CN112516014B

CN112516014B - Apocynum venetum extract, preparation method thereof, anti-ultraviolet, anti-oxidation and whitening cosmetic and preparation method thereof

Info

Publication number: CN112516014B
Application number: CN202110045678.1A
Authority: CN
Inventors: 单承莺; 马世宏; 陈国庆; 汪海波; 聂韡; 李淑红; 束成杰
Original assignee: Shandong Jiuxin Biotechnology Co ltd; NANJING INSTITUTE FOR COMPREHENSIVE UTILIZATION OF WILD PLANTS CHINA COOP
Current assignee: Shandong Jiuxin Biotechnology Co ltd; NANJING INSTITUTE FOR COMPREHENSIVE UTILIZATION OF WILD PLANTS CHINA COOP
Priority date: 2021-01-08
Filing date: 2021-01-08
Publication date: 2023-04-14
Anticipated expiration: 2041-01-08
Also published as: CN112516014A

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to an apocynum venetum extract and a preparation method thereof, and an anti-ultraviolet anti-oxidation whitening cosmetic and a preparation method thereof. The invention provides a apocynum venetum extract, which comprises the effective components of apocynum venetum essential oil and apocynum venetum polysaccharide. The apocynum venetum polysaccharide has excellent antioxidant and anti-ultraviolet radiation effects besides the common water-locking and moisturizing effects, and can protect the skin and isolate external stimulation, inflammation and ultraviolet pollution; the apocynum venetum essential oil has excellent tyrosinase activity inhibition effect, can inhibit the generation of melanin, and has the whitening effect. The apocynum venetum extract is applied to cosmetics, and under the combined action of the apocynum venetum extract and other components, the cosmetics have the effects of resisting ultraviolet radiation, resisting oxidation and whitening.

Description

Apocynum venetum extract, preparation method thereof, anti-ultraviolet, anti-oxidation and whitening cosmetic and preparation method thereof

Technical Field

The invention belongs to the technical field of cosmetics, and particularly relates to an apocynum venetum extract and a preparation method thereof, and an anti-ultraviolet anti-oxidation whitening cosmetic and a preparation method thereof.

Background

In areas with inflammation and strong sunshine, the sunscreen cosmetics are indispensable products for consumers. The skin is the largest organ of the human body and is the first line of defense of the human body, the parts of the human body exposed to the outside, such as the face, the neck, the upper limbs and the lower limbs, are easily damaged by ultraviolet rays, and the sun-proof cosmetics can protect the skin of the exposed part of the human body. In China, sunscreen cosmetics belong to special-purpose cosmetics, and refer to cosmetics capable of preventing or reducing skin damage caused by ultraviolet radiation.

The ultraviolet rays which cause damage to human skin are mainly divided into two wave bands of UVB (290-320 nm) and UVA (320-400 nm) according to the difference of the wavelength, wherein the UVB mainly acts on the epidermis of the skin, the UVA mainly acts on the dermis, and the depth of the UVB can reach the middle of the dermis. The ultraviolet rays are the first killer of the skin, can cause oxidative damage to the skin, can induce the generation of various Reactive Oxygen Species (ROS), and the ROS can trigger various physiological and biochemical processes, so that excessive ROS generated in cells can cause damage to biomacromolecules in the cells if not cleared in time, thereby causing the redox state of the cells to be changed, inducing apoptosis and causing acute and chronic damage to the skin.

Sunscreen cosmetics are mainly based on sunscreen agents to resist ultraviolet rays, and are divided into two main categories, namely physical sunscreen agents and chemical sunscreen agents. Physical sunscreens do not absorb ultraviolet light, and primarily function to reflect and scatter ultraviolet light, also known as inorganic sunscreens, such as titanium dioxide, zinc oxide, kaolin, talc, iron oxide, and the like; the chemical sunscreen agents mainly have the function of absorbing ultraviolet rays, and common chemical sunscreen agents comprise benzophenone, ethylhexyl salicylate and the like; however, chemical sunscreens are somewhat irritating to the skin because the molecules of the chemical sunscreens are absorbed by the skin and the process of absorbing ultraviolet light occurs inside the skin. In addition to these two traditional sunscreens, people focus on finding new bioactive substances capable of resisting ultraviolet radiation, which are safe and efficient, from animals and plants as novel sunscreens, and the research focus in the field of current sunscreen cosmetics is also hot.

Apocynum venetum L is vertical half shrub of Apocynaceae, and is mainly distributed on saline-alkali wastelands, desert edges, river banks and gobi barren beaches in provinces such as Xinjiang, qinghai, gansu and inner Mongolia in China. Besides the fibers in the stems of the apocynum venetum can be used as high-grade textile raw materials, the apocynum venetum leaves are rich in active compounds such as flavone, organic acid, polysaccharide and the like, and can be used for developing and preparing raw materials of plant-derived cosmetics. However, the apocynum venetum extract prepared by the prior art has single effective component and single skin care effect.

Disclosure of Invention

The invention provides an apocynum venetum extract which comprises apocynum venetum essential oil and apocynum venetum polysaccharide, and can effectively play roles in resisting ultraviolet radiation, resisting oxidation and whitening when being applied to cosmetics.

In order to achieve the above purpose, the invention provides the following technical scheme:

the invention provides a apocynum venetum extract, which comprises the effective components of apocynum venetum essential oil and apocynum venetum polysaccharide.

The invention also provides a preparation method of the apocynum venetum extract, which comprises the following steps:

carrying out ultrasonic extraction on the dogbane leaves to obtain a primary extract;

distilling the primary extract to obtain apocynum venetum essential oil and an extracting solution;

and filtering and extracting the extracting solution with alcohol to obtain the apocynum venetum polysaccharide.

Preferably, the power of ultrasonic extraction is 200-300W, the time is 60-120 min, and the temperature is 50-70 ℃.

Preferably, the distillation is steam distillation, and the distillation time is 2-4 h.

Preferably, the refining of the apocynum venetum polysaccharide after the apocynum venetum polysaccharide is obtained comprises enzyme hydrolysis and impurity removal.

