Anti-oxidation glowing mask technology combining conopeptide and ergothioneine

Abstract

The invention relates to the technical field of masks, and discloses a conopeptide and ergothioneine combined antioxidant glowing mask process which comprises the following components in percentage by mass: an active component composition: 0.01-0.1% of liposome coated conopeptide and 0.2-1.0% of liposome coated ergothioneine; the matrix components comprise 50-70% of water, 3-8% of glycerol, 2-5% of butanediol, 2-5% of nicotinamide, 1-5% of squalane and 3-8% of meadowfoam seed oil; the emulsifying agent is prepared from the following components in percentage by weight: 0.5 to 2 percent of polyglycerol-10 laurate, 0.5 to 1.5 percent of sorbitol polyether-30 tetraoleate and 0.1 to 0.5 percent of sodium polyacryloyldimethyl taurate. According to the invention, the direct contact between the active peptide and the antioxidant is blocked through the combination of double-cabin isolated packaging and a liposome freeze-drying technology, and chain degradation caused by mutual exclusion of components is thoroughly avoided through physical separation and glassy state curing.

Description

Technical Field

[0001] The present invention relates to the technical field of facial masks, and specifically to an antioxidant and radiance-enhancing facial mask process combining conotoxin peptide and ergothioneine. Background Art

[0002] In the field of antioxidant and radiance-enhancing facial masks, bioactive peptides (conotoxin peptide) and natural antioxidants (ergothioneine) are widely used as core active ingredients. The existing technology mainly relies on a liquid single-chamber formulation system, disperses the active substances through a conventional emulsification process, and uses low-temperature freeze-drying or hot-melt filling for final product shaping. The industry standard process covers basic production processes such as the compounding of polyol humectants, the stability control of a single pH regulator (lactic acid), and open homogenization filling.

[0003] In the existing technology, the storage in a liquid single chamber causes the hydrolysis of the β-sheet conformation of conotoxin peptide (the amide bond breakage rate > 30%), and ergothioneine generates quinone derivatives due to the penetration of dissolved oxygen (the ORAC value decays by 67%). In the traditional freeze-drying process, the temperature in the sublimation stage > 30°C, the liposome membrane rupture rate exceeds 20%, and the transdermal delivery efficiency is less than 1 / 3 of the standard requirement. The single pH regulator lacks the ability of metal chelation, and Fe 3+ catalyzes the free radical chain reaction, resulting in the oxidation inactivation of the formulation (the activity retention rate < 60%). The residual oxygen concentration in the atmospheric pressure filling process > 2 ppm, the peroxide value (PV) of squalane soars to 5 meq / kg within 6 months, and at the same time, the growth of microorganisms (CFU > 10 3 ) forces the excessive addition of preservatives, increasing the irritation risk. Summary of the Invention

[0004] Aiming at the deficiencies of the existing technology, the present invention provides an antioxidant and radiance-enhancing facial mask process combining conotoxin peptide and ergothioneine, which solves the problems of mutual exclusion and degradation of active substances in the liquid single-chamber formulation in the existing technology, the damage of the integrity of the liposome membrane by the traditional freeze-drying process, the inability of a single acid regulator to inhibit metal ion-catalyzed oxidation, and the rancidity of oils and fats and the growth of microorganisms caused by residual oxygen in atmospheric pressure filling.

[0005] To achieve the above objectives, the present invention is realized through the following technical solutions: an antioxidant and radiance-enhancing facial mask process combining conotoxin peptide and ergothioneine, including the following components in mass fraction:

[0006] Active ingredient composition: liposome-encapsulated conotoxin peptide 0.01 - 0.1%, liposome-encapsulated ergothioneine 0.2 - 1.0%;

[0007] Liposome-encapsulated conotoxin peptide (0.01 - 0.1%):

[0008] Conus peptides inhibit the release of acetylcholine by blocking the muscle sodium ion channel (Nav1.4), reducing the transmission of nerve signals to facial muscles, thereby instantly relaxing the mimetic muscles and alleviating dynamic wrinkles; long-term use can stimulate fibroblasts to secrete type I collagen and elastic fibers, enhance the supporting structure of the dermis, and improve skin firmness. Liposome encapsulation (phospholipid bilayer) can enhance its transdermal absorption efficiency, prevent peptide chains from being degraded by proteases, and achieve a sustained release effect.

[0009] Liposome-encapsulated ergothioneine (0.2 - 1.0%):

[0010] As an endogenous antioxidant, ergothioneine enters cells through the specific transporter OCTN1, directly scavenges hydroxyl radicals (·OH) and peroxynitrite (ONOO-), and simultaneously activates the Nrf2 pathway, upregulating the synthesis of thioredoxin (Trx) and glutathione (GSH), and repairing UV-induced DNA damage. Liposome encapsulation can improve its photostability and enhance the targeted delivery efficiency through a bilayer structure similar to skin lipids.

[0011] Matrix components: water 50 - 70%, glycerin 3 - 8%, butylene glycol 2 - 5%, niacinamide 2 - 5%, squalane 1 - 5%, Limnanthes alba seed oil 3 - 8%;

[0012] Components and ratios:

[0013] Water (50 - 70%), glycerin (3 - 8%), butylene glycol (2 - 5%):

[0014] Glycerin and butylene glycol form a polyol moisturizing network, bind water molecules through hydrogen bonds, reduce the water activity of the system (Aw < 0.6), and inhibit the proliferation of microorganisms; at the same time, butylene glycol as a penetration enhancer can temporarily disrupt the lipid arrangement of the stratum corneum and enhance the penetration of active ingredients.

[0015] Niacinamide (2 - 5%):

[0016] Niacinamide (vitamin B3), as a NAD+ / NADH precursor, reduces pigment deposition by inhibiting melanosome transport, promotes ceramide synthesis, and repairs the skin barrier; its anti-glycation effect can inhibit the cross-linking and destruction of collagen by advanced glycation end products (AGEs).

[0017] Squalane (1 - 5%), Limnanthes alba seed oil (3 - 8%):

[0018] Squalane highly mimics the components of the sebum membrane, fills the gaps in the stratum corneum, and enhances the barrier function; Limnanthes alba seed oil contains long-chain fatty acids (C20 - 24), forms a dense oil film, reduces transepidermal water loss (TEWL), and regulates the activity of sebaceous glands through ω-6 linoleic acid to relieve inflammation.

[0019] Emulsifier: Polyglyceryl-10 laurate 0.5 - 2%, sorbitan polyether-30 tetraoleate 0.5 - 1.5%, sodium polyacryloyldimethyltaurate 0.1 - 0.5%;

[0020] Components and ratios:

[0021] Polyglyceryl-10 laurate is a non-ionic emulsifier. Its polyglycerol chain can form a lamellar liquid crystal structure to wrap the oil-phase droplets, enhancing the low-temperature stability of the emulsion (no demulsification at -20°C); sorbitan polyether-30 tetraoleate can achieve the preparation of high internal phase emulsion (HIPE) by reducing the interfacial tension (γ < 1 mN / m), increasing the oil-phase loading capacity; sodium polyacryloyldimethyltaurate is an anionic polymer, which can prevent emulsion flocculation through electrostatic repulsion and form a thixotropic gel network to enhance the spreadability of the product.

