Thermal spring water microelement liposome and application thereof in improving skin barrier function

Abstract

The invention provides a thermal spring water microelement liposome and an application thereof in improving a skin barrier function, the thermal spring water microelement liposome comprises the following components: 24-27 mg of hydrolyzed collagen, 50-60 mg of hydrogenated phosphatidylcholine, 34-39 mg of dilauroyl phosphatidylcholine, 30-35 mg of ceramide, 55-60 mg of cholesterol, 2-4 mg of hyaluronic acid, 1-5 mg of a preservative, and the balance of thermal spring water added to 1000 mg. The thermal spring water microelement lipidosome disclosed by the invention has the functions of improving a skin barrier function, replenishing water and preserving moisture, locking water and preserving moisture at the same time, reducing percutaneous moisture loss and moistening skin, and has the effects of softening, moistening and resisting oxidation.

Description

Technical Field

[0001] The invention belongs to the technical field of skin care products, and specifically relates to a hot spring water trace element liposome and an application thereof in improving skin barrier function. Background Art

[0002] As we age and our living environment deteriorates, the metabolism of our skin gradually declines, the absorption of nutrients also declines, and water is easily lost. At the same time, various harmful substances retained on the surface of the skin will also cause the skin to gradually lose its original elasticity, and the barrier protection function will gradually decline, thus causing a series of skin problems such as dryness, roughness, peeling, and wrinkles. Therefore, the repair and maintenance of the skin barrier has become an important part of people's daily life.

[0003] In response to the increasing number of skin problems, a variety of skin repair products have appeared on the market. However, in order to achieve immediate results, some merchants have added some hormone ingredients to the products, causing users to have new skin problems, such as hormone-dependent dermatitis, etc. There are also quite a few products containing chemical substances, which are not only irritating to the skin, but also easily cause allergic phenomena such as skin redness and swelling, and also damage the skin's barrier system, reducing the skin's ability to retain water. With the improvement of people's living standards, consumers have higher and higher requirements for cosmetics. While requiring products to have skin repair effects, they also require products to be natural, safe, healthy and mild.

[0004] Therefore, there is an urgent need to develop a product that has good skin barrier repair effect and is safe and gentle to meet people's needs. Summary of the invention

[0005] The purpose of the invention is to provide a hot spring water trace element liposome and its application in improving skin barrier function.

[0006] In order to achieve the above object, the present invention provides the following technical solutions: A hot spring water trace element liposome comprises the following components: 24-27 mg of hydrolyzed collagen, 50-60 mg of hydrogenated phosphatidylcholine, 34-39 mg of dilauroyl phosphatidylcholine, 30-35 mg of ceramide, 55-60 mg of cholesterol, 2-4 mg of hyaluronic acid, 1-5 mg of preservative, and hot spring water added to 1000 mg.

[0007] Furthermore, each liter of the hot spring water contains the following components: 2.58-2.76 mg of K + 45.7-53.8 mg of Na + , 10.2-11.3 mg of Ca 2+ , 0.25-0.46 mg Mg 2+, 0.018-0.027 mg of Tfe total iron and 70.91-76.42 mg of metasilicic acid.

[0008] Furthermore, the preparation method of hydrolyzed collagen is as follows: S1: pretreating the cowhide to obtain pretreated cowhide; S2: homogenizing and digesting the pretreated cowhide to obtain a homogenized digestion solution; wherein the homogenizing and digesting comprises a homogenizing treatment and a digestion treatment using pepsin; S3: performing adsorption filtration on the homogenized digestion solution to obtain an adsorption filtrate; S4: sequentially performing concentration and liquid replacement, ion exchange chromatography, secondary concentration and liquid replacement, filtration and sterilization, and centrifugal purification on at least part of the adsorption filtrate to obtain collagen; S5: collecting the waste liquid from steps S1 to S4, using the collected waste liquid and / or another part of the adsorption filtrate as a pre-hydrolysis liquid, performing acid hydrolysis on the pre-hydrolysis liquid to obtain hydrolyzed collagen.

[0009] Further, in step S1, the pretreatment may include: washing and cleaning the cowhide, then soaking it in a 1-10wt% acetic acid solution, and crushing the soaked cowhide into cowhide particles to obtain pretreated cowhide; wherein the soaking temperature may be controlled to be 4-25°C, and the soaking time may be 5-10 days. Preferably, the concentration of the acetic acid solution for soaking is 5-10wt%.

[0010] Further, in step S2, the homogenization treatment may include: adding injection water to the pretreated cowhide, homogenizing the pretreated cowhide with a colloid mill, adjusting the pH value to 2.0-3.0 after homogenization, and obtaining a homogenate; wherein, the ratio of the pretreated cowhide to the injection water may be 1kg:(2-8)L; the colloid mill is pre-soaked in a 0.4-0.6M sodium hydroxide solution for not less than 30 minutes; glacial acetic acid may be used to adjust the pH value to 2.0-3.0; the cowhide content in the homogenate may be 20-25g / L.

