhibition complex phase pre-preparation stage and adding the collagen signal maintenance phase later at a temperature not exceeding 35°C in this application.

[0060] Comparative Example 5: Zinc pyrrolidone carboxylate was removed This comparative example uses the same components and amounts as Example 1, but without the addition of zinc pyrrolidone carboxylate. Deionized water is added to bring the total to 1000g, and other conditions are the same as in Example 1. This comparative example illustrates the difference between a collagen degradation inhibitory complex containing both hydroxytyrosol and zinc pyrrolidone carboxylate and an aqueous system containing only hydroxytyrosol.

[0061] Comparative Example 6: Removal of N-acetylhydroxyproline This comparative example uses the same components and amounts as Example 1, but without the addition of N-acetylhydroxyproline. Deionized water is added to a final volume of 1000g. Hexapeptide-9 and palmitoyl tripeptide-5 are added as in Example 1, and other conditions are the same as in Example 1. This comparative example illustrates the difference between the collagen signal maintenance phase composed of N-acetylhydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5, and a collagen signal maintenance phase using only peptide components.

[0062] To avoid including unsubstantiated data in the specification, this application only discloses repeatable testing methods and comparative ratio settings. Upon actual submission, if the applicant has already completed the testing, the true test results should be entered into the corresponding form; if the testing has not yet been completed, it is inappropriate to fabricate specific percentage data in the specification.

[0063] Appearance and centrifugal stability testing: Take 50 mL of each sample obtained from Examples 1-3 and Comparative Examples 1-6, place them in transparent centrifuge tubes, and observe for any visible precipitation, flocculation, layering, or uneven coloring. Then centrifuge at 4500 rpm for 30 min, and observe again for any precipitation, layering, or flocculation. This test is used to determine the physical dispersion stability of the system after three-phase sequential compounding. Visual inspection is performed under natural light or a standard light source box, and the sample number, observation temperature, and photographing time are recorded.

[0064] pH measurement: The pH of the example and comparative samples was measured at 25°C using a calibrated pH meter. The pH meter was calibrated with standard buffer solutions of pH 4.00 and pH 6.86 before use. Each sample was measured in triplicate, and the average value was taken. The final pH of the compound solution was controlled within the range of 5.3–5.8.

[0065] Hydroxytyrosol retention rate detection: The hydroxytyrosol content was measured in the example and comparative samples after preparation (0 days) and after being stored at 45°C in the dark for 30 days. The hydroxytyrosol content was determined using high-performance liquid chromatography (HPLC), and the detection conditions were established based on hydroxytyrosol standards. The hydroxytyrosol retention rate was calculated using the following formula: ; in, The percentage of hydroxytyrosol retention is expressed as a percentage (%). This indicates the mass concentration of hydroxytyrosol obtained 0 days after sample preparation; This indicates the mass concentration of hydroxytyrosol obtained after the sample was stored at 45°C in the dark for 30 days. If the sample was diluted during testing, and The mass concentration after conversion using the same dilution factor.

[0066] Color difference detection: Samples from the examples and comparative examples were taken, and the color difference changes were measured after 0 days and 30 days of storage at 45°C in the dark. The color difference was measured using a colorimeter, specifically the L, a, and b values. This test is used to evaluate the color change of the samples before and after high-temperature storage. The test results can be directly recorded as the total color difference value output by the colorimeter, without the need for a separate calculation formula.

[0067] Cell compatibility testing: Human dermal fibroblasts were used for cell compatibility evaluation. The example and comparative samples were added to the cell culture system at predetermined dilutions, and cell viability was detected using the CCK-8 assay or MTT assay after 24 hours. Results were recorded as a percentage of cell viability. This assay was used to confirm cell compatibility at the set dilutions and is not intended to express therapeutic efficacy.

[0068] MMP-1 relative expression detection: The relative expression level of MMP-1 was evaluated using a UV-A-induced human dermal fibroblast model. The model group consisted of cells induced by UV-A but without the addition of a sample; the control group consisted of cells not induced by UV-A and without the addition of a sample; the sample group consisted of cells induced by UV-A and supplemented with samples from the examples or comparative studies. MMP-1 protein content was detected using ELISA, and the results were recorded as the relative expression level of MMP-1 in the sample group compared to the model group.