[0022] Stabilizer: 1,2-hexanediol 0.5 - 2%, p-hydroxyacetophenone 0.1 - 0.5%, octanoyl hydroxamic acid 0.05 - 0.2%;

[0023] Components and ratios:

[0024] 1,2-Hexanediol achieves broad-spectrum antibacterial by disrupting the phospholipid bilayer of microbial cell membranes (LogP = 1.2). At the same time, p-hydroxyacetophenone, as a radical terminator, preferentially combines with alkoxy radicals (RO·) in the lipid peroxidation chain reaction to block the oxidation reaction process; octanoyl hydroxamic acid inhibits the generation of hydroxyl radicals in the Fenton reaction by chelating metal ions (Fe 2+ , Cu 2+ ), and synergizes with 1,2-hexanediol to pass the EU anti-corrosion challenge test (ISO29621 standard).

[0025] Preferably, the active ingredient composition further includes at least one of the following components:

[0026] Palmitoyl tripeptide-1: 0.01 - 0.1%;

[0027] Superoxide dismutase: 500 - 5000 U / g;

[0028] Fullerene: 0.05 - 0.3%.

[0029] 1. Palmitoyl tripeptide-1 (0.01 - 0.1%):

[0030] Palmitoyl tripeptide-1 activates the transforming growth factor-β (TGF-β) signaling pathway, upregulates the synthesis of type I collagen, type III collagen, and fibronectin in dermal fibroblasts, and simultaneously inhibits the activity of matrix metalloproteinase (MMP-1) to reduce collagen degradation. Its palmitoylation modification enhances liposolubility, promotes penetration through the stratum corneum, targets the dermis layer, and forms a complement to the sodium ion channel blocking mechanism of conotoxin peptides. It achieves long-term anti-wrinkle effects through a dual pathway (signal transduction inhibition + collagen synthesis promotion).

[0031] 2. Superoxide dismutase (SOD, 500 - 5000 U / g):

[0032] SOD catalyzes the disproportionation of superoxide anion radicals (O2 - ·) into hydrogen peroxide (H2O2) and oxygen (O2), directly scavenging the primary free radicals induced by ultraviolet rays and environmental pollution; it forms an "oxidative chain reaction interception system" with ergothioneine - ergothioneine repairs DNA damage, SOD blocks the source of free radicals, and the two work together to reduce the cellular oxidative stress level (ROS) below the threshold (<100 nM), preventing the activation of the NF -k B inflammatory pathway and reducing collagen loss.

[0033] 3. Fullerene (C60, 0.05 - 0.3%):

[0034] Fullerene efficiently adsorbs a variety of free radicals (including ·OH, O2 - ·, ROO·) through its cage - like cavity structure. A single molecule can continuously capture 34 free radicals (sponge - like effect), and its antioxidant capacity is 172 times that of vitamin C; its liposoluble property (LogP = 6.5) enables it to accumulate in the sebum membrane, forming a long - acting free radical scavenging reserve pool (action time > 12 hours), and forms a cross - phase antioxidant matrix with ergothioneine (water - soluble) to cover the hydrophilic / lipophilic regions of the skin.

[0035] Preferably, the matrix component further includes at least one of the following components:

[0036] Trehalose: 1 - 3%;

[0037] Jojoba seed oil: 1 - 3%;

[0038] Grape seed oil: 0.5 - 2%;

[0039] Sodium hyaluronate: 0.1 - 0.5%.

[0040] 1. Trehalose (1 - 3%):

[0041] Trehalose forms a "water substitution" effect through its unique disaccharide cyclic structure. When the skin is dehydrated, it binds to phospholipid membranes and proteins via hydrogen bonds, preventing the phase transition of the lipid bilayer and protein thermal denaturation, and maintaining the structural integrity of keratinocytes. At the same time, it activates the autophagy pathway (AMPK / mTOR), clears damaged mitochondria, and enhances the survival rate of epidermal cells under ultraviolet radiation.

[0042] 2. Jojoba seed oil (1 - 3%):

[0043] Jojoba seed oil contains more than 97% long-chain wax esters (C36 - C46), and its molecular structure is highly similar to human sebum (squalene, wax esters). It repairs the barrier by filling the lipid cavities in the stratum corneum, reducing the transepidermal water loss rate (TEWL) by ≥25%. The natural tocopherols (≥0.1%) and polyphenolic substances contained in it quench singlet oxygen ( 1 O2), inhibit the abnormal follicular keratinization caused by sebum oxidation, and reduce the formation of closed comedones.

[0044] 3. Grape seed oil (0.5 - 2%):

[0045] Proanthocyanidins (OPC ≥ 95%) in grape seed oil inhibit the activity of matrix metalloproteinase-2 (MMP-2), blocking the degradation of type IV collagen induced by ultraviolet rays. Its high content of linoleic acid (ω-6, content ≥ 70%) acts as a precursor of prostaglandin E2 (PGE2), regulating the PPARγ receptor of sebaceous gland cells, normalizing the ratio of squalene / triglyceride in sebum composition, and improving the microenvironment of oily skin.

[0046] 4. Sodium hyaluronate (0.1 - 0.5%):

[0047] Sodium hyaluronate forms a breathable moisturizing film on the epidermis through a molecular weight gradient design (high molecular weight (1 - 1.8 MDa)), and the low molecular weight (50 - 300 kDa) penetrates into the dermis to activate the CD44 receptor, stimulating the migration and proliferation of fibroblasts. It forms a three-dimensional hydration network with trehalose, increasing the water content of the stratum corneum by ≥40% (measured by Corneometer), and alleviating the skin dryness side effect after injection of botulinum toxin type A.

[0048] Preferably, the liposome-encapsulated conotoxin and ergothioneine liposome comprises:

[0049] Hydrogenated lecithin: 2 - 5%;

[0050] Cholesterol: 0.5 - 1.5%;

[0051] Mannitol: 1 - 3%.

[0052] 1. Hydrogenated lecithin (2 - 5%):

[0053] Hydrogenated lecithin forms a high-density lipid bilayer membrane through its saturated fatty acid chains (mainly C16:0), raising the phase transition temperature (Tm) to 55 - 65 °C, significantly enhancing the membrane structure stability of liposomes during high-temperature storage (40 °C), and preventing the leakage of active ingredients (leakage rate < 5% / month); its amphiphilic structure (HLB = 8 - 10) can reduce the oil-water interfacial tension to < 3 mN / m, promoting the fusion of liposomes with stratum corneum lipids and increasing the transdermal absorption efficiency by 2.3 times (measured data from Franz diffusion cells).

[0054] 2. Cholesterol (0.5 - 1.5%):

[0055] Cholesterol inserts into the phospholipid bilayer at a molar ratio of 1:2. By restricting the lateral movement of phosphatidylcholine molecular chains through its rigid steroid ring structure, it increases the membrane microviscosity (η) of liposomes from 0.89 Pa·s to 1.35 Pa·s, inhibiting liposome aggregation (particle size change rate < 10% @ 25 °C / 6 months); at the same time, its hydroxyl group forms hydrogen bonds with stratum corneum ceramides, guiding liposomes to target weak areas of the skin barrier (wrinkle grooves), and improving the delivery accuracy by 40%.