[0011] Further, in step S3, the adsorption filtration may include: adjusting the pH value of the homogenized digestion solution to 3.9-4.1, then adding pretreated diatomaceous earth for static adsorption, collecting the supernatant and filtering to obtain an adsorption filtrate; wherein, the preparation method of the pretreated diatomaceous earth may include: soaking the diatomaceous earth in 8-12mM hydrochloric acid solution for more than 2h, and washing it with purified water for more than 3 times after soaking to obtain the pretreated diatomaceous earth; the amount of pretreated diatomaceous earth added may be 14-16g / L homogenized digestion solution; in addition, the static adsorption time may be controlled to be 1-3h.

[0012] Further, in step S4, the concentration and liquid exchange and the secondary concentration and liquid exchange respectively include: using a 100kD ultrafiltration membrane, using an acetate buffer of pH 4-5 to exchange the liquid 1-3 times, and concentrating and collecting the liquid after the liquid exchange.

[0013] Furthermore, in step S5, a 5.5-6.5 mol / L hydrochloric acid solution can be used for acid hydrolysis; the amount of hydrochloric acid solution used during acid hydrolysis is 1.5-2% of the volume of the pre-hydrolysis solution; in addition, the acid hydrolysis temperature can be controlled to be 55-70°C, and the acid hydrolysis time can be 40-48h.

[0014] Furthermore, the preparation method includes the following steps: dissolving phospholipids, ceramides and cholesterol in ether, taking hydrolyzed collagen separately, adding water to dissolve it, adding the ether solution containing phospholipids, ceramides and cholesterol to the hydrolyzed collagen aqueous solution, and subjecting it to short-term ultrasound or vibration to form a stable W / O type emulsion, and then removing the ether under reduced pressure evaporation conditions to obtain a gel-like substance, and then dripping 0.1M pH 6.0 phosphate buffer into the above substance, vortexing and vibrating to make the gel fall off, and continuing to evaporate under reduced pressure, and taking hyaluronic acid and preservatives separately, adding them to the solution to dissolve, and obtaining hot spring water trace element liposomes.

[0015] Application of hot spring water trace element liposomes in improving skin barrier function.

[0016] Compared with the prior art, the advantages and beneficial effects of the present invention are: 1. The hydrolyzed collagen of the present invention has a good moisturizing effect, can penetrate into the deep layer of the skin, increase the water content of the skin, and thus play a role in moisturizing. Collagen contains hydrophilic natural moisturizing factors, which can help the skin lock in moisture, prevent water loss, and keep the skin moist. Hydrolyzed collagen can stimulate skin cells to produce new collagen and elastic fibers, help repair damaged skin tissue, and promote skin cell renewal. By stimulating the synthesis of new collagen, the skin repair process is accelerated, inflammation and irritation are reduced, and the skin barrier is restored.

[0017] 2. In the hot spring water of the present invention, potassium ions are very important for maintaining the water balance inside and outside the skin cells. It can help the skin maintain an appropriate water level, thereby helping to maintain the integrity of the skin barrier. Sodium ions also play a role in maintaining the water balance of the skin. It can help maintain the osmotic pressure balance of the skin and prevent excessive evaporation of water, thereby helping to repair the skin barrier. Calcium ions are essential for the skin barrier function. It helps to maintain the connection between skin cells, enhances the tightness of the skin barrier, and also promotes the self-repair mechanism of the skin and reduces inflammatory reactions. Magnesium ions help to reduce skin inflammation and have a certain anti-inflammatory effect. In addition, magnesium ions can also help the skin maintain appropriate moisture, thereby helping to maintain the healthy state of the skin barrier. Iron is a component of many enzymes, participates in the metabolic process of the skin, and helps to maintain the normal function of the skin. Metasilicic acid is considered to have a moisturizing effect on the skin and can help improve the skin barrier function. It can also promote the self-repair of the skin and reduce inflammatory reactions, which is very beneficial for maintaining the healthy state of the skin. These trace elements work together to help repair damaged skin barriers in many ways, such as providing moisturizing, reducing inflammation, promoting skin cell renewal, and enhancing skin barrier function. DETAILED DESCRIPTION

[0018] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0019] Example 1 This embodiment provides a hot spring water trace element liposome, comprising the following components: 27 mg of hydrolyzed collagen, 60 mg of hydrogenated phosphatidylcholine, 39 mg of dilauroyl phosphatidylcholine, 35 mg of ceramide, 60 mg of cholesterol, 4 mg of hyaluronic acid, 5 mg of preservative, and hot spring water added to 1000 mg.

[0020] Each liter of the hot spring water contains the following components: 2.76 mg of K + , 53.8 mg of Na + , 11.3 mg of Ca 2+ , 0.46 mg Mg 2+ , 0.027 mg of Tfe total iron and 76.42 mg of metasilicic acid.

