阅读说明：本技术 一种决明子复方胶原脂质体及其应用 (Cassia seed compound collagen liposome and application thereof ) 是由 李明媛 李萌 李世勤 贾琳 李媛 郁彭 于 2021-06-10 设计创作，主要内容包括：本发明公开一种以决明子复方胶原脂质体为主的制备方法及其应用。一种决明子复方胶原脂质体,由以下原料按质量百分比制成：复方卵磷脂2.00-5.00％、决明子提取液12.00-25.00％、薄荷提取液12.00-25.00％、石斛提取液12.00-25.00％、荸荠提取液12.00-25.00％、红枸杞提取液12.00-25.00％、甘草提取液12.00-25.00％、胶原蛋白溶液8.00-16.00％。本发明以决明子复方胶原脂质体为主的应用兼具明目、保湿、美白、抗皱和高渗透的作用,且质地温和清爽,能够有效缓解眼疲劳,对眼睛的明目作用最为明显。(The invention discloses a preparation method and application of a cassia seed compound collagen liposome. The compound semen cassiae collagen liposome is prepared from the following raw materials in percentage by mass: 2.00-5.00% of compound lecithin, 12.00-25.00% of cassia seed extract, 12.00-25.00% of mint extract, 12.00-25.00% of dendrobe extract, 12.00-25.00% of water chestnut extract, 12.00-25.00% of lycium ruthenicum extract, 12.00-25.00% of liquorice extract and 8.00-16.00% of collagen solution. The application of the cassia seed compound collagen liposome as the main material has the effects of improving eyesight, preserving moisture, whitening, resisting wrinkles and high permeability, is mild and fresh in texture, can effectively relieve eye fatigue, and has the most obvious effect of improving eyesight.)

1. The compound cassia seed collagen liposome is characterized in that: the composite material is prepared from the following raw materials in percentage by mass: 2.00-5.00% of compound lecithin, 12.00-25.00% of cassia seed extract, 12.00-25.00% of mint extract, 12.00-25.00% of dendrobe extract, 12.00-25.00% of water chestnut extract, 12.00-25.00% of lycium ruthenicum extract, 12.00-25.00% of liquorice extract and 8.00-16.00% of collagen solution.

2. The method for preparing the cassia seed compound collagen liposome according to claim 1, which is characterized by comprising the following steps:

(1) dissolving compound lecithin in a small amount of absolute ethyl alcohol, and performing rotary evaporation under reduced pressure to form a film;

(2) preparing hydration solution containing semen Cassiae extract, herba Menthae extract, herba Dendrobii extract, corm Eleocharitis extract, fructus Lycii extract, Glycyrrhrizae radix extract and collagen solution at certain concentration, filtering, and preheating in water at certain temperature;

(3) under the same conditions, adding a hydration solution for hydration, carrying out ultrasonic treatment after complete dissolution until the solution is uniform and clear, and carrying out filtration sterilization to obtain the cassia seed compound collagen liposome.

3. The method for preparing the cassia seed compound collagen liposome according to claim 2, which is characterized in that: the compound lecithin in the step (1) comprises egg yolk lecithin, anhydrous cream and cholesterol, and the mass ratio of the egg yolk lecithin to the anhydrous cream is (3-6): 3: 1; the mass ratio of the compound lecithin to the cassia seed extracting solution is (3-5): 100.

4. the method for preparing the cassia seed compound collagen liposome according to claim 2, which is characterized in that: the hydration liquid in the step (2) comprises 5-10 mg/mL of cassia seed extract, mint extract, dendrobium extract, water chestnut extract, red wolfberry extract and liquorice extract, and 1-5 mg/mL of collagen solution.

5. The method for preparing the cassia seed compound collagen liposome according to claim 2, which is characterized in that: the preheating temperature in the step (2) is 45-50 ℃.

6. The method for preparing the cassia seed compound collagen liposome according to claim 2, which is characterized in that: the filtration in the steps (2) and (3) is filtration by using a 0.22 μm filtration membrane.