[0056] 3. Mannitol (1 - 3%):

[0057] Mannitol forms a glassy matrix during freeze-drying (glass transition temperature Tg = -30 °C), fixing the spatial conformation of liposomes through a hydrogen bond network and preventing ice crystal growth from damaging the membrane structure (freeze-drying survival rate > 95%); its hexitol structure preferentially binds to water molecules during reconstitution (hydration energy -25.1 kJ / mol), accelerating the dispersion of liposomes to the nanoscale (particle size growth < 15 nm after reconstitution), and ensuring the instantaneous release of active substances (T90% release time ≤ 5 min).

[0058] Preferably, the liposomes have a particle size of 50 - 100 nm and are prepared by a high-pressure microfluidization homogenization process with a homogenization pressure of 15,000 - 20,000 psi and a circulation number of 3 - 5 times.

[0059] 1. Liposome particle size (50 - 100 nm):

[0060] Liposomes with a particle size of 50 - 100 nm can precisely match the intercellular spaces of skin stratum corneum cells (about 30 - 50 nm) and the size of hair follicle openings (about 100 - 300 μm), and penetrate through both passive diffusion (Stokes-Einstein equation) and hair follicle bypass pathways, with the transdermal efficiency being 3.2 times higher than that of micron-sized liposomes (verified by in vitro transdermal experiments); at the same time, this particle size range enables the specific surface area (SSA) of liposomes to reach 60 - 80 m 2 / g, ensuring an active ingredient encapsulation rate > 90% and an aggregation rate < 5% during the storage period (dynamic light scattering monitoring data).

[0061] 2. High-pressure microfluidization homogenization process (homogenization pressure 15,000 - 20,000 psi):

[0062] Under the ultra-high pressure of 15,000 - 20,000 psi, the liposome premix generates an instantaneous shear rate (>10 7 s -1 ) and cavitation effect through the counter-impact of microfluidics, breaking the liposome particles into nanoscale (Weber number We > 10 4 ), and at the same time, the high-pressure impact reorients the polar head groups of phospholipid molecules to form a monolayer dispersion system (PDI < 0.2), avoiding the temperature sensitivity defect of traditional homogenizers (process temperature < 35°C, active ingredient degradation rate < 1%).

[0063] 3. Number of cycles (3 - 5 times):

[0064] 3 - 5 cycles eliminate the bimodal particle size distribution (the span value of the particle size distribution drops from 1.5 to 0.8) through the repeated process of fragmentation - recombination, making the coefficient of variation (CV) of the liposome particle size < 15%; more than 5 cycles will cause excessive stretching of phospholipid molecules (membrane tension > 30 mN / m), leading to perforation of the liposome membrane (leakage rate > 8%), while less than 3 cycles cannot fully disperse the aggregates (the proportion of particles with a residual particle size > 200 nm > 20%).

[0065] Preferably, the preparation process of the facial mask includes the following steps:

[0066] Heat the aqueous phase components to 40 - 50°C for dissolution, and then add the active ingredient composition after cooling to 25 - 30°C;

[0067] Heat the oil phase components (squalane, Limnanthes alba seed oil, Simmondsia chinensis seed oil, Vitis vinifera seed oil) to 40 - 50°C and mix evenly;

[0068] Mix the aqueous phase and the oil phase in a vacuum homogenizer, with a homogenization pressure of 10 - 15 MPa and a mixing time of 15 - 30 min;

[0069] Inert gas protection is adopted during the final product mixing and filling process.

[0070] 1. Treatment of aqueous phase components (heat to 40 - 50°C for dissolution, add active ingredients after cooling to 25 - 30°C):

[0071] The aqueous components (water, glycerin, butanediol) are completely dissolved at 40 - 50 °C. This temperature range is higher than the melting point of glycerin (17.8 °C) but lower than the thermal decomposition threshold of niacinamide (>60 °C), ensuring dissolution efficiency while avoiding degradation of heat-sensitive components. After cooling to 25 - 30 °C, the active ingredients (SOD, fullerene) are added, which can avoid enzyme inactivation caused by high temperature (the active half-life of SOD is shortened by 50% at >35 °C) and the formation of fullerene aggregates (the surface tension decreases and the dispersion degree increases at low temperature).

[0072] 2. Treatment of oil-phase components (mixing squalane and Limnanthes alba seed oil by heating to 40 - 50 °C):

[0073] Mechanism: Squalane (freezing point -12 °C) and Limnanthes alba seed oil (freezing point 10 °C) reach a kinematic viscosity < 35 mm 2 / s at 40 - 50 °C, forming Newtonian fluid characteristics to ensure the uniformity of the oil phase; the oxidation rate of polyunsaturated fatty acids (linoleic acid) in jojoba seed oil and grape seed oil is < 0.01% / h (tested by the Rancimat method) at this temperature, avoiding thermal-induced rancidity.

[0074] 3. Vacuum homogenization mixing (pressure 10 - 15 MPa, time 15 - 30 min):

[0075] Mechanism: The vacuum environment (-0.08 to -0.1 MPa) removes dissolved oxygen (residual oxygen concentration < 0.5 ppm), inhibiting the free radical chain reaction; the homogenization pressure of 10 - 15 MPa reduces the droplet size D50 of the oil phase to 200 - 500 nm (laser particle size analysis), forming an O / W emulsion with an interfacial film thickness ≥ 10 nm (measured by SAXS). Homogenization at low temperature (30 - 40 °C) avoids the HLB value drift of the emulsifier (polyglyceryl-10 laurate) (ΔHLB < 0.5).

[0076] 4. Inert gas protection filling (nitrogen covering):

[0077] Mechanism: Nitrogen (purity ≥ 99.999%) is continuously introduced during the filling process to control the headspace oxygen concentration < 0.1% (monitored by an oxygen sensor), blocking the oxidation of ergothioneine (the oxidation half-life is extended to > 18 months), and maintaining the peroxide value (PV) of squalane < 1 meq / kg (detected by the GB5009.227 method); at the same time, the nitrogen flow rate of 0.5 - 1.0 L / min forms a laminar flow covering to prevent bubble entrainment during filling (the bubble volume ratio < 0.01%).

[0078] Preferably, the vacuum degree of the vacuum homogenizer is -0.08 to -0.1 MPa, and the stirring speed is 500 - 1000 rpm.

[0079] 1. Vacuum degree (-0.08 to -0.1 MPa):

[0080] The vacuum degree of -0.08 to -0.1 MPa (corresponding to an absolute pressure of 20 - 40 kPa) reduces the dissolved oxygen concentration to <0.5 ppm through Henry's Law, inhibiting the oxidative degradation of the active ingredient (ergothioneine) (the oxidation rate is reduced to 1 / 4 under normal pressure); meanwhile, the negative pressure environment causes the microbubbles (diameter <50 μm) in the emulsion to rapidly expand and rupture under the action of Laplace pressure (ΔP = 2γ / r), avoiding the looseness of the paste structure caused by the retention of bubbles.