Preferably, the impurity removal reagent is chloroform and n-butanol, and the mass ratio of the chloroform to the n-butanol is 3-6: 1.

The invention also provides the application of the apocynum venetum extract in the ultraviolet radiation resistant, antioxidant and whitening cosmetics.

The invention also provides an anti-ultraviolet antioxidant whitening cosmetic which comprises the following components in percentage by mass: 2 to 12 percent of apocynum venetum extract, 3 to 8 percent of stearic acid, 1.5 to 4 percent of octadecanol, 1 to 3 percent of isopropyl palmitate, 3 to 8 percent of vaseline, 2 to 6 percent of white oil, 3 to 8 percent of silicone oil, 0.5 to 1.5 percent of vitamin E, 1 to 3 percent of emulsifier, 2 to 5 percent of propylene glycol, 2 to 5 percent of nano titanium dioxide, 0.5 to 1.5 percent of triethanolamine, 0.3 to 1.0 percent of phenoxyethanol and the balance of water;

the apocynum venetum extract is the apocynum venetum extract in the technical scheme.

Preferably, the content of the apocynum venetum essential oil in the anti-ultraviolet, antioxidant and whitening cosmetic is 0.5-3%, and the content of the apocynum venetum polysaccharide is 1.5-9%.

The invention also provides a preparation method of the ultraviolet-resistant sunscreen whitening cosmetic, which comprises the following steps:

mixing stearic acid, octadecanol, isopropyl palmitate, vaseline, white oil, silicone oil, vitamin E and an emulsifier to obtain an oil phase system;

dispersing propylene glycol, nano titanium dioxide, apocynum venetum polysaccharide and triethanolamine in water to obtain a water phase system;

and mixing the oil phase system and the water phase system with the apocynum venetum essential oil and the phenoxyethanol to obtain the anti-ultraviolet anti-oxidation whitening cosmetic.

The invention provides an apocynum venetum extract, which comprises apocynum venetum essential oil and apocynum venetum polysaccharide. The apocynum venetum essential oil has excellent tyrosinase activity inhibition effect, can inhibit the generation of melanin, and has the whitening effect; the apocynum venetum polysaccharide has excellent antioxidant and anti-ultraviolet radiation effects besides the common water-locking and moisturizing effects, and can protect the skin and isolate external irritation, inflammation and ultraviolet pollution. The apocynum venetum essential oil and the apocynum venetum polysaccharide are applied to the cosmetics, and under the combined action of the apocynum venetum essential oil and other components, the cosmetics have the effects of ultraviolet radiation resistance, oxidation resistance and whitening.

Drawings

FIG. 1 shows the result of the scavenging activity of Apocynum venetum polysaccharide on superoxide anion free radicals.

FIG. 2 shows the result of hydroxyl radical scavenging activity test by Apocynum venetum polysaccharide.

FIG. 3 shows the result of the anti-lipid peroxidation activity test of Apocynum venetum polysaccharides.

FIG. 4 shows the result of tyrosinase activity inhibition test by apocynum venetum essential oil.

Detailed Description

In the present invention, as the raw material, commercially available products known to those skilled in the art may be used unless otherwise specified.

The invention also provides a preparation method of the apocynum venetum extract in the technical scheme, which comprises the following steps:

The invention carries out ultrasonic extraction on dogbane leaves to obtain a primary extract. In the invention, the mode of ultrasonic extraction is preferably that the dogbane leaves are crushed and then added into water for ultrasonic extraction; the mass ratio of the dogbane leaves to the water is 1: 10-30, the further optimization is 1: 12-28, and the further optimization is 1: 15-25. In the invention, the power of the ultrasonic extraction is preferably 200-300W, more preferably 210-290W, and even more preferably 220-280W; in an embodiment of the invention, the power of the ultrasound extraction is specifically 250W. In the invention, the time of ultrasonic extraction is preferably 60-120 min, more preferably 70-110 min, and even more preferably 80-100 min; in the embodiment of the present invention, the time of ultrasonic extraction is specifically 90min. In the present invention, the temperature of the ultrasonic extraction is preferably 50 to 70 ℃, more preferably 52 to 68 ℃, and still more preferably 55 to 65 ℃.

After the primary extract is obtained, the invention distills the primary extract to obtain the apocynum venetum essential oil and the extracting solution. Steam distillation is preferably employed in the present invention. In the present invention, the distillation time is preferably 2 to 4 hours, more preferably 2.2 to 3.8 hours, and still more preferably 2.5 to 3.5 hours. In the invention, the volatile oil collected after distillation is the apocynum venetum essential oil, and the residual base solution is the extracting solution.

After the extracting solution is obtained, the extracting solution is filtered and extracted by alcohol to obtain the apocynum venetum polysaccharide. In the present invention, it is preferable that the extract is filtered while it is hot to obtain a supernatant.

Alcohol is carried out in advance, the supernatant is preferably concentrated by the method of concentrating the supernatant by reduced pressure distillation, and the volume of the concentrated supernatant is preferably 1/5 of that of the supernatant. The invention has no special requirements on the condition parameters of the reduced pressure distillation concentration, and can obtain the concentrated solution with the required volume. In the invention, the alcohol extraction mode is preferably to add 95% ethanol into the concentrated solution, and the concentrated solution is stood and then centrifuged to obtain sediment, namely the apocynum venetum polysaccharide; the volume ratio of the ethanol to the concentrated solution is preferably 2-5: 1, more preferably 2.5-4.5: 1, and even more preferably 3-4: 1; the standing time is preferably 20 to 30 hours, more preferably 22 to 28 hours, and still more preferably 24 hours. According to the invention, the apocynum venetum essential oil and polysaccharide components can be simultaneously obtained through one-time heating water extraction process by adopting ultrasonic-assisted steam distillation-hot water coupling extraction, so that on one hand, the yield of the essential oil and the polysaccharide can be improved, and on the other hand, the preparation method is simple, convenient, efficient and energy-saving.