[0021] The preparation method of hydrolyzed collagen is as follows: S1: pretreating the cowhide to obtain pretreated cowhide; S2: homogenizing and digesting the pretreated cowhide to obtain a homogenized digestion solution; wherein the homogenizing and digesting comprises a homogenizing treatment and a digestion treatment using pepsin; S3: performing adsorption filtration on the homogenized digestion solution to obtain an adsorption filtrate; S4: sequentially performing concentration and liquid replacement, ion exchange chromatography, secondary concentration and liquid replacement, filtration and sterilization, and centrifugal purification on at least part of the adsorption filtrate to obtain collagen; S5: collecting the waste liquid from steps S1 to S4, using the collected waste liquid as a pre-hydrolysis liquid, and subjecting the pre-hydrolysis liquid to acid hydrolysis to obtain hydrolyzed collagen.

[0022] In step S1, the pretreatment may include: washing and cleaning the cowhide, then soaking it in a 10wt% acetic acid solution, and crushing the soaked cowhide into cowhide particles to obtain pretreated cowhide; wherein the soaking temperature may be controlled to be 25°C and the soaking time may be 10 days. Preferably, the concentration of the acetic acid solution for soaking is 10wt%.

[0023] In step S2, the homogenization treatment may include: adding injection water to the pretreated cowhide, homogenizing the pretreated cowhide with a colloid mill, adjusting the pH value to 3.0 after homogenization, and obtaining a homogenate; wherein, the ratio of the pretreated cowhide to the injection water may be 1kg: 8L; the colloid mill may be pre-soaked in 0.6M sodium hydroxide solution for 60 minutes; glacial acetic acid may be used to adjust the pH value to 3.0; the cowhide content in the homogenate may be 25g / L.

[0024] In step S3, the adsorption filtration may include: adjusting the pH value of the homogenized digestion solution to 4.1, then adding pretreated diatomaceous earth for static adsorption, collecting the supernatant and filtering it to obtain an adsorption filtrate; wherein, the preparation method of the pretreated diatomaceous earth may include: soaking the diatomaceous earth in a 2mM hydrochloric acid solution for 3 hours, and washing it with purified water for 5 times after soaking to obtain the pretreated diatomaceous earth; the amount of pretreated diatomaceous earth added may be 16g / L homogenized digestion solution; in addition, the static adsorption time may be controlled to be 3 hours.

[0025] In step S4, the concentration and liquid exchange and the secondary concentration and liquid exchange respectively include: using a 100 kD ultrafiltration membrane, using a pH 5 acetate buffer to exchange the liquid three times, and concentrating and collecting the liquid after the liquid exchange.

[0026] In step S5, a 6.5 mol / L hydrochloric acid solution can be used for acid hydrolysis; the amount of hydrochloric acid solution used during acid hydrolysis is 2% of the volume of the solution before hydrolysis; in addition, the acid hydrolysis temperature can be controlled to 70° C. and the acid hydrolysis time can be controlled to 48 hours.

[0027] The preparation method of hot spring water trace element liposomes comprises the following steps: dissolving phospholipids, ceramide and cholesterol in ether, separately taking hydrolyzed collagen, adding water to dissolve it, adding the ether solution containing phospholipids, ceramide and cholesterol into the hydrolyzed collagen aqueous solution, subjecting to short-time ultrasound or oscillation to form a stable W / O type emulsion, then removing the ether under reduced pressure evaporation conditions to obtain a gel-state substance, then dripping 0.1M pH 6.0 phosphate buffer into the above substance, vortexing and oscillating to make the gel fall off, continuing to evaporate under reduced pressure, separately taking hyaluronic acid and a preservative to add them into the solution to dissolve, and preparing hot spring water trace element liposomes.

[0028] Example 2 This embodiment provides a hot spring water trace element liposome, comprising the following components: 24 mg of hydrolyzed collagen, 50 mg of hydrogenated phosphatidylcholine, 34 mg of dilauroyl phosphatidylcholine, 30 mg of ceramide, 55 mg of cholesterol, 2 mg of hyaluronic acid, 1 mg of preservative, and hot spring water added to 1000 mg.

[0029] Each liter of the hot spring water contains the following components: 2.58 mg of K + 45.7 mg of Na + , 10.2 mg of Ca 2+ , 0.25 mg Mg 2+ , 0.018 mg of Tfe total iron and 70.91 mg of metasilicic acid.

[0030] The preparation method of hydrolyzed collagen is as follows: S1: pretreating the cowhide to obtain pretreated cowhide; S2: homogenizing and digesting the pretreated cowhide to obtain a homogenized digestion solution; wherein the homogenizing and digesting comprises a homogenizing treatment and a digestion treatment using pepsin; S3: performing adsorption filtration on the homogenized digestion solution to obtain an adsorption filtrate; S4: sequentially performing concentration and liquid replacement, ion exchange chromatography, secondary concentration and liquid replacement, filtration and sterilization, and centrifugal purification on at least part of the adsorption filtrate to obtain collagen; S5: collecting the waste liquid from steps S1 to S4, using the collected waste liquid as a pre-hydrolysis liquid, and subjecting the pre-hydrolysis liquid to acid hydrolysis to obtain hydrolyzed collagen.

[0031] In step S1, the pretreatment may include: washing and cleaning the cowhide, then soaking it in a 1wt% acetic acid solution, and crushing the soaked cowhide into cowhide particles to obtain pretreated cowhide; wherein the soaking temperature may be controlled to be 4°C and the soaking time may be 5 days. Preferably, the concentration of the acetic acid solution used for soaking is 5wt%.