[0081] 2. Stirring speed (500 - 1000 rpm):

[0082] The shear rate of 500 - 1000 rpm (γ = 10 3 -10 4 s -1 ) regulates the rheology of the emulsion through the Stormer equation (η = Kγ n-1 ), making the oil - phase droplets break to D90 < 5 μm (verified by dynamic light scattering), and at the same time avoiding the damage to the thermosensitive ingredient (liposome) caused by the local temperature rise (ΔT < 2℃) induced by too high a rotation speed (>1200 rpm); this speed range matches the pumping efficiency of the vacuum pump (20 - 40 L / min), ensuring the dynamic balance of the gas - liquid two - phase and preventing the paste from entrapping air due to eddy currents.

[0083] Preferably, the mask adopts a double - chamber packaging, including:

[0084] Compartment A: The liquid component containing the aqueous matrix and the emulsifier;

[0085] Compartment B: The freeze - dried powder containing conotoxin and ergothioneine encapsulated by liposomes.

[0086] Compartment A (aqueous matrix and emulsifier liquid component):

[0087] The aqueous matrix (water, glycerol, butanediol) and the emulsifier (polyglyceryl - 10 laurate, sorbitan polyether - 30 tetraoleate) in Compartment A pre - form a homogeneous liquid phase, and its pH value (5.5 - 6.0) and ionic strength (conductivity < 100 μS / cm) are precisely regulated to avoid the rupture of the liposome membrane caused by osmotic pressure difference during long - term storage (the leakage rate before re - dissolving the freeze - dried powder < 0.5%). The emulsifier forms micelles through pre - hydration (critical micelle concentration CMC = 0.01 - 0.05%), and rapidly disperses the liposomes in the freeze - dried powder during re - dissolution, achieving nano - level redispersion.

[0088] Compartment B (liposome - encapsulated conotoxin and ergothioneine freeze - dried powder):

[0089] Mechanism: The lyophilized powder locks the active ingredients encapsulated in liposomes in a glassy matrix through low-temperature (-40°C pre-freezing) and low-pressure (≤10 Pa sublimation) processes. The water activity (Aw) is reduced to below 0.1, completely blocking the hydrolysis reaction (the hydrolysis rate of conotoxin peptide bonds is reduced to 1 / 20 of that in liquid storage); the lyoprotectant mannitol fixes the spatial conformation of liposomes through a hydrogen bond network, rapidly absorbs free water in the aqueous phase during reconstitution (hydration energy -25.1 kJ / mol), completes the rehydration of liposomes within 5 seconds, and immediately releases the active ingredients.

[0090] Dual-chamber synergistic effect:

[0091] Stability of active substances: Physical isolation of the A / B chambers blocks the potential amidation reaction between conotoxin and nicotinamide (in chamber A) (avoiding the formation of brownish-yellow by-products under high temperature and high humidity);

[0092] Maximization of efficacy: Immediately upon reconstitution, liposomes are oriented at the oil-water interface through interfacial adsorption of emulsifier micelles (contact angle θ = 85 - 90°), and the transdermal efficiency is increased by 1.8 times compared to the premixed emulsion (comparison of the cumulative permeation amount in 24 hours in a Franz diffusion cell);

[0093] Antiseptic exemption: Dual-chamber packaging makes both chamber A (water activity Aw = 0.96) and chamber B (Aw = 0.1) lower than the microbial growth threshold (bacterial Aw > 0.91, mold Aw > 0.80), and it can pass the ISO29621 antiseptic challenge test without adding traditional antiseptics.

[0094] Preferably, the preparation conditions of the lyophilized powder are:

[0095] Pre-freezing temperature: -40°C to -30°C, pre-freezing time: 2 - 4 hours;

[0096] Sublimation temperature: 20 - 25°C, sublimation time: 8 - 12 hours, vacuum degree ≤ 10 Pa;

[0097] Reconstitution temperature of the lyophilized powder: 25 - 30°C, stirring speed: 200 - 400 rpm.

[0098] 1. Pre-freezing conditions (-40°C to -30°C, 2 - 4 hours):

[0099] Mechanism: The pre-freezing temperature of -40°C to -30°C enables the aqueous phase to quickly pass through the maximum ice crystal formation zone (-1°C to -38°C), controlling the ice crystal size to < 20 μm (observed by scanning electron microscopy), avoiding puncture of the liposome membrane by large ice crystals (membrane breakage rate < 3%); 2 - 4 hours of pre-freezing ensures complete vitrification of the system (glass transition temperature Tg = -35°C), and the residual unfrozen water content < 0.5% (determined by the Karl Fischer method), preventing osmotic rupture of liposomes caused by local solute concentration during freeze-drying.

[0100] 2. Sublimation conditions (20 - 25°C, 8 - 12 hours, vacuum ≤ 10 Pa):

[0101] During the sublimation stage, control the temperature at 20 - 25°C (lower than the liposome membrane phase transition temperature of 55°C). By adjusting the temperature of the heating plate, maintain the ice sublimation interface temperature (Tice) at -25°C to -20°C to avoid the phase transition of the liposome bilayer membrane; a vacuum ≤ 10 Pa reduces the sublimation point of ice from 0°C to -50°C (Clapeyron - Clausius equation), accelerating the water removal rate to 1.2 - 1.5 kg / m 2 ·h (monitored by the freeze - drying curve) to ensure that the water content of the lyophilized powder is ≤ 1.5%.

[0102] 3. Reconstitution conditions (25 - 30°C, stirring speed 200 - 400 rpm):

[0103] The reconstitution temperature of 25 - 30°C is close to the lower limit of the liposome membrane phase transition temperature. Through thermal perturbation, promote the movement of the phospholipid molecule tails (membrane fluidity is increased to 0.35 - 0.45 cP), accelerating the penetration of water molecules into the pores of the lyophilized powder (reconstitution time ≤ 30 s); a stirring speed of 200 - 400 rpm corresponds to a Reynolds number Re = 500 - 1000 (from laminar flow to transitional flow), providing an appropriate shear force (τ = 0.1 - 0.5 Pa) to evenly disperse the liposomes and avoid nanoparticle collision and aggregation caused by high - speed stirring (> 500 rpm) (PDI ≤ 0.25 after reconstitution).

[0104] Preferably, the pH value of the facial mask is 5.5 - 6.0 and is adjusted by lactic acid or citric acid;

[0105] 1. pH value range (5.5 - 6.0):

[0106] Mechanism: pH 5.5 - 6.0 is close to but slightly higher than the natural skin pH value (4.5 - 5.5). By creating a weakly acidic environment, reduce the interference with the barrier function (TEWL fluctuation < 5%), while avoiding irritation caused by excessive acidity (EI < 1.5). This range ensures the stability of the liposome membrane (the isoelectric point of phosphatidylcholine is pH = 6.3), prevents aggregation caused by the inversion of the membrane surface charge (Zeta potential > - 30 mV), and maintains the integrity of the α - helix structure of conotoxin (isoelectric point pH = 4.8 - 5.2).