After the apocynum venetum polysaccharide is obtained, the apocynum venetum polysaccharide is preferably refined in a refining mode, wherein the refining mode preferably comprises enzyme hydrolysis and impurity removal. In the invention, the enzymatic hydrolysis is preferably carried out by adding papain into an apocynum venetum polysaccharide solution, and the mass concentration of the apocynum venetum polysaccharide solution is preferably 5-10%, more preferably 6-9%, and even more preferably 7-8%; the papain accounts for 2-4 wt.%, preferably 2.5-3.5 wt.%, and more preferably 3wt.% of the apocynum venetum polysaccharide; the enzymatic hydrolysis is preferably carried out under heated conditions; the heating mode is preferably water bath heating, and the heating temperature is preferably 40-50 ℃, more preferably 42-48 ℃, and more preferably 45 ℃; the heating time is preferably 1 to 3 hours, more preferably 1.5 to 2.5 hours, and still more preferably 2 hours.

After the enzymatic hydrolysis, the method preferably removes impurities from the hydrolyzed solution, and the method preferably comprises the steps of adding an impurity removal reagent into the solution for extraction to obtain a water phase part; in the embodiment of the invention, the impurity removal reagent is specifically chloroform and n-butanol, and the mass ratio of the chloroform to the n-butanol is preferably 3-6: 1, more preferably 3.5-5.5: 1, and more preferably 4-5: 1; the dosage of the impurity removal reagent is preferably 1/10-1/2, more preferably 1/5-2/5, and even more preferably 1/3 of the volume of the polysaccharide solution. After the water phase part is obtained, the invention preferably carries out concentration and alcohol extraction on the water phase part to obtain the apocynum venetum polysaccharide; the concentration is preferably carried out by distillation under reduced pressure, and the volume after concentration is preferably 1/5 of the volume of the aqueous phase part. The invention has no special requirements on the condition parameters of the reduced pressure distillation concentration, and can obtain the concentrated solution with the required volume. After the concentrated solution is obtained, the concentrated solution is preferably subjected to alcohol extraction, the alcohol extraction mode is preferably to add 95% ethanol into the concentrated solution, the concentrated solution is kept stand and then centrifuged, and the obtained precipitate is the apocynum venetum polysaccharide; the volume ratio of the ethanol to the concentrated solution is preferably 2-5: 1, more preferably 2.5-4.5: 1, and even more preferably 3-4: 1; the standing time is preferably 20 to 30 hours, more preferably 22 to 28 hours, and still more preferably 24 hours. The apocynum venetum polysaccharide obtained after alcohol extraction is preferably dried, and the drying mode is preferably vacuum freeze drying; the invention has no special requirements on the parameters of vacuum drying, and can obtain the dried apocynum venetum polysaccharide. The invention adopts the modes of enzymatic hydrolysis and impurity removal for refining, can effectively remove protein in the polysaccharide, and ensures that the purity of the polysaccharide is higher.

The invention also provides an anti-ultraviolet antioxidant whitening cosmetic which comprises the following components in percentage by mass: 2 to 12 percent of apocynum venetum extract, 3 to 8 percent of stearic acid, 1.5 to 4 percent of octadecanol, 1 to 3 percent of isopropyl palmitate, 3 to 8 percent of vaseline, 2 to 6 percent of white oil, 3 to 8 percent of silicone oil, 0.5 to 1.5 percent of vitamin E, 1 to 3 percent of emulsifier, 2 to 5 percent of propylene glycol, 2 to 5 percent of nano titanium dioxide, 0.5 to 1.5 percent of triethanolamine, 0.3 to 1.0 percent of phenoxyethanol and the balance of water; the apocynum venetum extract is apocynum venetum essential oil and apocynum venetum polysaccharide which are prepared by the technical scheme.

The anti-ultraviolet antioxidant whitening cosmetic provided by the invention comprises 2-12% of apocynum venetum extract, more preferably 3-11%, more preferably 4-10% by mass, wherein the content of the apocynum venetum essential oil in the anti-ultraviolet antioxidant whitening cosmetic is 0.5-3%, more preferably 1-2.5%, more preferably 1.5-2%; the content of the apocynum venetum polysaccharide is 1.5 to 9 percent, more preferably 2 to 8.5 percent, and still more preferably 2.5 to 8 percent. In the invention, the apocynum venetum polysaccharide has good ultraviolet radiation resistance and oxidation resistance besides the effects of water locking and moisture retention; the apocynum venetum essential oil can inhibit the activity of tyrosinase and the generation of melanin, and has the whitening effect.

The anti-ultraviolet antioxidant whitening cosmetic provided by the invention comprises 3-8% of stearic acid by mass percentage, more preferably 3.5-7.5% of stearic acid by mass percentage, and even more preferably 4-7% of stearic acid by mass percentage. In the invention, the stearic acid can play a role in lubrication and emulsification, and can change the cosmetics into a stable white paste.

The anti-ultraviolet, antioxidant and whitening cosmetic provided by the invention comprises 1.5-4% of octadecanol, more preferably 2-3.5%, and even more preferably 2.5-3% by mass. In the invention, the octadecanol is a skin conditioner and an emulsion stabilizer.

The ultraviolet-resistant, antioxidant and whitening cosmetic provided by the invention comprises 1-3% of isopropyl palmitate, more preferably 1.2-2.8%, and even more preferably 1.5-2.5% by mass. In the invention, the isopropyl palmitate can increase the ductility of the cosmetic, so that the cosmetic is fine, smooth, bright and non-greasy.

The uvioresistant, antioxidant and whitening cosmetic provided by the invention comprises 3-8% of vaseline, more preferably 3.5-7.5%, and even more preferably 4-7% by mass. In the invention, the vaseline can play a role in moisturizing and nourishing the skin.

The anti-ultraviolet antioxidant whitening cosmetic provided by the invention comprises 2-6% of white oil by mass percentage, more preferably 2.5-5.5% of white oil by mass percentage, and more preferably 3-5% of white oil by mass percentage. In the invention, the white oil can block the moisture evaporation of the skin and plays a role in lubrication and moisture preservation in cosmetics.