[0032] In step S2, the homogenization treatment may include: adding water for injection to the pretreated cowhide, homogenizing the pretreated cowhide with a colloid mill, adjusting the pH value to 2.0 after homogenization, and obtaining a homogenate; wherein, the ratio of the pretreated cowhide to the water for injection may be 1kg:2L; the colloid mill may be pre-soaked in 0.4 sodium hydroxide solution for 40 minutes; glacial acetic acid may be used to adjust the pH value to 2.0; the cowhide content in the homogenate may be 20g / L.

[0033] In step S3, the adsorption filtration may include: adjusting the pH value of the homogenized digestion solution to 3.9, then adding pretreated diatomaceous earth for static adsorption, collecting the supernatant and filtering it to obtain an adsorption filtrate; wherein, the preparation method of the pretreated diatomaceous earth may include: soaking the diatomaceous earth in 8mM hydrochloric acid solution for 3h, and washing it with purified water 4 times after soaking to obtain the pretreated diatomaceous earth; the amount of pretreated diatomaceous earth added may be 14g / L homogenized digestion solution; in addition, the static adsorption time may be controlled to be 1h.

[0034] In step S4, the concentration and liquid exchange and the secondary concentration and liquid exchange respectively include: using a 100 kD ultrafiltration membrane, using a pH 4 acetate buffer to exchange the liquid once, and concentrating and collecting the liquid after the liquid exchange.

[0035] In step S5, a 5.5 mol / L hydrochloric acid solution can be used for acid hydrolysis; the amount of hydrochloric acid solution used during acid hydrolysis is 1.5% of the volume of the solution before hydrolysis; in addition, the acid hydrolysis temperature can be controlled to be 55° C. and the acid hydrolysis time can be controlled to be 40 h.

[0036] The preparation method of hot spring water trace element liposomes comprises the following steps: dissolving phospholipids, ceramide and cholesterol in ether, separately taking hydrolyzed collagen, adding water to dissolve it, adding the ether solution containing phospholipids, ceramide and cholesterol into the hydrolyzed collagen aqueous solution, subjecting to short-time ultrasound or oscillation to form a stable W / O type emulsion, then removing the ether under reduced pressure evaporation conditions to obtain a gel-state substance, then dripping 0.1M pH 6.0 phosphate buffer into the above substance, vortexing and oscillating to make the gel fall off, continuing to evaporate under reduced pressure, separately taking hyaluronic acid and a preservative to add them into the solution to dissolve, and preparing hot spring water trace element liposomes.

[0037] Comparative Example 1 The difference between this comparative example and Example 1 is: a hot spring water trace element liposome, comprising the following components: 14 mg of hydrolyzed collagen, 50 mg of hydrogenated phosphatidylcholine, 34 mg of dilauroyl phosphatidylcholine, 40 mg of ceramide, 55 mg of cholesterol, 2 mg of hyaluronic acid, 1 mg of preservative, and hot spring water added to 1000 mg.

[0038] Specifically: A hot spring water trace element liposome comprises the following components: 14 mg of hydrolyzed collagen, 50 mg of hydrogenated phosphatidylcholine, 34 mg of dilauroyl phosphatidylcholine, 40 mg of ceramide, 55 mg of cholesterol, 2 mg of hyaluronic acid, 1 mg of preservative, and hot spring water added to 1000 mg.

[0039] Each liter of the hot spring water contains the following components: 2.76 mg of K + , 53.8 mg of Na + , 11.3 mg of Ca 2+ , 0.46 mg Mg 2+ , 0.027 mg of Tfe total iron and 76.42 mg of metasilicic acid.

[0040] The preparation method of hydrolyzed collagen is as follows: S1: pretreating the cowhide to obtain pretreated cowhide; S2: homogenizing and digesting the pretreated cowhide to obtain a homogenized digestion solution; wherein the homogenizing and digesting comprises a homogenizing treatment and a digestion treatment using pepsin; S3: performing adsorption filtration on the homogenized digestion solution to obtain an adsorption filtrate; S4: sequentially performing concentration and liquid replacement, ion exchange chromatography, secondary concentration and liquid replacement, filtration and sterilization, and centrifugal purification on at least part of the adsorption filtrate to obtain collagen; S5: collecting the waste liquid from steps S1 to S4, using the collected waste liquid as a pre-hydrolysis liquid, and subjecting the pre-hydrolysis liquid to acid hydrolysis to obtain hydrolyzed collagen.

[0041] In step S1, the pretreatment may include: washing and cleaning the cowhide, then soaking it in a 10wt% acetic acid solution, and crushing the soaked cowhide into cowhide particles to obtain pretreated cowhide; wherein the soaking temperature may be controlled to be 25°C and the soaking time may be 10 days. Preferably, the concentration of the acetic acid solution for soaking is 10wt%.