[0107] 2. Lactic acid / citric acid regulation system:

[0108] Lactic acid (pKa = 3.86): As an α-hydroxy acid (AHA), it provides the ability to slowly release hydrogen ions through weak dissociation (dissociation degree < 5% at pH 5.5), balancing the pH fluctuation of the dynamic equilibrium system (ΔpH < 0.2 / 6 months); its molecular structure is similar to the natural moisturizing factor (NMF) of the skin, promoting the degradation of filaggrin in the stratum corneum to generate free amino acids and enhancing the moisturizing performance.

[0109] Citric acid (pKa1 = 3.13): Chelates metal ions (Fe 3+ , Al 3+ ) through its tricarboxylic acid structure, inhibiting the hydroxyl radicals generated by the Fenton reaction (Fe 2+ + H2O2 → ·OH), increasing the oxidation stability of the system by 2.3 times (ORAC value ≥ 8000 μmol TE / g); at the same time, its weak acidity activates the TRPV1 ion channel of keratinocytes, promoting the penetration of active ingredients.

[0110] The present invention provides an antioxidant and radiance-enhancing mask process combining conotoxin and ergothioneine. It has the following beneficial effects:

[0111] 1. The present invention blocks the direct contact between active peptides and antioxidants through double-chamber isolation packaging combined with liposome freeze-drying technology, reducing the inactivation rate of ingredients during the shelf life to less than 5%. In the existing liquid single-chamber formula, more than 25% of the efficacy is lost due to amidation hydrolysis. The present invention completely avoids the chain degradation caused by ingredient incompatibility through physical separation and vitrification.

[0112] 2. The present invention increases the transdermal absorption rate to 3.2 times that of the traditional microemulsion system through nano-liposome particle size control and high-pressure microfluidization homogenization. The traditional technology is limited by insufficient dermal retention of liposomes above 200 nm (< 5%). The present invention realizes 72-hour slow release and targeted deposition in the stratum corneum through shear rate optimization and phase transition temperature regulation.

[0113] 3. The present invention establishes a dynamic equilibrium of pH 5.5 - 6.0 through a lactic acid-citric acid composite buffer system, with a metal ion chelation rate exceeding 95%. The industry pain point of membrane rupture caused by strong acid / weak base environments is solved by the double-carboxylic acid synergistic mechanism. The retention rate of active substances during high-temperature storage increases from 60% to 98%, and the irritation is reduced by 40%.

[0114] 4. The present invention keeps the peroxide value (PV) of grease suppressed below 0.5 meq / kg for a long time through vacuum gradient homogenization combined with nitrogen instantaneous filling. The problem of excessive rancidity caused by residual dissolved oxygen (> 2 ppm) in open production is ended by the vacuum deoxidation (oxygen concentration < 0.1 ppm) and inert gas laminar flow protection of the present invention, and there is no risk of oxidation and deterioration for a long time. Description of the Drawings

[0115] Figure 1Schematic diagram of the method flow of the present invention. Detailed implementation manners

[0116] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without making creative efforts belong to the scope of protection of the present invention.

[0117] Example 1 (anti-wrinkle repair type):

[0118] Formulation (total proportion of A cabin / B cabin is 100%):

[0119] A cabin (liquid): Deionized water 82.5%, glycerin 5%, butanediol 3%, polyglyceryl-10 laurate 1.5%, lactic acid 0.3%, citric acid 0.2%, phenoxyethanol 0.5%;

[0120] B cabin (powder): Liposome-encapsulated conotoxin 2% (containing hydrogenated lecithin 3%, cholesterol 1%, mannitol 2%), freeze-dried ergothioneine 1.5%, mannitol 4.5%.

[0121] Process:

[0122] A cabin: The aqueous phase is dissolved at 45°C and cooled to 28°C, and lactic acid / citric acid is added;

[0123] B cabin: The liposome premix is pre-frozen at -35°C for 3 hours and sublimated at 20 Pa for 10 hours;

[0124] When dispensing, the A / B cabins are covered with nitrogen (oxygen concentration < 0.1%);

[0125] Redissolve and stir at 300 rpm for 30 seconds, and the particle size is restored to 55 nm (PDI = 0.18).

[0126] Example 2 (whitening and antioxidant type):

[0127] Formulation:

[0128] A cabin (liquid): Water 78%, niacinamide 5%, glycerin 4%, sorbitan polyether-30 tetraoleate 2%, citric acid 0.4%;

[0129] B cabin (powder): Liposome-encapsulated ergothioneine 3% (particle size 65 nm), ascorbyl glucoside 2%, trehalose 5%, sodium hyaluronate 0.6%.

[0130] Process:

[0131] The aqueous phase of the A cabin is mixed at 50°C, and the pH is adjusted to 5.8;

[0132] Freeze-drying in Chamber B: Pre-freeze at -38°C for 2.5 hours and sublime at 15 Pa for 12 hours;

[0133] Vacuum homogenization pressure: 12 MPa for 20 minutes, oil droplet D50 = 280 nm;

[0134] The transdermal amount of ergothioneine after reconstitution reaches 0.32 μg / cm 2 / h (0.11 μg for traditional emulsion).

[0135] Example 3 (sensitive skin soothing type):

[0136] Formula:

[0137] Chamber A (liquid): 85% water, 0.5% bisabolol, 2% panthenol, 3% glycerol, 1.5% polyglyceryl-6 laurate;

[0138] Chamber B (powder): 1% liposome-encapsulated ceramide (containing 0.6% cholesterol), 3% centella asiatica extract, 3.4% mannitol.

[0139] Process:

[0140] The emulsifier in Chamber A is pre-hydrated to form micelles (particle size 15 nm);

[0141] The temperature is controlled at 22°C during the sublimation stage of freeze-drying in Chamber B (membrane phase transition protection);

[0142] Stir at 250 rpm during reconstitution, and the encapsulation efficiency of ceramide is 96.7%;

[0143] Example 4 (anti-acne and oil-control type):

[0144] Formula:

[0145] Chamber A (liquid): 76% water, 1.5% salicylic acid, 1% zinc gluconate, 5% glycerol, 2% PEG-40 hydrogenated castor oil;

[0146] Chamber B (powder): 2% liposome-encapsulated tea tree oil (containing 4% lecithin), 3% nano-zinc oxide, 9.5% tapioca starch.

[0147] Process:

[0148] The pH of Chamber A is 5.6 (adjusted with citric acid) to avoid salicylic acid crystallization;

[0149] The water content of the freeze-dried powder in Chamber B is ≤1.2% (inhibiting microbial activity);

[0150] The tea tree oil is slowly released for 12 hours after reconstitution (zero irritating volatile loss).

[0151] Example 5 (anti-blue light repair type):

[0152] Formula:

[0153] Compartment A (liquid): water 80%, ergothioneine 0.8%, ectoin 1%, glycerol 5%, xanthan gum 0.2%;

[0154] Compartment B (powder): liposome-encapsulated lutein 2% (containing cholesterol 1.2%), astaxanthin 1%, microcrystalline cellulose 9%, β-glucan 1%.

[0155] Process:

[0156] Compartment A is vacuum-deoxygenated until the dissolved oxygen < 0.3 ppm;

[0157] Compartment B is freeze-dried: pre-frozen at -40°C for 4 hours and sublimated at 10 Pa for 10 hours.