The anti-ultraviolet antioxidant whitening cosmetic provided by the invention comprises 3-8% of silicone oil by mass percentage, more preferably 3.5-7.5% of silicone oil by mass percentage, and more preferably 4-7% of silicone oil by mass percentage. In the invention, the silicone oil can play a moisturizing and lubricating role in the cosmetics, can regulate skin feel and enables the cosmetics to be more easily pushed away.

The ultraviolet-resistant, oxidation-resistant and whitening cosmetic provided by the invention comprises 0.5-1.5% of vitamin E by mass, more preferably 0.6-1.4% of vitamin E by mass, and more preferably 0.7-1.3% of vitamin E by mass. In the present invention, the vitamin E can accelerate skin metabolism, promote skin repair and regeneration.

The ultraviolet-resistant, oxidation-resistant and whitening cosmetic provided by the invention comprises 1-3% of emulsifier by mass, more preferably 1.2-2.8% of emulsifier by mass, and more preferably 1.5-2.5% of emulsifier by mass. In the invention, the emulsifier can emulsify the essential oil components in the cosmetic, so that the essential oil is more stable in the cosmetic.

The anti-ultraviolet antioxidant whitening cosmetic provided by the invention comprises 2-5% of propylene glycol, more preferably 2.5-4.5%, and even more preferably 3-4% by mass. In the present invention, the propylene glycol is capable of keeping the skin moist.

The ultraviolet-resistant, oxidation-resistant and whitening cosmetic provided by the invention comprises 2-5% of nano titanium dioxide, more preferably 2.5-4.5%, and more preferably 3-4% by mass. In the invention, the nano titanium dioxide is used as a physical sun-screening agent to absorb and obstruct ultraviolet rays.

The anti-ultraviolet oxidation-resistant whitening cosmetic provided by the invention comprises, by mass, 0.5-1.5% of triethanolamine, more preferably 0.6-1.4%, and even more preferably 0.7-1.3%. In the invention, the triethanolamine is used as an emulsifier and a humectant, so that the cream of the cosmetic is fine and smooth and is bright and white.

The anti-ultraviolet antioxidant whitening cosmetic provided by the invention comprises 0.3-1.0% of phenoxyethanol, more preferably 0.4-0.9%, and even more preferably 0.5-0.8% by mass. In the invention, the phenoxyethanol can play a role of a preservative.

The invention also provides a preparation method of the ultraviolet-resistant, antioxidant and whitening cosmetic, which comprises the following steps:

The oil phase system is obtained by mixing stearic acid, octadecanol, isopropyl palmitate, vaseline, white oil, silicone oil, vitamin E and an emulsifier.

Preferably, stearic acid, octadecanol, isopropyl palmitate and vaseline are mixed and stirred uniformly under the heating condition; in the present invention, the heating temperature is preferably 60 to 75 ℃, more preferably 62 to 73 ℃, and still more preferably 65 to 70 ℃; the invention has no special requirements on the stirring condition and can be mixed uniformly. After uniformly mixing, preferably adding white oil, silicone oil, vitamin E and an emulsifier, and uniformly stirring under a heating condition; in the present invention, the heating temperature is preferably 85 to 95 ℃, more preferably 87 to 93 ℃, and still more preferably 85 to 90 ℃, and the stirring time is preferably 15 to 30min, more preferably 18 to 27min, and still more preferably 20 to 25min. After stirring uniformly, the mixture is preferably cooled to 65-75 ℃ to obtain an oil phase system. Mixing at the above temperatures can make the components in the oil phase system more miscible with each other.

The invention disperses propanediol, nanometer titanium pigment, apocynum venetum polysaccharide and triethanolamine in water to obtain a water phase system.

The invention preferably disperses propylene glycol, nano titanium dioxide and apocynum venetum polysaccharide in water, and the mixture is stirred uniformly under the heating condition; in the present invention, the heating temperature is preferably 45 to 60 ℃, more preferably 47 to 58 ℃, and still more preferably 45 to 55 ℃; after stirring uniformly, preferably heating to 65-75 ℃ under stirring, and adding triethanolamine to obtain an aqueous phase system. The emulsifying effect of the aqueous system can be improved at the above temperature.

The anti-ultraviolet antioxidant whitening cosmetic is prepared by mixing the oil phase system, the water phase system, the apocynum venetum essential oil and phenoxyethanol. The invention preferably mixes the oil phase system and the water phase system at 65-75 ℃, evenly stirs, adds the apocynum venetum essential oil after the temperature of the system is reduced to 50-60 ℃, evenly stirs, and adds the phenoxyethanol to obtain the anti-ultraviolet anti-oxidation whitening cosmetic. In the invention, the preparation process can prevent the volatilization of the essential oil and can ensure that all components are better emulsified in the product.

In order to further illustrate the present invention, the apocynum venetum extract and the ultraviolet-resistant, antioxidant and whitening cosmetic provided by the present invention will be described in detail below with reference to the accompanying drawings and examples, which should not be construed as limiting the scope of the present invention.

Example 1

Pulverizing folium Apocyni Veneti, adding deionized water at a mass ratio of 1: 20, and performing ultrasonic treatment at 60 deg.C and 250W for 90min to obtain primary extract.

And (3) carrying out steam distillation on the primary extract for 4 hours, wherein the collected volatile oil is the apocynum venetum essential oil, the residual base solution is the extracting solution, and the extraction rate of the apocynum venetum essential oil is 0.12-0.14%.

Filtering the hot extract, distilling the supernatant under reduced pressure, concentrating to 1/5 of the volume of the supernatant to obtain a concentrated solution, adding 95% ethanol with the volume 4 times that of the concentrated solution, standing for 24h, and centrifuging to obtain a precipitate, namely the apocynum venetum polysaccharide.

Preparing apocynum venetum polysaccharide into a solution with the mass concentration of 6%, and adding 3wt.% of papain; heating in water bath at 45 deg.C for 2 hr, adding impurity removing reagent (mass ratio of chloroform to n-butanol is 5: 1) with volume of 1/3 of polysaccharide solution, and extracting to obtain water phase. Distilling and concentrating the water phase part to 1/5 of the volume, adding 95% ethanol with the volume 4 times that of the concentrated solution, standing for 24h, centrifuging, and performing vacuum freeze drying on the precipitate to obtain the apocynum venetum polysaccharide.