7. The application of the cassia seed compound collagen liposome of claim 1, which is embodied as eye cream, eye patch, eye essence and other eye special skin care products, and is characterized in that: 40.00-55.00% of cassia seed compound collagen liposome, 1.00-3.00% of cassia seed extract, 0.20-0.50% of sodium benzoate, 0.50-1.00% of lycium ruthenicum extract, 0.50-1.00% of selfheal extract, 0.50-1.00% of butterflybush flower extract, 0.50-1.00% of glossy privet fruit extract, 0.50-1.00% of water chestnut extract, 0.50-1.00% of ash bark extract, 0.50-1.00% of pipewort extract, 0.50-1.00% of equisetum extract, 0.40-0.80% of arbutin, 0.10-0.30% of tea polyphenol, 0.05-0.10% of hyaluronic acid, 0.05-0.08% of hesperetin, 0.80-2.00% of nicotinamide, 3.00-5.00% of trehalose, 3.00-7.00% of glycerol, 5.00-10.00% of xanthan gum and 30.00-50.00% of sterile distilled water.

8. The preparation method of the application of the cassia seed compound collagen liposome according to claim 7, which is characterized in that: the method comprises the following steps:

(1) weighing raw materials of xanthan gum, hyaluronic acid, trehalose, glycerol and sodium benzoate according to the mass percentage, mixing and placing on a magnetic stirrer, setting a proper rotating speed and temperature, dissolving and stirring uniformly to obtain a basic matrix;

(2) weighing raw materials of cassia seed extract, selfheal extract, buddleja officinalis extract, glossy privet fruit extract, water chestnut extract, ash bark extract, pipewort extract, horsetail extract, sterile distilled water, tea polyphenol, hesperetin and nicotinamide according to mass percentage under the same condition, adding the raw materials in the step (1), dissolving and stirring uniformly;

(3) and (3) reducing the temperature to a certain temperature, weighing the compound collagen liposome and arbutin of the raw materials of the cassia seed according to the mass percentage, adding the compound collagen liposome and the arbutin into the raw materials in the step (2), and stirring the mixture uniformly to obtain a jelly-shaped solid, namely the special eye skin care product.

9. The preparation method of the application of the cassia seed compound collagen liposome according to claim 8, which is characterized in that: the rotating speed in the step (1) is set to be 800-1800 rpm.

10. The preparation method of the application of the cassia seed compound collagen liposome according to claim 8, which is characterized by comprising the following steps: the temperature in the step (1) is set to be 70-80 ℃, and the temperature in the step (3) is set to be 40-45 ℃.

Technical Field

The invention relates to the technical field of liposome and application thereof, in particular to a preparation method mainly based on cassia seed compound collagen liposome and application thereof.

Background

The eye skin of a human is weak, is more easily damaged by the external environment, and is one of the most aging parts. The aging is embodied in the eye skin as the decrease of skin thickness, the decrease of water content, the loose skin, the increase of fine wrinkles, dark circles, the pigmentation and darkness of the skin around the eyes, and the essence is the decrease of the oxidation resistance of the skin, the accumulation of lipofuscin, the decrease of the synthesis capacity of collagen, the decrease of hyaluronic acid content, the decrease of the water content of the skin and the like.

Meanwhile, more and more people always feel dry eyes, sour and swollen eyes, pain and blurred vision even cause visual deterioration because of visual fatigue caused by long-time use of computers, television, book reading, office work and study under the air-conditioning environment and the like.

In order to solve the two problems, most people apply eye cream or other skin care products with corresponding functions when the eye skin is not aged or the aging symptoms appear, and drop eye drops to relieve the eye discomfort. If problems occur at the same time, the processing is troublesome. Therefore, if a special eye skin care product with the functions of moisturizing, whitening and resisting wrinkles can be developed, the eye fatigue can be effectively relieved when the eyes are not comfortable, the eye skin care product has the effects of improving eyesight and protecting the eyes, and the special eye skin care product has a huge market prospect.