[0042] In step S2, the homogenization treatment may include: adding injection water to the pretreated cowhide, homogenizing the pretreated cowhide with a colloid mill, adjusting the pH value to 3.0 after homogenization, and obtaining a homogenate; wherein, the ratio of the pretreated cowhide to the injection water may be 1kg: 8L; the colloid mill may be pre-soaked in 0.6M sodium hydroxide solution for 60 minutes; glacial acetic acid may be used to adjust the pH value to 3.0; the cowhide content in the homogenate may be 25g / L.

[0043] In step S3, the adsorption filtration may include: adjusting the pH value of the homogenized digestion solution to 4.1, then adding pretreated diatomaceous earth for static adsorption, collecting the supernatant and filtering it to obtain an adsorption filtrate; wherein, the preparation method of the pretreated diatomaceous earth may include: soaking the diatomaceous earth in a 2mM hydrochloric acid solution for 3 hours, and washing it with purified water for 5 times after soaking to obtain the pretreated diatomaceous earth; the amount of pretreated diatomaceous earth added may be 16g / L homogenized digestion solution; in addition, the static adsorption time may be controlled to be 3 hours.

[0044] In step S4, the concentration and liquid exchange and the secondary concentration and liquid exchange respectively include: using a 100 kD ultrafiltration membrane, using a pH 5 acetate buffer to exchange the liquid three times, and concentrating and collecting the liquid after the liquid exchange.

[0045] In step S5, a 6.5 mol / L hydrochloric acid solution can be used for acid hydrolysis; the amount of hydrochloric acid solution used during acid hydrolysis is 2% of the volume of the solution before hydrolysis; in addition, the acid hydrolysis temperature can be controlled to 70° C. and the acid hydrolysis time can be controlled to 48 hours.

[0046] The preparation method of hot spring water trace element liposomes comprises the following steps: dissolving phospholipids, ceramide and cholesterol in ether, separately taking hydrolyzed collagen, adding water to dissolve it, adding the ether solution containing phospholipids, ceramide and cholesterol into the hydrolyzed collagen aqueous solution, subjecting to short-time ultrasound or oscillation to form a stable W / O type emulsion, then removing the ether under reduced pressure evaporation conditions to obtain a gel-state substance, then dripping 0.1M pH 6.0 phosphate buffer into the above substance, vortexing and oscillating to make the gel fall off, continuing to evaporate under reduced pressure, separately taking hyaluronic acid and a preservative to add them into the solution to dissolve, and preparing hot spring water trace element liposomes.

[0047] Comparative Example 2 The difference between this comparative example and Example 1 is that each liter of the hot spring water contains the following components: 1.58 mg of K + , 46.7 mg of Na + , 9.2 mg of Ca 2+ , 0.35 mg Mg 2+ , 0.008 mg of Tfe total iron and 71.91 mg of metasilicic acid.

[0048] Specifically: A hot spring water trace element liposome comprises the following components: 27 mg of hydrolyzed collagen, 60 mg of hydrogenated phosphatidylcholine, 39 mg of dilauroyl phosphatidylcholine, 35 mg of ceramide, 60 mg of cholesterol, 4 mg of hyaluronic acid, 5 mg of preservative, and hot spring water added to 1000 mg.

[0049] Each liter of the hot spring water contains the following components: 1.58 mg of K + , 46.7 mg of Na+ , 9.2 mg of Ca 2+ , 0.35 mg Mg 2+ , 0.008 mg of Tfe total iron and 71.91 mg of metasilicic acid.

[0050] The preparation method of hydrolyzed collagen is as follows: S1: pretreating the cowhide to obtain pretreated cowhide; S2: homogenizing and digesting the pretreated cowhide to obtain a homogenized digestion solution; wherein the homogenizing and digesting comprises a homogenizing treatment and a digestion treatment using pepsin; S3: performing adsorption filtration on the homogenized digestion solution to obtain an adsorption filtrate; S4: sequentially performing concentration and liquid replacement, ion exchange chromatography, secondary concentration and liquid replacement, filtration and sterilization, and centrifugal purification on at least part of the adsorption filtrate to obtain collagen; S5: collecting the waste liquid from steps S1 to S4, using the collected waste liquid as a pre-hydrolysis liquid, and subjecting the pre-hydrolysis liquid to acid hydrolysis to obtain hydrolyzed collagen.

[0051] In step S1, the pretreatment may include: washing and cleaning the cowhide, then soaking it in a 10wt% acetic acid solution, and crushing the soaked cowhide into cowhide particles to obtain pretreated cowhide; wherein the soaking temperature may be controlled to be 25°C and the soaking time may be 10 days. Preferably, the concentration of the acetic acid solution for soaking is 10wt%.

[0052] In step S2, the homogenization treatment may include: adding injection water to the pretreated cowhide, homogenizing the pretreated cowhide with a colloid mill, adjusting the pH value to 3.0 after homogenization, and obtaining a homogenate; wherein, the ratio of the pretreated cowhide to the injection water may be 1kg: 8L; the colloid mill may be pre-soaked in 0.6M sodium hydroxide solution for 60 minutes; glacial acetic acid may be used to adjust the pH value to 3.0; the cowhide content in the homogenate may be 25g / L.