Example 2

The anti-ultraviolet antioxidant whitening cosmetic prepared from the apocynum venetum polysaccharide obtained in the example 1 and the apocynum venetum essential oil is prepared according to the following formula components: 5% of stearic acid, 2.5% of octadecanol, 1.5% of isopropyl palmitate, 5% of vaseline, 4% of white oil, 5% of silicone oil, 1% of vitamin E, 2% of apocynum venetum essential oil, 2% of emulsifier, 3% of propylene glycol, 5% of apocynum venetum polysaccharide, 3% of nano titanium dioxide, 1% of triethanolamine, 0.5% of phenoxyethanol and the balance of deionized water.

Mixing stearic acid, octadecanol, isopropyl palmitate and vaseline, and stirring uniformly at 65 ℃; adding white oil, silicone oil, vitamin E and emulsifier, stirring at 90 deg.C for 25min, and cooling to 70 deg.C to obtain oil phase system; dispersing propylene glycol, nano titanium dioxide and apocynum venetum polysaccharide in water, and uniformly stirring at 50 ℃; heating to 70 deg.C, adding triethanolamine to obtain water phase system.

Mixing the oil phase system and the water phase system at 70 deg.C, cooling to 55 deg.C, adding herba Apocyni Veneti essential oil, stirring, and adding phenoxyethanol to obtain the final product.

Example 3

Preparing the anti-ultraviolet, anti-oxidation and whitening cosmetic from the apocynum venetum polysaccharide obtained in the example 1 and the apocynum venetum essential oil according to the following formula components: 3% of stearic acid, 4% of octadecanol, 1% of isopropyl palmitate, 8% of vaseline, 2% of white oil, 8% of silicone oil, 0.5% of vitamin E, 3% of apocynum venetum essential oil, 3% of emulsifier, 5% of propylene glycol, 2% of apocynum venetum polysaccharide, 5% of nano titanium dioxide, 0.5% of triethanolamine, 0.3% of phenoxyethanol and the balance of deionized water.

Mixing stearic acid, octadecanol, isopropyl palmitate and vaseline, and stirring uniformly at 60 ℃; adding white oil, silicone oil, vitamin E and emulsifier, stirring at 85 deg.C for 30min, and cooling to 65 deg.C to obtain oil phase system; dispersing propylene glycol, nano titanium dioxide and apocynum venetum polysaccharide in water, and uniformly stirring at 45 ℃; heating to 65 ℃, and adding triethanolamine to obtain a water phase system.

Mixing the oil phase system and the water phase system at 65 deg.C, cooling to 50 deg.C, adding herba Apocyni Veneti essential oil, stirring, and adding phenoxyethanol to obtain the final product.

Example 4

Preparing the anti-ultraviolet, anti-oxidation and whitening cosmetic from the apocynum venetum polysaccharide obtained in the example 1 and the apocynum venetum essential oil according to the following formula components: 8% of stearic acid, 1.5% of octadecanol, 3% of isopropyl palmitate, 3% of vaseline, 6% of white oil, 3% of silicone oil, 1.5% of vitamin E, 0.5% of apocynum venetum essential oil, 1% of emulsifier, 2% of propylene glycol, 8% of apocynum venetum polysaccharide, 2% of nano titanium dioxide, 1.5% of triethanolamine, 1.0% of phenoxyethanol and the balance of deionized water.

Mixing stearic acid, octadecanol, isopropyl palmitate and vaseline, and stirring uniformly at 75 ℃; adding white oil, silicone oil, vitamin E and emulsifier, stirring at 95 deg.C for 15min, and cooling to 75 deg.C to obtain oil phase system; dispersing propylene glycol, nano titanium dioxide and apocynum venetum polysaccharide in water, and uniformly stirring at 60 ℃; heating to 75 deg.C, adding triethanolamine to obtain water phase system.

Mixing the oil phase system and the water phase system at 75 deg.C, cooling to 60 deg.C, adding herba Apocyni Veneti essential oil, stirring, and adding phenoxyethanol to obtain the final product.

And (3) verifying the antioxidant performance of the apocynum venetum polysaccharide:

example 5

The apocynum venetum polysaccharide obtained in example 1 was used for the determination of scavenging activity of superoxide anion radicals.

Preparing apocynum venetum polysaccharide solutions with mass concentrations of 0.2, 0.4, 0.6, 0.8 and 1.0mg/mL, precisely measuring 6.50mL of 0.05mol/L Tris-HCl buffer solution (pH 8.2), placing in a water bath at 25 ℃, preheating for 20min, respectively adding 1.00mL of apocynum venetum polysaccharide solutions with different concentrations and 0.40mL of 25mmol/L pyrogallol solution, mixing uniformly, placing in the water bath at 25 ℃ for reaction for 5min, and adding 1.00mL of 8mmol/L HCl solution to terminate the reaction. The absorbance was measured at a wavelength of 325 nm. Three replicates of each sample were taken and the average value was A3. And the positive control is vitamin C, corresponding background absorbance values A1, A2 and A4 are respectively determined by referring to the operations, and the superoxide anion clearance rate is calculated according to the following formula:

clearance = { [ (A1-A2) - (A3-A4) ]/(A1-A2) } 100%

A1: absorbance value without sample;

a2: absorbance values without sample and pyrogallol;

a3: (ii) an absorbance value comprising the sample and pyrogallol;

a4: absorbance values with sample, but no pyrogallol.

The results of the test for scavenging activity of apocynum venetum polysaccharide on superoxide anion radicals are shown in table 1 and fig. 1.