Lycium ruthenicum Murr, a dry mature fruit of Lycium barbarum L belonging to Solanaceae, is a traditional and famous Chinese medicinal material in China, has a long medicinal history, is listed as the top grade in Shennong's herbal Jing, and is recorded in Shen nong herbal compendium, which is characterized in that: medlar, tonifying kidney and producing essence, nourishing liver, improving eyesight, strengthening essence and bone, relieving fatigue, facilitating color, whitening, improving eyesight and calming nerves, and prolonging life, has great medicinal and health-care value in the promotion of past doctors. The medlar is mainly distributed in Qinghai, Ningxia, Gansu, Xinjiang and other places in northwest of China. The medlar is used as the traditional Chinese medicinal material and has the functions of nourishing liver and kidney, replenishing vital essence and improving eyesight. Modern pharmacological research proves that the traditional Chinese medicine composition has the effects of regulating immunity, protecting liver, resisting oxidative damage, protecting vision and the like.

Prunellae Spica, a plant of Prunella L of Labiatae, distributed in Henan, Zhejiang and Anhui of China, is recorded in Shen nong Ben Cao Jing, listed below and recorded with "main cold and heat, scrofula, fistula, head sore, broken symptom, goiter and stagnation of qi, and foot swelling and damp arthralgia"; the Danxi Heart method is called nourishing blood vessel; yunnan materia Medica (Yunnan materia Medica) says "dispelling liver wind and moving meridians and collaterals. For facial paralysis, it can relieve arthralgia and myalgia, dredge liver-qi and relieve liver stagnation. For swelling and pain of eyes, it can resolve scrofula. "the ancient people have studied the efficacy and indications of Prunella vulgaris deeply. Spica Prunellae is pungent, bitter and cold in flavor, enters liver and gallbladder channels, has effects of clearing pathogenic fire, improving eyesight, resolving hard mass and detumescence, and can be used for treating eye bulbar nociception, headache giddiness, scrofula, goiter, etc. Modern pharmacological research shows that the selfheal has good curative effects of resisting tumors, resisting inflammation, immunizing, resisting oxidation, reducing blood sugar, reducing blood fat, reducing blood pressure, improving eyesight and the like.

Flos Buddlejae, also known as flos Buddlejae, Oryza Glutinosa, flos Buddlejae, is dried bud and inflorescence of flos Buddleja officinalis (Buddleja officinalis Maxim.) belonging to Buddleja of Loganiaceae, and is collected in spring when the flower is not opened. It is sweet in nature and slightly cold in nature, enters kidney meridian, has effects of clearing heat-fire, nourishing liver to improve eyesight, and eliminating nebula, and can be used for improving eyesight, eliminating nebula, conjunctival congestion, swelling and pain, lacrimation, photophobia, etc. It is well documented by many ancient Chinese medical writings. Beginning with Kai Bao Ben Cao (materia Medica of Kai Bao): mainly has the effects of eliminating skin nebula, astringing, discharging more eye secretion, eliminating red pulse in eyes, eliminating infantile bran beans and infantile malnutrition, and is defined as a special ophthalmic medicine; the records in Ben Cao Jing Shu (herbal Jing Shu): the medicine is sweet and can enrich the blood, remove heat due to cold, and ensure that liver blood is sufficient and the symptoms are not cured ; record of materia medica asking for truth: pale butterflybush flower, which is thin in flavor and thin in flavor, is used with the blood-nourishing herbs to make it stronger.

Fructus Ligustri Lucidi is dry mature fruit of Ligustrum lucidum Ait (Ligustrum lucidum Ait) belonging to Oleaceae. It is sweet, bitter and cool in nature and flavor, and enters liver and kidney meridians. Fructus Ligustri Lucidi has effects of strengthening body resistance, consolidating constitution, nourishing liver and kidney, improving eyesight, and blackening hair. The book listed in Shen nong Ben Cao Jing is the top grade, which is called 'bitter and neutral taste, tonifying middle-jiao, calming five internal organs, nourishing spirit and removing all diseases'. Li Shizhen is said to "fruit is capable of strengthening yin, strengthening waist and knee, whitening hair and improving eyesight".