[0069] COL1A1 relative expression detection: The relative expression level of COL1A1 was evaluated using human dermal fibroblasts. After sample processing, COL1A1 gene expression could be detected by qPCR, with GAPDH or β-actin as the internal control gene. If qPCR was not used, type I collagen ELISA could be used instead, but the same detection method should be used for the same set of examples and comparative examples.

[0070] AGEs relative fluorescence intensity detection: The fluorescence intensity of AGEs was evaluated using a bovine serum albumin-glucose in vitro glycation model. Samples from both examples and comparative studies were added to the glycation reaction system at the same dilution factor. After incubation at a constant temperature, the fluorescence intensity of AGEs was detected using a fluorescence microplate reader. The results were recorded as the relative fluorescence intensity of AGEs in the sample group compared to the model group. This indicator is used to compare the differences between the examples and comparative studies in the in vitro glycation model and is not intended to directly claim therapeutic effects in humans.

[0071] In the above detection methods, hydroxytyrosol retention, color difference, and centrifugal stability are used to verify the effect of the three-phase sequential compounding on the stability of the system; relative expression of MMP-1, relative expression of COL1A1, and relative fluorescence intensity of AGEs are used to verify the compounding logic between the collagen degradation inhibition compounding phase, the stress buffer isolation phase, and the collagen signal maintenance phase. Formal application documents should prioritize the use of actual test results already completed by the applicant and should not fabricate data on the reduction rate of human wrinkles, the increase rate of collagen production, or cell experiment data.

[0072] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A non-invasive anti-aging active ingredient compound solution that improves fine lines and inhibits collagen loss, characterized in that, include: The non-invasive anti-aging active ingredient compound solution is formed by first adding the stress buffer isolation phase to the collagen degradation inhibitory compound phase, and then adding the collagen signal maintenance phase. The collagen degradation inhibitor complex consists of hydroxytyrosol, zinc pyrrolidone carboxylate, 1,3-propanediol, and deionized water; The stress buffering phase includes ectoine, ergothioneine, trehalose, β-glucan, pullulan, and deionized water; The collagen signal maintenance phase includes N-acetylhydroxyproline, hexapeptide-9, palmitoyl tripeptide-5, 1,3-propanediol, and deionized water; Based on the total mass of the non-invasive anti-aging active ingredient compound solution, the non-invasive anti-aging active ingredient compound solution comprises the following components by mass percentage: hydroxytyrosol 0.10–0.60%, zinc pyrrolidone carboxylate 0.07–0.66%, ectoine 1.50–3.00%, ergothioneine 0.03–0.07%, trehalose 2.00–8.00%, β-glucan 0.10–0.40%, pullulan 0.04–0.20%, N-acetylated hydroxyproline 0.60–1.20%, hexapeptide-9 0.006–0.024%, palmitoyl tripeptide-5 0.0024–0.012%, 1,3-propanediol 6.00–18.00%, pH adjuster 0.01–0.30%, with the balance being deionized water.

2. The non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss according to claim 1, characterized in that, The mass ratio of hydroxytyrosol to zinc pyrrolidone carboxylate is 1:0.7 to 1.1; The mass ratio of ectoine to ergothioneine is 30–70:1; The mass ratio of the β-glucan to pullulan is 1:0.2 to 1.

5.

3. The non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss according to claim 2, characterized in that, The mass ratio of N-acetylhydroxyproline, hexapeptide-9, and palmitoyl tripeptide-5 is 100:0.6–2.5:0.2–1.0; The mass ratio of the collagen degradation inhibitory complex phase, the stress buffer isolation phase, and the collagen signal maintenance phase is 1:2.0-5.0:0.3-0.

8.

4. The non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss according to claim 3, characterized in that, The pH adjuster is composed of citric acid and sodium citrate, and the pH of the non-invasive anti-aging active ingredient compound solution is 5.3 to 5.8; The non-invasive anti-aging active ingredient compound solution is filtered through a 0.45 μm pore size filter membrane and then placed into an opaque container.