[0053] In step S3, the adsorption filtration may include: adjusting the pH value of the homogenized digestion solution to 4.1, then adding pretreated diatomaceous earth for static adsorption, collecting the supernatant and filtering it to obtain an adsorption filtrate; wherein, the preparation method of the pretreated diatomaceous earth may include: soaking the diatomaceous earth in a 2mM hydrochloric acid solution for 3 hours, and washing it with purified water for 5 times after soaking to obtain the pretreated diatomaceous earth; the amount of pretreated diatomaceous earth added may be 16g / L homogenized digestion solution; in addition, the static adsorption time may be controlled to be 3 hours.

[0054] In step S4, the concentration and liquid exchange and the secondary concentration and liquid exchange respectively include: using a 100 kD ultrafiltration membrane, using a pH 5 acetate buffer to exchange the liquid three times, and concentrating and collecting the liquid after the liquid exchange.

[0055] In step S5, a 6.5 mol / L hydrochloric acid solution can be used for acid hydrolysis; the amount of hydrochloric acid solution used during acid hydrolysis is 2% of the volume of the solution before hydrolysis; in addition, the acid hydrolysis temperature can be controlled to 70° C. and the acid hydrolysis time can be controlled to 48 hours.

[0056] The preparation method of hot spring water trace element liposomes comprises the following steps: dissolving phospholipids, ceramide and cholesterol in ether, separately taking hydrolyzed collagen, adding water to dissolve it, adding the ether solution containing phospholipids, ceramide and cholesterol into the hydrolyzed collagen aqueous solution, subjecting to short-time ultrasound or oscillation to form a stable W / O type emulsion, then removing the ether under reduced pressure evaporation conditions to obtain a gel-state substance, then dripping 0.1M pH 6.0 phosphate buffer into the above substance, vortexing and oscillating to make the gel fall off, continuing to evaporate under reduced pressure, separately taking hyaluronic acid and a preservative to add them into the solution to dissolve, and preparing hot spring water trace element liposomes.

[0057] Comparative Example 3 The difference between this comparative example and Example 1 is: S2: homogenizing and digesting the pretreated cowhide to obtain a homogenized digestion solution; wherein the homogenized digestion includes a homogenization treatment and a digestion treatment using trypsin.

[0058] Specifically: A hot spring water trace element liposome comprises the following components: 27 mg of hydrolyzed collagen, 60 mg of hydrogenated phosphatidylcholine, 39 mg of dilauroyl phosphatidylcholine, 35 mg of ceramide, 60 mg of cholesterol, 4 mg of hyaluronic acid, 5 mg of preservative, and hot spring water added to 1000 mg.

[0059] Each liter of the hot spring water contains the following components: 2.76 mg of K + , 53.8 mg of Na + , 11.3 mg of Ca 2+ , 0.46 mg Mg 2+ , 0.027 mg of Tfe total iron and 76.42 mg of metasilicic acid.

[0060] The preparation method of hydrolyzed collagen is as follows: S1: pretreating the cowhide to obtain pretreated cowhide; S2: homogenizing and digesting the pretreated cowhide to obtain a homogenized digestion solution; wherein the homogenizing and digesting comprises a homogenizing treatment and a digestion treatment using trypsin; S3: performing adsorption filtration on the homogenized digestion solution to obtain an adsorption filtrate; S4: sequentially performing concentration and liquid replacement, ion exchange chromatography, secondary concentration and liquid replacement, filtration and sterilization, and centrifugal purification on at least part of the adsorption filtrate to obtain collagen; S5: collecting the waste liquid from steps S1 to S4, using the collected waste liquid as a pre-hydrolysis liquid, and subjecting the pre-hydrolysis liquid to acid hydrolysis to obtain hydrolyzed collagen.

[0061] In step S1, the pretreatment may include: washing and cleaning the cowhide, then soaking it in a 10wt% acetic acid solution, and crushing the soaked cowhide into cowhide particles to obtain pretreated cowhide; wherein the soaking temperature may be controlled to be 25°C and the soaking time may be 10 days. Preferably, the concentration of the acetic acid solution for soaking is 10wt%.

[0062] In step S2, the homogenization treatment may include: adding injection water to the pretreated cowhide, homogenizing the pretreated cowhide with a colloid mill, adjusting the pH value to 3.0 after homogenization, and obtaining a homogenate; wherein, the ratio of the pretreated cowhide to the injection water may be 1kg: 8L; the colloid mill may be pre-soaked in 0.6M sodium hydroxide solution for 60 minutes; glacial acetic acid may be used to adjust the pH value to 3.0; the cowhide content in the homogenate may be 25g / L.

[0063] In step S3, the adsorption filtration may include: adjusting the pH value of the homogenized digestion solution to 4.1, then adding pretreated diatomaceous earth for static adsorption, collecting the supernatant and filtering it to obtain an adsorption filtrate; wherein, the preparation method of the pretreated diatomaceous earth may include: soaking the diatomaceous earth in a 2mM hydrochloric acid solution for 3 hours, and washing it with purified water for 5 times after soaking to obtain the pretreated diatomaceous earth; the amount of pretreated diatomaceous earth added may be 16g / L homogenized digestion solution; in addition, the static adsorption time may be controlled to be 3 hours.