[0158] Comparative Example 1 (single-compartment mixed type):

[0159] Corresponding to Example 1: The double-compartment design is cancelled, and the components of Compartments A / B are directly mixed.

[0160] Preparation process:

[0161] The liquid components of Compartment A in Example 1 (deionized water 82.5%, glycerol 5%, butanediol 3%) are directly mixed with the freeze-dried powder of Compartment B (liposome-encapsulated conotoxin 2%, ergothioneine 1.5%) at 25°C;

[0162] The homogenization pressure after mixing is 10 MPa / 20 minutes, and nitrogen protection is not carried out;

[0163] After filling, it is stored under accelerated conditions of 40°C / 75% RH.

[0164] Comparison point: The active ingredients are directly in contact with the liquid environment and are not stored separately to verify the necessity of the double-compartment to block chemical reactions.

[0165] Comparative Example 2 (traditional freeze-drying process):

[0166] Corresponding to Example 2: The conventional freeze-drying process is adopted (sublimation temperature > 30°C).

[0167] Preparation process:

[0168] When preparing the freeze-dried powder of Compartment B, the temperature of the heating plate is raised to 32°C during the sublimation stage, and the vacuum degree is maintained ≤ 10 Pa;

[0169] The remaining steps are the same as those in Example 2 (including pH adjustment and vacuum homogenization of Compartment A);

[0170] The reconstitution stirring speed is 500 rpm.

[0171] Comparison point: High-temperature sublimation destroys the liposome membrane structure to verify the protective effect of the sublimation temperature (20 - 25°C) of the present invention on the membrane integrity.

[0172] Comparative Example 3 (pH adjustment with a single acid):

[0173] Corresponding to Example 3: Only lactic acid was used to adjust the pH to 5.5, and citric acid was cancelled.

[0174] Preparation process:

[0175] Only 0.5% lactic acid was added to Chamber A (without adding citric acid), and the pH was adjusted to 5.5;

[0176] The freeze-drying process in Chamber B was the same as that in Example 3 (pre-freezing at -38°C and sublimation at 22°C);

[0177] The metal ion chelation index was not detected after reconstitution.

[0178] Comparison point: Lack of the chelating effect of citric acid to verify the synergistic contribution of the dual-acid system to oxidation stability.

[0179] Comparative Example 4 (non-vacuum homogenization):

[0180] Corresponding to Example 4: The vacuum environment was cancelled, and homogenization and mixing were carried out under normal pressure.

[0181] Preparation process:

[0182] The sub-packaging of Chambers A / B was the same as that in Example 4;

[0183] The aqueous phase and the oil phase were homogenized under normal pressure (pressure 10 MPa / 20 minutes), and dissolved oxygen was not removed;

[0184] Nitrogen protection was not used during filling.

[0185] Comparison point: Residual oxygen leads to accelerated oxidation to verify the inhibitory effect of vacuum homogenization and inert gas on oil rancidity.

[0186] Comparative Example 5 (premixing of active ingredients):

[0187] Corresponding to Example 5: Lutein and astaxanthin were directly added to the liquid system in Chamber A.

[0188] Preparation process:

[0189] 2% lutein and 1% astaxanthin were directly added to the liquid in Chamber A (not liposome-encapsulated);

[0190] Chamber B only contained 9% microcrystalline cellulose and 1% β-glucan;

[0191] No freeze-drying treatment was carried out, and the whole formula was stored in liquid form.

[0192] Comparison point: Active ingredients are exposed to light / oxygen environment to verify the improvement effect of liposome encapsulation and freeze-drying sub-packaging on photothermal stability.

[0193] Experiment 1:

[0194] Experimental procedures:

[0195] Sample preparation:

[0196] Example 1 group: Prepare 5 portions of A / B compartment separated samples according to the process of Example 1 (batch numbers E1-01 to E1-05);

[0197] Control 1 group: Prepare 5 portions of single compartment mixed samples according to the process of Control 1 (batch numbers C1-01 to C1-05).

[0198] Accelerated storage:

[0199] All samples were placed in a constant temperature and humidity chamber at 40°C / 75% RH and stored in the dark.

[0200] Sampling and detection:

[0201] Samples of each group were taken at 0, 1, 3, and 6 months respectively and detected by HPLC:

[0202] The purity of conotoxin (peak area normalization method, C18 chromatographic column, mobile phase acetonitrile - 0.1% TFA aqueous solution);

[0203] The content of ergothioneine (external standard method, detection wavelength 254 nm).

[0204] Data recording:

[0205] Record the retention rate (%) of active ingredients at each time point and calculate the degradation rate constant (k).

[0206] Experimental data:

[0207] Table 1: Influence of double-compartment isolation on the retention rate of active ingredients (40°C / 75% RH)

[0208]

[0209]

[0210] The physical isolation of the double compartment cut off the contact path between conotoxin and polyols (glycerol, butanediol). In the liquid environment, the amidation reaction induced by free hydroxyl groups was completely blocked. After 6 months of mixed storage in Control 1, the α-helical structure of conotoxin collapsed due to side chain modification, and unknown impurity peaks (retention time 2.8 - 3.1 minutes) appeared in the HPLC chromatogram, corresponding to more than 30% loss of activity.

[0211] The freeze-drying locking technology locks ergothioneine in a glassy matrix, and the water molecule mobility approaches zero. After the lyophilized powder of Example 1 is reconstituted, the liposomes are rapidly dispersed through micelle-mediated interfacial adsorption, avoiding the oxidative free radical chain reaction in traditional liquid storage. Due to the penetration of dissolved oxygen (initial oxygen concentration > 2 ppm) in the ergothioneine of Comparative Example 1, the generation amount of quinone derivatives reaches 28% after 6 months, and the ORAC value decays to 67% of the initial value.

[0212] The nitrogen laminar flow filling further suppresses the oxygen residue below the 0.1 ppm threshold. The vacuum gradient homogenization process of Example 1 makes the bubble diameter < 10 μm, avoiding the encapsulation and erosion of active substances by oxygen microbubbles. While the atmospheric pressure filling of Comparative Example 1 results in a gas content > 0.3% in the paste, and the oxidation reaction rate constant (k) during the acceleration period increases by 4.7 times, and the efficacy collapse occurs earlier than the industry standard time limit.

[0213] Experiment 2:

[0214] Experimental procedure:

[0215] Preparation of lyophilized powder:

[0216] Group of Example 2: Prepare 5 portions of lyophilized powder in Chamber B (batch numbers E2-01 to E2-05) according to the process of Example 2 (sublimation temperature 20 - 25 °C);

[0217] Group of Comparative Example 2: Prepare 5 portions of lyophilized powder in Chamber B (batch numbers C2-01 to C2-05) according to the process of Comparative Example 2 (sublimation temperature 32 °C).

[0218] Sample treatment:

[0219] All lyophilized powders are reconstituted in deionized water at 25 °C, and stirred at a speed of 400 rpm for 1 minute.

[0220] Structure observation:

[0221] Drop the reconstituted solution onto a copper mesh, negatively stain with phosphotungstic acid, and observe the liposome membrane morphology with a transmission electron microscope (TEM, Hitachi HT7800) (acceleration voltage 100 kV).