Water chestnuts are commonly called as water chestnuts, also called as Chinese chestnuts and black tea, are water chestnuts (Eleocharis dulcis (Burm.f.) Trin.) of the family Cyperaceae) and belong to perennial herbaceous plants, and underground stems of the water chestnuts are widely planted in southern provinces of Yangtze river in China. The water chestnut takes underground corms as edible organs, is a food with homology of medicine and food, has tender meat quality, multiple tastes and sweet, is crisp and tasty, has the reputation of underground snow pears, and is regarded as the ginseng in south of the river in the north. According to the record of Chinese medicine dictionary: the water chestnut is sweet in nature and taste, slightly cold and non-toxic, and has the effects of warming the middle-jiao and replenishing qi, clearing heat and stimulating appetite, and helping digestion and reducing phlegm. According to the traditional Chinese medicine, the water chestnuts are sweet and cold in nature and have the health-care effects of clearing heat, reducing phlegm, promoting the production of body fluid, stimulating appetite, improving eyesight, clearing sound, promoting digestion, sobering up and the like.

The cortex fraxini is dry branch bark or dry bark of Fraxinus rhynchophylla Hance (Fraxinus rhynchophylla Hance), Fraxinus chinensis Roxb (Fraxinus chinensis Roxb.), Fraxinus acuminata Lingelsh (Fraxinus szaboana Lingelsh.), Fraxinus chinensis Lingelsh (Fraxinus styrylosa Lingelsh.), and Fraxinus chinensis Lingelsh (Fraxinus styrylosa Lingelsh.), which are commonly used in clinic and are listed as middle-quality products from Shennong Ben Cao Jing. The cortex Fraxini contains coumarin compounds as main chemical components, phenols, saponins, tannin, alkaloids, etc., has effects of clearing heat, eliminating dampness, relieving asthma, relieving cough, and improving eyesight, and can be used for treating enteritis, leukorrhagia, chronic bronchitis, bacillary dysentery, conjunctival congestion, swelling and pain, epiphora induced by wind, psoriasis, etc. The ash bark is widely distributed in China and rich in resources, and is a medicinal plant with high development and utilization values.

Pipewort, which is a head-like inflorescence of a dry flower stem of a pipewort (Eriocaulon buergerianum korn.) plant, widely distributed in sandy ponds or swamps in tropical and subtropical regions of the world, and is abundant in tropical zones in south america; in China, the oryziaceae grows mostly in the south and southwest areas, and the growth environment is mostly damp such as swamps and rice fields. Gujing Cao is recorded in the Song Dynasty 'Kaibao Ben Cao' recorded in Kaibao Biao for treating sore and scabies. The compendium of materia medica records that the medicine is pungent, sweet and neutral in flavor, and mainly treats headache, nebula membrane of blindness and nebula after acne and stops bleeding. The modern Chinese medicine holds that the pipewort has the effects of dispelling wind, dissipating heat, improving eyesight and removing nebula.

Common scouring rush herb, also called as Equisetum hyemale L, is the whole herb of Equisetaceae plants, and is mainly produced in northeast, Shaanxi, Hubei, etc. Scouring rush is first recorded in Jiayou materia Medica, has sweet and slightly bitter taste and mild nature, and has the effects of dispelling wind and dissipating heat, improving eyesight and removing nebula and stopping bleeding. Equisetum hybridum from compendium of materia medica has the functions of dispelling wind, dissipating heat, relieving muscles and removing nebula; equisetum arvense can treat "pain in the main node due to qi tumor and" urgent ascending qi ". Modern medical research proves that the common scouring rush herb has the functions of dispelling wind heat, removing nebula, eliminating masses, benefiting liver and gallbladder, treating intestinal wind, stopping bleeding, removing rheumatism, calming and resisting convulsion and the like.