TABLE 1 Apocynum venetum polysaccharide scavenging activity test results for superoxide anion radical

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As can be seen from Table 1, in the experimental concentration range, with the increase of the concentrations of vitamin C and apocynum venetum polysaccharide, the clearance rates of vitamin C and apocynum venetum polysaccharide to superoxide anion free radicals are also obviously increased, and the median Inhibitory Concentration (IC) of the apocynum venetum polysaccharide is also obviously increased ₅₀ ) A value of about 0.38mg/mL, slightly above the median inhibitory concentration of vitamin C (0.28 mg/mL); this indicates that the scavenging activity of the apocynum venetum polysaccharide and the vitamin C on superoxide anion free radicals is in the same order of magnitude.

Example 6

The apocynum venetum polysaccharide obtained in example 1 was subjected to an activity measurement experiment for scavenging hydroxyl radicals.

Preparing apocynum venetum polysaccharide solution with mass concentration of 0.2, 0.4, 0.6, 0.8 and 1.0mg/mL, and adopting improved Smimoff salicylic acid method, namely using H ₂ O ₂ With Fe ²⁺ The reaction generates hydroxyl radical, salicylic acid is added into the system to capture the hydroxyl radical and generate a colored substance, and the substance has maximum absorption at 510 nm. 10mmol/L FeSO per tube is added into a 10mL test tube with a plug ₄ 1mL of each solution and 10mmol/L salicylic acid solution and 1mL of apocynum venetum polysaccharide solution with different concentrations, and finally adding 9mmol/L H ₂ O ₂ Starting the reaction with 1mL of the solution, adding distilled water to 10mL, reacting in a water bath at 37 ℃ for 30min, then zeroing with distilled water, and determining 51Absorbance at 0 nm. Three replicates of each sample were taken and averaged for A1. And the positive control is vitamin C, and the corresponding background absorbance values A2 are respectively determined by referring to the operations, and the hydroxyl radical clearance rate is calculated according to the following formula:

clearance = [1- (A1-A2)/A0 ] × 100%

A0: adding the sample into the system instead of 1mL of distilled water without adding the sample;

a1: sample adding reaction;

a2: adding sample but not reacting, i.e. replacing H with 1mL of distilled water ₂ O ₂ The solution was added to the system.

The results of the test for the scavenging activity of apocynum venetum polysaccharide on the super hydroxyl radical are shown in table 2 and fig. 2.

TABLE 2 Apocynum venetum polysaccharide for scavenging activity of hydroxyl radical test results

As can be seen from Table 2, with the increase of the concentration, the effect of eliminating hydroxyl free radicals of the apocynum venetum polysaccharide is obvious, and the elimination capability and the concentration have obvious dose-effect relationship, namely IC ₅₀ The value is almost equal to that of the vitamin C, which indicates that the apocynum venetum polysaccharide obtained by the invention has the capability of eliminating hydroxyl free radicals equivalent to that of the vitamin C.

Example 7

The apocynum venetum polysaccharide obtained in example 1 is used for the determination experiment of the anti-lipid peroxidation activity.

Preparing apocynum venetum polysaccharide solutions with mass concentrations of 0.2, 0.4, 0.6, 0.8 and 1.0mg/mL, precisely measuring 0.50mL of lecithin solution, adding 1.00mL of PBS buffer solution with the pH value of 7.4, 1.00mL of apocynum venetum polysaccharide solution with different concentrations and 1.00mL of 2.5mmol/L EDTA-Fe (II), uniformly mixing, reacting in a water bath at 37 ℃ for 45min, adding 2.00mL of trichloroacetic acid with 28% (w/v), 1.00mL of thiobarbituric acid with 1% (w/v), uniformly mixing, placing in a boiling water bath at 100 ℃ for heating for 10min, cooling, and measuring the absorbance at a position of 532 nm. The absorbance A0 was measured by zeroing with PBS buffer and replacing the sample with PBS buffer for blank tubes. Three replicates of each sample were taken and averaged for A1. The positive control is vitamin C. The lipid peroxidation inhibition was calculated as follows:

inhibition (%) = [ (A0-A1)/A0 ]. 100%

A0: absorbance of no sample;

a1: including the absorbance of the sample.

The results of the anti-lipid peroxidation activity test of the apocynum venetum polysaccharides are shown in table 3 and fig. 3.

TABLE 3 Apocynum venetum polysaccharide anti-lipid peroxidation activity test results

As can be seen from Table 3, apocynum venetum polysaccharide vs. Fe ²⁺ The induced peroxidation of the lecithin liposome has an inhibiting effect, and the inhibition rate is slowly increased along with the increase of the concentration and is equivalent to the inhibition rate of vitamin C on the lipid peroxidation.

Superoxide anion free radicals are harmful free radicals closely related to the human body, attack biological macromolecules, cause the crosslinking or the breakage of the biological macromolecules, cause the destruction of cell structures and functions and cause aging; hydroxyl free radical is the most active free radical in human body and has the greatest harm to organism, and can react with lipid, polypeptide, protein, DNA and the like to initiate the oxidation of unsaturated fatty acid to form Lipid Peroxide (LPO) to destroy membrane structure, thus causing damage to cells, tissues and organs, inducing various diseases and accelerating the aging of organism; lipid peroxidation can damage biological membranes and their functions, leading to cell transparency lesion, fibrosis, and extensive cell damage leading to damage to skin, nerves, tissues, organs, etc. As shown in tables 1-3, the apocynum venetum polysaccharide has good capacity of removing superoxide anion free radicals and hydroxyl free radicals, has good inhibition effect on lipid peroxidation, and has excellent in-vitro antioxidant activity.

And (3) verifying the ultraviolet radiation resistance of the apocynum venetum polysaccharide:

example 8

The apocynum venetum polysaccharide obtained in example 1 is subjected to an anti-ultraviolet radiation test, and an in vitro radiation test is performed by using escherichia coli as a model thallus to detect the anti-ultraviolet radiation protection effect of the apocynum venetum polysaccharide.

1. Preparation of suspension liquid

The frozen seed was removed and prepared in sterile test tubes (with stoppered plugs), sterilized LB broth and pipette. In a clean bench, firstly adding 1/2 LB liquid culture medium into a test tube with a plug, then adding 0.5mL of strain into the test tube by using a liquid transfer gun, finally covering the test tube with the plug, putting the test tube into a shaking table for culturing for 18h-24h at 37 ℃ and 180rpm, and taking out the test tube and refrigerating the test tube for later use.