[0064] In step S4, the concentration and liquid exchange and the secondary concentration and liquid exchange respectively include: using a 100 kD ultrafiltration membrane, using a pH 5 acetate buffer to exchange the liquid three times, and concentrating and collecting the liquid after the liquid exchange.

[0065] In step S5, a 6.5 mol / L hydrochloric acid solution can be used for acid hydrolysis; the amount of hydrochloric acid solution used during acid hydrolysis is 2% of the volume of the solution before hydrolysis; in addition, the acid hydrolysis temperature can be controlled to 70° C. and the acid hydrolysis time can be controlled to 48 hours.

[0066] The preparation method of hot spring water trace element liposomes comprises the following steps: dissolving phospholipids, ceramide and cholesterol in ether, separately taking hydrolyzed collagen, adding water to dissolve it, adding the ether solution containing phospholipids, ceramide and cholesterol into the hydrolyzed collagen aqueous solution, subjecting to short-time ultrasound or oscillation to form a stable W / O type emulsion, then removing the ether under reduced pressure evaporation conditions to obtain a gel-state substance, then dripping 0.1M pH 6.0 phosphate buffer into the above substance, vortexing and oscillating to make the gel fall off, continuing to evaporate under reduced pressure, separately taking hyaluronic acid and a preservative to add them into the solution to dissolve, and preparing hot spring water trace element liposomes.

[0067] Comparative Example 4 The difference between this comparative example and Example 1 is that the hydrolyzed collagen is a commercially available product.

[0068] A hot spring water trace element liposome comprises the following components: 27 mg of hydrolyzed collagen, 60 mg of hydrogenated phosphatidylcholine, 39 mg of dilauroyl phosphatidylcholine, 35 mg of ceramide, 60 mg of cholesterol, 4 mg of hyaluronic acid, 5 mg of preservative, and hot spring water added to 1000 mg.

[0069] Each liter of the hot spring water contains the following components: 2.76 mg of K + , 53.8 mg of Na + , 11.3 mg of Ca 2+ , 0.46 mg Mg 2+ , 0.027 mg of Tfe total iron and 76.42 mg of metasilicic acid.

[0070] The preparation method of hot spring water trace element liposomes comprises the following steps: dissolving phospholipids, ceramide and cholesterol in ether, separately taking hydrolyzed collagen, adding water to dissolve it, adding the ether solution containing phospholipids, ceramide and cholesterol into the hydrolyzed collagen aqueous solution, subjecting to short-time ultrasound or oscillation to form a stable W / O type emulsion, then removing the ether under reduced pressure evaporation conditions to obtain a gel-state substance, then dripping 0.1M pH 6.0 phosphate buffer into the above substance, vortexing and oscillating to make the gel fall off, continuing to evaporate under reduced pressure, separately taking hyaluronic acid and a preservative to add them into the solution to dissolve, and preparing hot spring water trace element liposomes.

[0071] Performance Testing 60 volunteers were selected as subjects, aged 20-45 years old, with healthy skin, no history of skin allergies, and met the voluntary inclusion criteria of the subjects. They were randomly divided into 6 groups for testing, and each group used a sample. The hot spring water extracts of Examples 1-2 and Comparative Examples 1-4 were used as samples. The subjects applied an equal amount of the sample (2 mL g) evenly on the inner side of the forearm twice a day, morning and evening, and then the same tester used the TM300 tester of the German CK company to test the transepidermal water loss of the inner side of the forearm in the 0th week and after continuous use until the 3rd week, abbreviated as TEWL, that is, how many grams of water are lost per square meter per hour.

[0072] TEWL is a very important parameter for evaluating the function of the skin's moisture barrier. It has been widely recognized internationally and can well reflect the integrity of the skin's barrier function. The standard TEWL is measured when the skin is dry and sweat-free, at an ambient temperature of 24°C and an ambient humidity of 50%. The test data are all obtained under these conditions.

[0073] The calculation formula of TEWL value decrease rate is as follows: TEWL value reduction rate (%) = (A0-An) / A0×100% Where A0 is the TEWL value of the 0th week, and An is the TEWL value of the nth week.

[0074] A positive TEWL value decrease rate indicates that the skin barrier has been repaired, otherwise, the skin barrier has not been repaired; and the greater the TEWL value decrease rate, the better the skin barrier repair effect. The calculation formula for the average TEWL value decrease rate is as follows: Average TEWL value reduction rate (%) = sum of TEWL value reduction rates of each group / number of people in the group The test results are shown in Table 1.

[0075] Table 1 Performance test results

[0076] From the above performance test results, it can be seen that Example 1 has outstanding skin barrier repair performance. However, since the comparative example does not adopt the necessary technical solution, it is obviously worse than the example in the corresponding performance test. The above experimental results further prove the importance of the technical solution defined in the present invention for its technical effect.