Cassia seed, which is dry mature seed of Cassia obtusifolia L or Cassia tora L, is recorded in Shennong's herbal Benjing and listed as the top grade. The compendium of materia medica indicates that cassia seed is recorded for treating liver and gallbladder wind heat, skin obstruction, white membrane and cyanosis, and the justice of materia medica also records that cassia seed is used for improving eyesight, nourishing liver and kidney, and relieving diving and nourishing yin, which is the justice of banking up root. Modern researches show that the cassia seed has the effects of reducing blood pressure, reducing blood fat, protecting liver, improving eyesight, resisting oxidation, inhibiting bacteria and the like. As a medicine and food dual-purpose product, the cassia seed has obvious effects on clinical disease prevention and treatment, health preservation and the like, and meanwhile, the cassia seed is widely distributed in China and has low price, so the cassia seed has better development and utilization prospects.

Dendrobium nobile is a plant belonging to the genus Dendrobium nobile (Dendrobium nobile Lindl) of the family Orchidaceae, is one of the traditional famous traditional Chinese medicines in China, also called Dendrobium nobile, Panicum nobile, Panaxanthus diffusa and the like, and the stem thereof is one of the sources of the traditional Chinese medicine, namely Dendrobium nobile, and is collected in the Chinese pharmacopoeia of the calendar edition. Dendrobe is recorded as early as Shen nong Ben Cao Jing, and Ben Cao gang mu says that it has the effects of nourishing yin, benefiting essence, thickening intestines and stomach, tonifying internal spore deficiency, reducing weight and prolonging life. All the herbs in all the past generations such as Ben Cao Yan Yi (the Yan Yi of materia Medica), Ben Cao gang mu Shi Yi (the supplement of compendium of materia Medica) and Ben Cao Shi Xin (the supplement of materia Medica) are called as Qian jin Cao. The dendrobium is slightly cold in nature and sweet in taste, is used for treating body fluid impairment due to heat diseases, dry mouth, polydipsia, asthenic fever after diseases, dim and unclear eyes and the like, and is an important compatibility of preparations such as dendrobium noctilucent pills, dendrobium eyesight improving pills, dendrobium extract solution, dendrobium stomach-clearing powder and the like. In recent years, researches show that dendrobium nobile lindl has remarkable effects on resisting tumors, reducing blood sugar and the like.

Herba Menthae is dry aerial part of Mentha haplocalyx Briq (Mentha haplocalyx Briq) belonging to Labiatae, mainly produced in Jiangsu, Zhejiang and Hunan provinces, and Taicang of Jiangsu is a region of origin, and has effects of dispelling wind and heat, clearing heat, relieving exterior syndrome, relieving sore throat, promoting eruption, dispelling pathogenic wind, relieving swelling, relieving sore throat, and relieving pain, and can be used for treating wind-heat type common cold, headache, conjunctival congestion, pharyngitis, aphtha, rubella, measles, chest and hypochondrium swelling and distress. Mint was originally recorded in Tang Ben Cao (herbal Tang materia Medica) to treat wind, sweating, nausea, abdominal distention, cholera, indigestion and qi descending. Mention is made in Bencao gang mu: to relieve sore throat and relieve sore throat. For scrofula, sores, scabies, wind-induced pruritus and urticaria. The efficacy of mint carried in pharmacopoeia of the people's republic of China is to disperse wind and heat, clear head and eyes, relieve sore throat and promote eruption, and sooth liver and destroy qi. The herba Menthae is rich in chemical components, and mainly contains volatile oil, flavonoids, anthraquinones, organic acids, amino acids, microelements, etc. Modern pharmacological research shows that the mint has the effects of protecting liver, benefiting gallbladder, exciting human central nerve, resisting inflammation, easing pain, resisting virus, promoting transdermal absorption and the like.

The Glycyrrhrizae radix is dried root and rhizome of Glycyrrhiza uralensis Fisch, Glycyrrhiza glabra L or Glycyrrhiza inflata Bat of Leguminosae. Mainly comes from inner Mongolia, Gansu, Xinjiang and other places, and has the effects of tonifying qi, detoxifying, eliminating phlegm, relieving cough, clearing heat, relieving pain and the like. Licorice root, radix Glycyrrhizae has the action of "relieving Baicao-du", and usually has the actions of removing toxicity and harmonizing the effects of other drugs in the formula.