[0222] Transdermal test:

[0223] Ex vivo porcine skin (thickness 0.8 mm) is fixed in a Franz diffusion cell, and the receptor fluid is PBS at pH 7.4, and placed in a water bath at 32 °C;

[0224] Sampling time points: 2 h, 6 h, 12 h, 24 h, and HPLC is used to detect the cumulative permeation amount of ergothioneine (μg / cm 2 )

[0225] Experimental data:

[0226] Table 2: Influence of freeze-drying sublimation temperature on the transdermal performance of liposomes

[0227]

[0228] The low-temperature sublimation process suppresses the ice crystal interface temperature (Tice) below -20 °C. The phase transition temperature of the liposome bilayer membrane (Tm = 55 °C) is much higher than the sublimation environment, and the alkyl chains at the tails of phospholipid molecules maintain an all-trans conformation. The sublimation at 32 °C in Comparative Example 2 results in a local temperature exceeding Tm, a sharp increase in membrane fluidity, and liposome fusion (diameter > 200 nm) and membrane perforation (average breakage rate of 22%) are observed by electron microscopy.

[0229] Micelle-mediated dispersion depends on the integrity of the liposome structure. After reconstitution of the lyophilized powder in Example 2, emulsifier micelles (particle size 15 nm) are anchored to the liposome surface through hydrophobic interactions, promoting their embedding into the stratum corneum lipid matrix. Due to the leakage of membrane charge in the damaged liposomes in Comparative Example 2 (Zeta potential rising from -35 mV to -12 mV), the transdermal amount drops sharply to 0.11 μg / cm 2 , only 28% of that in the example group.

[0230] Gradient vacuum control avoids the remelting of ice crystals. The vacuum degree in the sublimation stage of Example 2 is ≤ 10 Pa, and ice crystals directly vaporize, skipping the liquid transition. The process defect in Comparative Example 2 causes partial melting of ice crystals, a sharp increase in solute concentration, resulting in an osmotic pressure difference, the breaking of the mannitol hydrogen bond network, and the aggregation of liposomes after reconstitution (PDI > 0.4), and the efficacy loss is irreversible.

[0231] Experiment 3:

[0232] Experimental procedure:

[0233] Sample treatment:

[0234] Example 3 group: Prepare 5 portions of the liquid in Chamber A (lactic acid + citric acid) according to the process of Example 3 (batch numbers E3-01 to E3-05);

[0235] Comparative Example 3 group: Prepare 5 portions of the liquid in Chamber A (only lactic acid) according to the process of Comparative Example 3 (batch numbers C3-01 to C3-05);

[0236] Add 0.1 ppm FeCl3 solution to all samples to simulate metal ion contamination.

[0237] Accelerated oxidation:

[0238] Place the samples of each group in an incubator at 40 °C and store them in the dark for 30 days.

[0239] Detection indexes:

[0240] ORAC value: Fluorescent probe method (excitation 485 nm / emission 520 nm), antioxidant capacity calculated by Trolox equivalent;

[0241] Metal Residue: Quantitative Analysis of Fe by ICP-MS (Agilent 7900) 3+ and Cu 2+ Free Ion Concentration

[0242] pH Stability: The pH meter (METTLER TOLEDO) records the fluctuation values before and after storage.

[0243] Experimental Data

[0244] Table 3: Regulation of Oxidation Stability by the Dual-Acid System

[0245]

[0246]

[0247] The tricarboxylic acid chelating group of citric acid locks free Fe 30 in the six-membered ring complex. The catalytic activity of metal ions is passivated, and the Fenton reaction chain process is directly interrupted. The single lactic acid system in Comparative Example 3 cannot capture Fe 3+ , and the ORAC value drops by nearly 40% within 30 days, and the concentration of the hydroxyl radical marker (8-OHdG) in the HPLC chromatogram soars 12-fold.

[0248] The α-hydroxy carboxylic acid structure of lactic acid provides a dynamic hydrogen bond network. The pH value fluctuation of the system is suppressed within the range of ±0.2, avoiding liposome membrane protonation caused by acid-base oscillation. The pH value of Comparative Example 3 drifts by more than 0.4, and the ζ potential on the membrane surface reverses from -32 mV to +5 mV, and the liposomes aggregate due to charge repulsion (particle size > 500 nm).

[0249] The dual-acid buffer gradient covers the pKa range of 3.1 - 6.4. The citric acid in Example 3 preferentially chelates high-valent metals, while lactic acid stabilizes the pH and penetrates the stratum corneum. The unchelated Cu in Comparative Example 3 2+ activates tyrosinase activity, accelerates the oxidation of ubiquinone in the formula (the production amount of quinone derivatives reaches 15%), and the efficacy shelf life is shortened to 3 months.

[0250] Experiment 4

[0251] Experimental Procedures

[0252] Sample Grouping

[0253] Example 4 Group: Prepare 5 portions of anti-acne and oil-control facial masks (batch numbers E4-01 to E4-05) according to the process of Example 4 (vacuum homogenization + nitrogen filling);

[0254] Comparative Example 4 Group: Prepare 5 portions of samples with the same formula (batch numbers C4-01 to C4-05) according to the process of Comparative Example 4 (atmospheric pressure homogenization).

[0255] Long-term storage:

[0256] All samples are stored in a dark environment at 25°C with a relative humidity ≤ 60%.

[0257] Index detection:

[0258] Peroxide value (PV): Samples are taken monthly, and the degree of oxidation of squalane is determined by the titration method according to GB5009.227-2016;

[0259] Microbial challenge: Staphylococcus aureus (ATCC6538) and Aspergillus niger (ATCC16404) are inoculated and cultured at 37°C for 7 days, and the colony count (CFU / g) is recorded;

[0260] Oxygen content: The headspace oxygen concentration in the package is monitored in real time by a laser oxygen sensor (PreSens Fibox4).

[0261] Experimental data:

[0262] Table 4: Inhibitory effects of vacuum homogenization on oil oxidation and microbial growth

[0263]

[0264]

[0265] Vacuum gradient homogenization reduces the dissolved oxygen to below 0.1 ppm. The concentration of oxygen free radicals around the double bond (C10-C11) of squalane molecules approaches zero, and the chain propagation of the peroxidation reaction is physically blocked. The residual oxygen in the atmospheric pressure process of Comparative Example 4 is > 2 ppm, and the accumulation amount of the primary product of oil oxidation (hydroperoxide) exceeds 5 meq / kg within 6 months, triggering the acid rancidity odor threshold in advance.

[0266] Nitrogen instantaneous filling forms an inert gas barrier layer. The headspace oxygen concentration in Example 4 is < 0.1%, and the metabolic pathway of aerobic bacteria (Staphylococcus aureus) stagnates due to lack of oxygen. The open filling in Comparative Example 4 results in an oxygen permeability > 2%, and microorganisms use glycerol as a carbon source, causing the colony count to soar to 10 3 grades, and the anti-corrosion system (phenoxyethanol) fails in advance.

[0267] The microbubble removal technology avoids the local enrichment of oxygen. The homogenization pressure (12 MPa) in Example 4 compresses the bubble diameter to < 5 μm, and the liposomes are evenly dispersed without oxygen hiding points. The process defect in Comparative Example 4 causes oxygen microbubbles (diameter > 50 μm) to be embedded in the paste, and the PV value in the oxidation hot spot area locally soars to 8 meq / kg, resulting in a complete collapse of stability.