5. The non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss according to claim 4, characterized in that, The non-invasive anti-aging active ingredient compound solution does not contain retinol active ingredients, hydroxypinazone retinate, hydroxypropyl tetrahydropyranotriol, acetyl hexapeptide-8, or exogenous collagen active ingredients; The retinol-based active ingredients include retinol, retinaldehyde, retinyl palmitate, and retinyl acetate; The exogenous collagen-like active ingredients include hydrolyzed collagen, soluble collagen, and recombinant collagen.

6. A method for preparing a non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss, applicable to the non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss as described in any one of claims 1-5, characterized in that, Includes the following steps: S1: Preparation of collagen degradation inhibitory complex phase: Hydroxytyrosol was added to an aqueous phase containing 1,3-propanediol and stirred to dissolve at 35-45℃. Then, zinc pyrrolidone carboxylate was added, and the pH of the system was adjusted to 5.2-5.6 using a pH adjuster. The mixture was stirred for 30-60 min to obtain the collagen degradation inhibitory complex phase. S2: Preparation of stress buffer isolation phase: Add ectoine, trehalose, ergothioneine, β-glucan and pullulan to the aqueous phase, and stir at 25-35℃ until the system is homogeneous to obtain the stress buffer isolation phase; S3: Preparation of collagen signal maintenance phase: Dissolve N-acetylhydroxyproline in aqueous phase, cool to no higher than 35℃, add hexapeptide-9 and palmitoyl tripeptide-5, stir for 10-25 min to obtain collagen signal maintenance phase; S4: Sequential compounding: First, add the stress buffer isolation phase to the collagen degradation inhibition compound phase and stir for 20-40 minutes. Then, add the collagen signal maintenance phase to the mixed system and control the final system pH to 5.3-5.8 to obtain a non-invasive anti-aging active ingredient compound solution.

7. The preparation method of a non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss according to claim 6, characterized in that, The aqueous phase containing 1,3-propanediol consists of 1,3-propanediol and deionized water, and the mass percentage of 1,3-propanediol in the collagen degradation inhibitory complex phase is 10.00–20.00%. After the hydroxytyrosol is added to the aqueous phase containing 1,3-propanediol, it is stirred at a speed of 300-600 r / min for 10-20 min, and then zinc pyrrolidone carboxylate is added.

8. The preparation method of a non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss according to claim 7, characterized in that, The pH adjuster includes an aqueous solution of citric acid and an aqueous solution of sodium citrate, wherein the concentration of the aqueous solution of citric acid is 0.05–0.20 mol / L and the concentration of the aqueous solution of sodium citrate is 0.05–0.20 mol / L. After adjusting the pH of the system, continue stirring the system at 35–45°C for 30–60 min.

9. The preparation method of a non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss according to claim 8, characterized in that, In step S2, trehalose, ectoine, and ergothioneine are first added to deionized water and stirred to dissolve. Then, β-glucan and pullulan are added and stirred to disperse. The stirring speed after adding the β-glucan and pullulan is 400-800 r / min, and the stirring time is 20-50 min.

10. The preparation method of a non-invasive anti-aging active ingredient compound solution for improving fine lines and inhibiting collagen loss according to claim 9, characterized in that, The mass ratio of the collagen degradation inhibitory complex phase, the stress buffering and isolating phase, and the collagen signal maintenance phase is 1:2.0-5.0:0.3-0.8; In step S4, when the stress buffer isolation phase is added to the collagen degradation inhibitory complex phase, the addition rate is 2.00-8.00% of the mass of the collagen degradation inhibitory complex phase per minute, and the stirring speed during the addition is 300-600 r / min. When the collagen signal maintenance phase is added to the mixing system, the temperature of the mixing system is 25-35℃, the addition rate is 0.50-3.00% of the mass of the mixing system per minute, and the stirring speed during the addition is 200-500 r / min; After all the collagen signal maintenance phase is added, continue stirring for 10–30 minutes.

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