Collagen, one of the most important structural proteins in the organism, accounts for 25 to 30 percent of the total protein content, is mainly present in skin, bones and ligaments, and plays a role in supporting and protecting the organism. The collagen in the skin of animals is mainly white transparent type I fibrous collagen with quaternary structure. Collagen is closely related to tissue formation, maturation, intercellular information transmission, joint lubrication, wound healing, calcification, blood coagulation, aging, etc. Collagen is one of the most critical raw materials in the biotechnology industry, and is widely applied to medical materials, cosmetics and food industries. Collagen has remarkable advantages in cosmetics, and is widely used in cosmetics due to its characteristics of good biocompatibility, biodegradability and bioactivity, low antigenicity, easy absorption by the human body, promotion of cell survival and growth, promotion of platelet aggregation, and the like.

Liposomes, bilayer vesicles composed of phospholipids as the major material, were first discovered by british scientists to be structurally similar to biological membranes. In the last 70 s, liposomes began to become drug carriers. Due to the advantages of biocompatibility, biodegradability, non-toxicity and non-immunogenicity, the liposome is rapidly developed as a new medicament form, and 11 liposome preparations are on the market at present. In the research of transdermal effect, research shows that the liposome with the particle size of 20-200 nm can be used as an active carrier for local administration and can permeate into the epidermis of a living body by means of intercellular space. The liposome in cosmetics has the advantages of high permeability, light, heat and oxidation instability protection of effective components, activity maintenance and stabilization, and can enter into skin deep cells through stratum corneum gap.

The cosmetics containing the cassia seeds are fresh at home and abroad, and the cassia seeds are prepared into the form of the compound collagen liposome of the cassia seeds due to low skin absorption efficiency of the cassia seeds, so that the medicine can be continuously released at the skin to increase the medicine concentration, thereby obviously increasing the medicine penetration amount, being beneficial to increasing the medicine concentration of local skin, reducing adverse reactions such as medicine stimulation and the like, and being beneficial to local application of the skin. Therefore, the inventor develops the special eye skin care product containing the cassia seed compound collagen liposome.

The invention relates to a Chinese invention patent (patent application No. 201711253559.5, publication No. CN107929198A, application date: 2017-12-02), the invention name of which is 'eye cream formula', discloses an eye cream, which is composed of glycyrrhizic acid concentrated powder, rose essential oil, cassia seed powder, arnica montana extract and the like, and is obviously different from the formula of the invention.

The invention relates to Chinese patent (patent application No. 201610462469.6, publication No. CN 107519338A, application date: 2016-06-22), the name of the invention is 'a Chinese medicinal composition for external use for protecting eyesight', and discloses an eye cream. Clinical tests show that it has obvious therapeutic effect for visual fatigue of visual disturbance, eye sleepiness, aching pain, lacrimation and foreign body sensation, etc.. But the required raw materials have more components, the preparation and the production are complicated, the medicine needs to be used together with instruments, and the treatment is mainly performed, so the protection and the prevention are not clear.

The invention relates to an anti-aging eye cream and a preparation method thereof, and discloses an eye cream, wherein the eye cream is added with a plurality of plant active ingredients, and the main plant active ingredients are Cannabidiol (CBD) and astaxanthin, which are obviously different from the main ingredients of the invention (patent application No. 201911322832.4, publication No. CN 110787113A, application date: 2019-12-20).

The invention adopts the form of cassia seed compound collagen liposome, is compatible with other medicinal and edible traditional Chinese medicine plant effective active ingredients, has the synergistic effect among the ingredients, has the effects of promoting qi and activating blood, clearing liver and improving eyesight, can promote the blood circulation of the skin around eyes, enhance the activity of cells, repair damaged cells and provide rich nutrient components for the damaged cells, deeply moisten the skin around eyes, delay the aging of the skin around eyes, improve the existing fine lines, keep moisture and whiten the skin well, has obvious eyesight improving effect and safe and comfortable use, is safe and comfortable to use, is a natural skin supporting agent added in the formula, has high safety, is mild and non-irritating, can directly reach the skin around eyes by a liposome targeting technology, has higher oral administration utilization rate and more concentrated distribution, has strong affinity to the skin around eyes and good biocompatibility, and enables the skin around eyes to achieve a good oil-water balance effect, better absorption of nutrients.