[0268] Experiment 5:

[0269] Experimental procedure:

[0270] Sample exposure:

[0271] Example 5 group: Prepare 5 portions of anti-blue light repair cream according to the process of Example 5 (batch numbers E5-01 to E5-05);

[0272] Control group 5: Prepare 5 portions of liquid samples without encapsulated active substances according to the process of Control Example 5 (batch numbers C5-01 to C5-05);

[0273] All samples were laid flat in a quartz dish (thickness 1 mm) and placed in an illumination incubator (4500 Lux white light LED, 40 °C).

[0274] Stability tracking:

[0275] Samples were taken every 7 days and detected after being refrigerated in the dark:

[0276] UV-Vis spectroscopy: Detect the absorbance of characteristic peaks of lutein (447 nm) and astaxanthin (470 nm);

[0277] HPLC analysis: Separate photodegradation products with a C30 chromatographic column (mobile phase methanol-isopropanol gradient).

[0278] Cell experiment:

[0279] Human keratinocytes (HaCaT) were inoculated in a 96-well plate and irradiated with blue light (wavelength 450 nm, irradiance 30 mW / cm 2 ) for 30 minutes;

[0280] The cell viability was determined by the CCK-8 method (absorbance at 450 nm).

[0281] Experimental data:

[0282] Table 5: Regulation of photothermal stability by liposome encapsulation

[0283]

[0284] The hydrophobic barrier of the liposome bilayer membrane blocked the penetration of 450 nm high-energy blue light. The conjugated polyene chain of lutein was encapsulated inside the liposome, and the singlet oxygen ( 1 O2) quenching efficiency was increased to 93%. In the control group 5 where lutein was not encapsulated and directly exposed to the light environment, the proportion of cis isomers exceeded 58% after 14 days, and the light protection ability was completely lost.

[0285] The glassy matrix of the freeze-drying process locked the thermal motion of the active substance molecules. The astaxanthin in Example 5 formed an amorphous solid during pre-freezing at -40 °C, and the decay rate of the free radical scavenging activity (ORAC value) was suppressed within 10%. In the liquid formulation of the control group 5, the conjugated double bonds of astaxanthin were broken due to thermal vibration, and the proportion of the quinone structure soared to 42%, halving the cell repair efficiency.

[0286] The physical shielding of the cellulose skeleton weakens the oxygen diffusion path. The microcrystalline cellulose of Example 5 forms a dense network film on the paste surface, and the oxygen permeability coefficient (PO2) is as low as 0.05 Barrer. In the unformed film formulation of Comparative Example 5, the photooxidation reaction rate constant (k) reaches 4.8 times that of the example group, and the efficacy collapse is 2 times earlier than that of the same type of products in the market.

[0287] Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An antioxidant radiance mask process combining conotoxin and ergothioneine, characterized in that Comprising the following components in parts by mass: Active ingredient composition: 0.01 - 0.1% of conotoxin liposome-encapsulated, 0.2 - 1.0% of ergothioneine liposome-encapsulated; Matrix components: 50 - 70% of water, 3 - 8% of glycerol, 2 - 5% of butanediol, 2 - 5% of niacinamide, 1 - 5% of squalane, 3 - 8% of Limnanthes alba seed oil; Emulsifiers: 0.5 - 2% of polyglyceryl-10 laurate, 0.5 - 1.5% of sorbitan polyether-30 tetraoleate, 0.1 - 0.5% of sodium polyacryloyldimethyl taurate; Stabilizers: 0.5 - 2% of 1,2-hexanediol, 0.1 - 0.5% of p-hydroxyacetophenone, 0.05 - 0.2% of octanoyl hydroxamic acid.

2. The antioxidant and radiance-enhancing facial mask process combining conotoxin and ergothioneine according to claim 1, characterized in that, The active ingredient composition further comprises at least one of the following components: Palmitoyl tripeptide-1: 0.01 - 0.1%; Superoxide dismutase: 500 - 5000 U / g; Fullerene: 0.05 - 0.3%.

3. The antioxidant and radiance-boosting facial mask process combining conotoxin and ergothioneine according to claim 1, characterized in that, The matrix components further comprise at least one of the following components: Trehalose: 1 - 3%; Jojoba seed oil: 1 - 3%; Grape seed oil: 0.5 - 2%; Sodium hyaluronate: 0.1 - 0.5%.

4. The antioxidant and radiance-boosting facial mask process combining conotoxin and ergothioneine according to claim 1, characterized in that, The liposomes of conotoxin liposome-encapsulated and ergothioneine liposome-encapsulated contain: Hydrogenated lecithin: 2 - 5%; Cholesterol: 0.5 - 1.5%; Mannitol: 1 - 3%.

5. The antioxidant and radiance-enhancing facial mask process combining conotoxin and ergothioneine according to claim 4, characterized in that, The liposomes have a particle size of 50 - 100 nm and are prepared by high-pressure microfluidization homogenization process, with a homogenization pressure of 15,000 - 20,000 psi and a circulation times of 3 - 5 times.

6. The antioxidant and radiance-enhancing facial mask process combining conotoxin and ergothioneine according to claim 1, wherein The preparation process of the facial mask comprises the following steps: Heating the aqueous phase components to 40 - 50 °C for dissolution, and adding the active ingredient composition after cooling to 25 - 30 °C; Heating the oil phase components (squalane, Limnanthes alba seed oil, jojoba seed oil, grape seed oil) to 40 - 50 °C and mixing evenly; Mixing the aqueous phase and the oil phase in a vacuum homogenizer, with a homogenization pressure of 10 - 15 MPa and a mixing time of 15 - 30 min; Using inert gas protection during the final product mixing and filling process.

7. A process for an antioxidant radiance mask combining conotoxin and ergothioneine according to claim 6, characterized in that, The vacuum degree of the vacuum homogenizer is -0.08 to -0.1 MPa, and the stirring speed is 500 - 1000 rpm.

8. The antioxidant and radiance-boosting facial mask process combining conotoxin and ergothioneine according to claim 6, characterized in that The facial mask uses a two-chamber packaging, including: Chamber A: Liquid components containing the aqueous phase matrix and emulsifiers; Chamber B: Lyophilized powder containing conotoxin liposome-encapsulated and ergothioneine liposome-encapsulated.

9. The antioxidant and radiance-enhancing facial mask process combining conotoxin and ergothioneine according to claim 8, characterized in that, The preparation conditions of the lyophilized powder are: Pre-freezing temperature: -40 °C to -30 °C, pre-freezing time: 2 - 4 hours; Sublimation temperature: 20 - 25 °C, sublimation time: 8 - 12 hours, vacuum degree ≤ 10 Pa; Reconstitution temperature of the lyophilized powder is 25 - 30 °C, and the stirring speed is 200 - 400 rpm.

10. The antioxidant and radiance-enhancing facial mask process combining conotoxin and ergothioneine according to claim 1, wherein The pH value of the facial mask is 5.5 - 6.0 and is adjusted by lactic acid or citric acid.

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