2. Preparation of polysaccharide solutions of different concentrations

Accurately weighing 5g of apocynum venetum polysaccharide powder, dissolving the apocynum venetum polysaccharide powder in 100mL of sterile physiological saline to prepare a solution with the concentration of 5%, diluting ten times to obtain a 0.5% solution, and refrigerating the solution for later use.

3. Preparation of solid nutrient agar medium

Accurately weighing 10g of tryptone, 5g of yeast extract, 10g of sodium chloride and 20g of agar powder, putting the weighed materials into a beaker, adding 1000mL of distilled water into the beaker, heating and stirring the materials to fully dissolve the materials, pouring the materials into a conical flask for sealing, putting the conical flask into an autoclave for sterilization at 121 ℃ for 20min, subpackaging the materials after the sterilization is finished, pouring the materials into a sterilization flat plate, and putting the sterilized flat plate into a refrigerator for cold storage after complete cooling and solidification for later use.

4. In vitro thallus radiation experiment

Taking three sterilized plates, adding 1mL of the previously prepared bacterial suspension into each plate by using a pipette, adding 9mL of apocynum venetum polysaccharide solutions with different concentrations into two plates respectively, and adding 9mL of sterile physiological saline into the two plates as a blank control group. After mixing the mixture sufficiently, the flat plate is placed under an ultraviolet lamp, and the timing is started from the turning on of the ultraviolet lamp and is stopped when the ultraviolet lamp is turned off. Respectively taking out 100 mu L of mixed bacteria liquid from the flat plates which are irradiated by ultraviolet light for different time, gradually diluting the mixed bacteria liquid by using sterile normal saline according to different concentration requirements, and finally taking 20 mu L of the diluted mixed bacteria liquid by using a pipette and respectively adding the 20 mu L of the diluted mixed bacteria liquid into the nutrient agar culture medium flat plates which are correspondingly marked. And finally, putting the culture medium plate added with the mixed bacterial liquid into an incubator to be cultured for 24 hours at 37 ℃.

Through observation results, the optimal dilution factor of the bacterial suspension is determined to be 10 ³ The ultraviolet irradiation time is 0min, 5min, 10min and 15min, and the change of the number of the viable bacteria is observed under the condition. The experimental parameter design is shown in table 4, and the experimental results are shown in table 5.

TABLE 4 anti-ultraviolet radiation experimental design of mixed bacteria liquid of Escherichia coli

Serial number of culture dish	Dilution factor	Irradiation time/min	Adding sugar solution to obtain sugar solution with high concentration	Sterile normal saline
					1		0		√
2		5		√
					3		10		√
4		15		√
					5	10 ³	0	0.5％
6	10 ³	5	0.5％
					7	10 ³	10	0.5％
8	10 ³	15	0.5％
					9	10 ³	0	5％
10	10 ³	5	5％
					11	10 ³	10	5％
12	10 ³	15	5％

TABLE 5 ultraviolet radiation resistance test results of Escherichia coli mixed bacteria liquid

Note: the number of colonies represented by ≥ 20, + represents ≥ 10, + represents ≥ 5, -represents the number of colonies visible without the naked eye.

As is apparent from Table 5, only a few surviving E.coli cells remained in the blank control group after 5min of UV irradiation, and no surviving E.coli cells remained on the medium after 10min. In the apocynum venetum polysaccharide solution group with the concentration of 0.5%, the survival number of escherichia coli is gradually reduced from the culture medium in three time periods of 5min, 10min and 15min, but the escherichia coli still survives on the culture medium after the irradiation for 15 min. In the case of the apocynum venetum polysaccharide solution group having a concentration of 5%, the number of Escherichia coli cells was gradually decreased in the medium in three periods of 5min, 10min and 15min, but a large number of viable Escherichia coli cells were present in the medium even after 15min of irradiation. In conclusion, the apocynum venetum polysaccharide solution is irradiated by ultraviolet rays under the same environment and time, the protection effect on the ultraviolet radiation resistance of escherichia coli is presented, and the effect is increased along with the increase of the concentration.

And (3) verifying the performance of the apocynum venetum essential oil for inhibiting tyrosinase:

example 9

The apocynum venetum essential oil obtained in example 1 was subjected to tyrosinase inhibition test.

1. Reagent preparation

Sodium phosphate buffer (1/15m, ph = 6.8): 1.0001g of sodium dihydrogen phosphate and 1.1860g of disodium hydrogen phosphate are accurately weighed, a small amount of deionized water is added for dissolution, the volume is determined to 500mL, and the mixture is stored in a refrigerator at 4 ℃ for standby.

L-tyrosine solution (7.5 mmol/L): accurately weighing 0.2721g of L-tyrosine, adding a plurality of drops of concentrated hydrochloric acid, adding about 50mL of deionized water, adjusting the pH to 7.0 by using a sodium hydroxide solution after the solution is completely dissolved by slight heating, and adding deionized water to a constant volume of 200mL.

Test solution: accurately weighing 0.1000g of apocynum venetum essential oil, dissolving the apocynum venetum essential oil in 20mL of dimethyl sulfoxide to obtain 5mg/mL of solution to be detected, and diluting the solution to 2.5mg/mL, 1.25mg/mL, 0.625mg/mL, 0.3125mg/mL and 0.15625mg/mL in a double way.

Positive control (+ CK): accurately weighing 0.1000g arbutin powder, dissolving in 20mL deionized water to obtain 5.00mg/mL positive control mother liquor, and diluting to 2.5mg/mL, 1.2500mg/mL, 0.6250mg/mL, 0.3125mg/mL, 0.1562mg/mL.

2. Preparation of tyrosinase liquid

Cleaning potato, and precooling at 4 deg.C for about 4h. Peeling, cutting into pieces of about 1X 1cm3, and freezing at-20 deg.C overnight. Adding 4 deg.C pre-cooled sodium phosphate buffer solution at a ratio of 1: 1 (W: V), homogenizing with tissue triturator, filtering with 3 layers of gauze, centrifuging the filtrate at 4000r/min for 10min to obtain supernatant as obtained tyrosinase crude enzyme solution, storing at 4 deg.C, and using up within 2h.