[0077] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A hot spring water trace element liposome, characterized in that: It includes the following ingredients: hydrolyzed collagen 24-27mg, hydrogenated phosphatidylcholine 50-60mg, dilauroyl phosphatidylcholine 34-39mg, ceramide 30-35mg, cholesterol 55-60mg, hyaluronic acid 2-4mg, preservative 1-5mg, and hot spring water added to 1000mg.

2. The hot spring water trace element liposome according to claim, characterized in that: Each liter of the hot spring water contains the following components: 2.58-2.76 mg of K + 45.7-53.8 mg of Na + , 10.2-11.3 mg of Ca 2+ , 0.25-0.46 mg Mg 2+ , 0.018-0.027 mg of Tfe total iron and 70.91-76.42 mg of metasilicic acid.

3. The hot spring water trace element liposome according to claim, characterized in that: The preparation method of hydrolyzed collagen is as follows: S1: pretreating the cowhide to obtain pretreated cowhide; S2: homogenizing and digesting the pretreated cowhide to obtain a homogenized digestion solution; wherein the homogenizing and digesting comprises a homogenizing treatment and a digestion treatment using pepsin; S3: performing adsorption filtration on the homogenized digestion solution to obtain an adsorption filtrate; S4: sequentially performing concentration and liquid replacement, ion exchange chromatography, secondary concentration and liquid replacement, filtration and sterilization, and centrifugal purification on at least part of the adsorption filtrate to obtain collagen; S5: collecting the waste liquid from steps S1 to S4, using the collected waste liquid and / or another part of the adsorption filtrate as a pre-hydrolysis liquid, performing acid hydrolysis on the pre-hydrolysis liquid to obtain hydrolyzed collagen.

4. The hot spring water trace element liposome according to claim 2, characterized in that: In step S1, the pretreatment may include: washing and cleaning the cowhide, then soaking it in a 1-10wt% acetic acid solution, and crushing the soaked cowhide into cowhide particles to obtain pretreated cowhide; wherein the soaking temperature may be controlled to be 4-25°C, and the soaking time may be 5-10 days. Preferably, the concentration of the acetic acid solution for soaking is 5-10wt%.

5. The hot spring water trace element liposome according to claim 2, characterized in that: In step S2, the homogenization treatment may include: adding injection water to the pretreated cowhide, homogenizing the pretreated cowhide with a colloid mill, adjusting the pH value to 2.0-3.0 after homogenization, and obtaining a homogenate; wherein, the ratio of the pretreated cowhide to the injection water may be 1kg:(2-8)L; the colloid mill is pre-soaked in a 0.4-0.6M sodium hydroxide solution for not less than 30 minutes; the pH value may be adjusted to 2.0-3.0 with glacial acetic acid; the cowhide content in the homogenate may be 20-25g / L.

6. The hot spring water trace element liposome according to claim 2, characterized in that: In step S3, the adsorption filtration may include: adjusting the pH value of the homogenized digestion solution to 3.9-4.1, then adding pretreated diatomaceous earth for static adsorption, collecting the supernatant and filtering to obtain an adsorption filtrate; wherein, the preparation method of the pretreated diatomaceous earth may include: soaking the diatomaceous earth in 8-12mM hydrochloric acid solution for more than 2h, and washing it with purified water for more than 3 times after soaking to obtain the pretreated diatomaceous earth; the amount of pretreated diatomaceous earth added may be 14-16g / L of the homogenized digestion solution.

7. The hot spring water trace element liposome according to claim 2, characterized in that: In step S4, the concentration and liquid exchange and the secondary concentration and liquid exchange respectively include: using a 100kD ultrafiltration membrane, using an acetate buffer of pH 4-5 to exchange the liquid 1-3 times, and concentrating and collecting the liquid after the liquid exchange.

8. The hot spring water trace element liposome according to claim 2, characterized in that: In step S5, 5.5-6.5 mol / L hydrochloric acid solution can be used for acid hydrolysis; the amount of hydrochloric acid solution used during acid hydrolysis is 1.5-2% of the volume of the solution before hydrolysis; in addition, the acid hydrolysis temperature can be controlled to be 55-70°C, and the acid hydrolysis time can be 40-48h.

9. The hot spring water trace element liposome according to claim 2, characterized in that: The preparation method of hot spring water trace element liposomes comprises the following steps: dissolving phospholipids, ceramide and cholesterol in ether, separately taking hydrolyzed collagen, adding water to dissolve it, adding the ether solution containing phospholipids, ceramide and cholesterol into the hydrolyzed collagen aqueous solution, subjecting to short-time ultrasound or oscillation to form a stable W / O type emulsion, then removing the ether under reduced pressure evaporation conditions to obtain a gel-state substance, then dripping 0.1M pH 6.0 phosphate buffer into the above substance, vortexing and oscillating to make the gel fall off, continuing to evaporate under reduced pressure, separately taking hyaluronic acid and a preservative to add them into the solution to dissolve, and preparing hot spring water trace element liposomes.

10. Use of the hot spring water trace element liposome according to any one of claims 1 to 9 in improving skin barrier function.

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