Disclosure of Invention

Based on the above mentioned problems, the invention provides a special eye skin care product mainly comprising cassia seed compound collagen liposome, which can simultaneously prevent and solve the eye skin aging and discomfort caused by long-term eye use, and has the function of improving eyesight. The invention also takes liposome as a carrier, which not only increases the solubility of the medicine components, but also improves the stability of the medicine and the bioavailability of the medicine.

The invention aims to provide a preparation method of the cassia seed compound collagen liposome.

The technical scheme adopted by the invention is that the formula of the cassia seed compound collagen liposome is characterized in that: the composite material is prepared from the following raw materials in percentage by mass: 2.00-5.00% of compound lecithin, 12.00-25.00% of cassia seed extract, 12.00-25.00% of mint extract, 12.00-25.00% of dendrobe extract, 12.00-25.00% of water chestnut extract, 12.00-25.00% of lycium ruthenicum extract, 12.00-25.00% of liquorice extract and 8.00-16.00% of collagen solution.

The invention also aims to provide the application of the cassia seed compound collagen liposome as the main material, in particular to a preparation method of the special eye skin care product.

The second technical scheme adopted by the invention is an application of cassia seed compound collagen liposome as a main material, which is specifically expressed as a skin care product special for eyes, and is characterized in that: 40.00-55.00% of cassia seed compound collagen liposome, 1.00-3.00% of cassia seed extract, 0.20-0.50% of sodium benzoate, 0.50-1.00% of lycium ruthenicum extract, 0.50-1.00% of selfheal extract, 0.50-1.00% of butterflybush flower extract, 0.50-1.00% of glossy privet fruit extract, 0.50-1.00% of water chestnut extract, 0.50-1.00% of ash bark extract, 0.50-1.00% of pipewort extract, 0.50-1.00% of equisetum extract, 0.40-0.80% of arbutin, 0.10-0.30% of tea polyphenol, 0.05-0.10% of hyaluronic acid, 0.05-0.08% of hesperetin, 0.80-2.00% of nicotinamide, 3.00-5.00% of trehalose, 3.00-7.00% of glycerol, 5.00-10.00% of xanthan gum and 30.00-50.00% of sterile distilled water.

The third technical scheme adopted by the invention is that the preparation method of the cassia seed compound collagen liposome comprises the following steps:

dissolving compound lecithin in a small amount of absolute ethyl alcohol, and performing rotary evaporation under reduced pressure to form a film;

step two, preparing hydration liquid with certain concentration and containing cassia seed extract, mint extract, dendrobium extract, water chestnut extract, red medlar extract, liquorice extract and collagen solution, filtering, and putting into water with certain temperature for preheating;

and step three, adding the hydration liquid into the mixture under the same conditions, carrying out ultrasonic treatment after complete dissolution until the solution is uniform and clear, and carrying out filtration sterilization to obtain the cassia seed compound collagen liposome.

The present invention is also characterized in that,

the compound lecithin in the first step comprises yolk lecithin, anhydrous cream and cholesterol, and the mass ratio of the yolk lecithin to the anhydrous cream to the cholesterol is (3-6): 3: 1; the mass ratio of the compound lecithin to the cassia seed extracting solution is (3-5): 100.

the hydration liquid in the step one comprises 5-10 mg/mL of cassia seed extract, mint extract, dendrobium extract, water chestnut extract, red wolfberry extract and liquorice extract, and 1-5 mg/mL of collagen solution.

And the preheating temperature in the second step is 45-50 ℃.

And the filtration in the second and third steps is filtration by using a 0.22 mu m filter membrane.