3. Inhibition of tyrosinase activity by samples

Adding phosphate buffer solution, test solution with different concentration gradients (including positive control), and enzyme solution into the test tube, and water-bathing at 30 deg.C for 10min. The substrate L-tyrosine was then added and the timer was started immediately. The absorbance at a wavelength of 475nm was measured at 20min of the reaction. When in measurement, the inhibition rate of the test solution (including a positive control) on tyrosinase is calculated by using a corresponding negative control as a reference through the following formula, and the half Inhibition Concentration (IC) is estimated according to a concentration-enzyme inhibition rate curve ₅₀ ) An approximation of (d). Each experiment was performed in 3 replicates. The experimental parameter settings of the inhibition activity of tyrosinase by apocynum venetum essential oil are shown in table 6, and the test results are shown in table 7 and fig. 4.

Inhibition = [ (A-B)/A ]. 100%

Wherein "A" is the absorbance of the standard control and "B" is the absorbance of the test solution (or positive control).

TABLE 6 test parameter design for inhibition of tyrosinase activity by apocynum venetum essential oil

TABLE 7 Apocynum venetum essential oil tyrosinase inhibitory activity test results

As can be seen from Table 7, in the experimental concentration range, the inhibition rate of tyrosinase by the sample and the positive control showed an increasing trend along with the increase of the concentration, and the inhibition rate showed dose-dependent inhibition. In the experimental concentration range, the inhibition rate of the apocynum venetum essential oil on tyrosinase is obviously higher than that of arbutin.

Example 10

The ultraviolet-resistant, antioxidant and whitening cosmetic obtained in example 2 is used for volunteer trial tests.

The test population: 20 volunteers, all women of 18-55 years old, were healthy for 30 days.

The sunscreen cosmetic is applied to the exposed skin 15min before the person comes out, and if the period is more than 3h outdoors, the sunscreen cosmetic needs to be applied again. The results of the trial are shown in Table 8.

TABLE 8 trial results of ultraviolet-resistant, antioxidant, and whitening cosmetics

As can be seen from the table 8, after the trial for 30 days, all the tested people do not find red itch and irritation reaction, and the self-describing feeling is smooth and comfortable without greasy feeling, so that the electric heating water is particularly suitable for being used in summer.

As can be seen from the above examples, the apocynum venetum polysaccharide has excellent in vitro oxidation resistance and ultraviolet radiation resistance, and the apocynum venetum essential oil can inhibit the activity of tyrosinase. The cosmetic added with the apocynum venetum polysaccharide and the apocynum venetum essential oil can control the generation of melanin, has the whitening effect, can actively protect the skin of a human body, isolate the skin aging problem and the like caused by external stimulation, inflammation, ultraviolet pollution and the like, and can keep the skin in a relatively durable moist and healthy state.

Although the above embodiments have been described in detail, they are only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and all of the embodiments belong to the protection scope of the present invention.

Claims

1. An apocynum venetum extract is used as the only active component in the preparation of cosmetics with the effects of resisting ultraviolet radiation, resisting oxidation and whitening;

the active ingredients of the apocynum venetum extract are apocynum venetum essential oil and apocynum venetum polysaccharide;

the content of the apocynum venetum essential oil in the cosmetic with the ultraviolet radiation resisting, oxidation resisting and whitening effects is 0.5 to 3 percent, and the content of the apocynum venetum polysaccharide is 1.5 to 9 percent;

the preparation method of the apocynum venetum extract comprises the following steps:

pulverizing folium Apocyni Veneti, adding deionized water at a mass ratio of 1: 20, and performing ultrasonic treatment at 60 deg.C and 250W for 90min to obtain primary extract;

performing steam distillation on the primary extract for 4h, wherein the collected volatile oil is apocynum venetum essential oil, and the residual base solution is an extracting solution;

filtering the extracting solution while the extracting solution is hot, taking the supernatant, carrying out reduced pressure distillation and concentration on the supernatant until the volume of the supernatant is 1/5 of that of the supernatant to obtain a concentrated solution, adding 95% ethanol with the volume being 4 times that of the concentrated solution, standing for 24 hours, and centrifuging to obtain a precipitate, namely the apocynum venetum polysaccharide;

preparing the apocynum venetum polysaccharide into a solution with the mass concentration of 6%, and adding 3wt.% of papain; heating in water bath at 45 deg.C for 2 hr, adding impurity removing reagent with volume of 1/3 of polysaccharide solution, and extracting to obtain water phase; the impurity removal reagent comprises chloroform and n-butanol; the mass ratio of the chloroform to the n-butanol is 5: 1;

and (3) distilling and concentrating the water phase part to 1/5 of the volume, adding 95% ethanol with the volume being 4 times that of the concentrated solution, standing for 24 hours, centrifuging, and carrying out vacuum freeze-drying on the precipitate to obtain the apocynum venetum polysaccharide.

2. The application of the cosmetic with the ultraviolet radiation resisting, oxidation resisting and whitening effects as claimed in claim 1 is characterized by comprising the following components in percentage by mass: 5% of stearic acid, 2.5% of octadecanol, 1.5% of isopropyl palmitate, 5% of vaseline, 4% of white oil, 5% of silicone oil, 1% of vitamin E, 2% of apocynum venetum essential oil, 2% of emulsifier, 3% of propylene glycol, 5% of apocynum venetum polysaccharide, 3% of nano titanium dioxide, 1% of triethanolamine, 0.5% of phenoxyethanol and the balance of deionized water.

3. The use according to claim 2, wherein the preparation method of the cosmetic with the effects of resisting ultraviolet radiation, resisting oxidation and whitening simultaneously comprises the following steps:

and mixing the oil phase system, the water phase system, the apocynum venetum essential oil and the phenoxyethanol to obtain the cosmetic with the ultraviolet radiation resisting, oxidation resisting and whitening effects.