The invention adopts the fourth technical scheme that a preparation method mainly using cassia seed compound collagen liposome as a main material, in particular to a preparation method of a special eye skin care product, which comprises the following steps:

weighing raw materials of xanthan gum, hyaluronic acid, trehalose, glycerol and sodium benzoate according to the mass percentage, mixing and placing the raw materials on a magnetic stirrer, setting the proper rotating speed and temperature, and dissolving and stirring the raw materials uniformly to obtain a basic matrix;

step two, weighing raw materials of cassia seed extract, selfheal extract, buddleja officinalis extract, glossy privet fruit extract, water chestnut extract, ash bark extract, pipewort extract, horsetail extract, sterile distilled water, tea polyphenol, hesperetin and nicotinamide according to mass percentage under the same condition, adding the raw materials in the step one, dissolving and stirring uniformly;

and step three, reducing the temperature to a certain temperature, weighing the compound collagen liposome and arbutin of the raw materials of the cassia seed according to the mass percentage, adding the compound collagen liposome and the arbutin into the raw materials of the step two, and stirring the mixture uniformly to obtain a jelly-shaped solid, namely the special eye skin care product.

The present invention is also characterized in that,

the rotating speed in the first step is set to be 800-1800 rpm, and the temperature is set to be 70-80 ℃.

The temperature in the third step is set to be 40-45 ℃.

Compared with the prior art, the invention has the advantages that,

according to the invention, the cassia seed compound collagen liposome has a certain antioxidation effect on eye skin, has a certain eyesight improving effect on eye cells, has a moisturizing effect on hyaluronic acid and trehalose, the wolfberry extract has the effects of nourishing yin, enriching blood, improving eyesight and nourishing liver, the hesperetin has the effects of activating blood and resisting oxidation, the arbutin and the nicotinamide have the effects of whitening and the like, a unique formula is innovatively formed, multiple functions of improving eyesight, whitening, moisturizing, resisting wrinkles, removing edema and the like are considered in the formula, and discomfort caused by long-time eye use can be relieved, so that the comprehensive nursing effect is achieved, and convenience is brought to life of people.

The special eye skin care product using the cassia seed compound collagen liposome as the carrier innovatively adopts the cassia seed compound collagen liposome as the carrier of the effective component, can realize transdermal delivery of the effective component, and improves the stability of the effective component. The invention utilizes liposome technology to increase the solubility of the effective components and improve the bioavailability of the effective components.

Thirdly, the formula of the cassia seed compound collagen liposome is added with a natural skin propping agent, namely collagen, which is mainly present in skin, bones and ligaments and plays a role in supporting and protecting organisms. It has biodegradability and bioactivity, low antigenicity, easy absorption by human body, and is widely used in cosmetics to promote cell survival and growth and platelet coagulation. The invention can directly reach eye skin by a liposome targeting technology, has higher utilization rate and more concentrated distribution compared with oral administration, has strong affinity of collagen to the eye skin, has good biocompatibility, can effectively reduce stimulation, reduces potential stimulation of other surfactants, and can better care delicate skin around eyes.

The traditional Chinese medicine raw materials selected by the invention are mostly medicinal and edible medicinal materials, so that the traditional Chinese medicine composition is high in safety and small in toxic and side effects, and on the basis of the formula theory of the traditional Chinese medicine, the medicines have the effects of protecting liver and improving eyesight. In addition, the traditional Chinese medicines selected by the invention have wide sources, low price, clear and various effects and large available space, so that the preparation cost is low, the preparation process is easy for industrial production, the application range is wide, and the traditional Chinese medicine can be accepted by more people.

Drawings

FIG. 1 is a photograph of a three-cassia seed compound collagen liposome of the example;

the cassia seed compound collagen liposome solution is yellow brown clear translucent uniform liquid without layering and precipitation.

FIG. 2 is a diagram showing the particle size distribution of the cassia seed compound collagen liposome of the third embodiment;

the average particle diameter of the cassia seed compound collagen liposome is 95nm, the average particle diameter is 70-100 nm, the distribution is concentrated and uniform, and the cassia seed compound collagen liposome is a good nano dispersion system.

FIG. 3 is a bovine serum albumin standard curve.

FIG. 4 shows the results of tyrosinase inhibition experiments at different concentrations in example VII, which shows that example VII has good antioxidant effect.

FIG. 5 shows the result of DPPH radical scavenging experiment of example eight with different concentrations, and the result shows that example eight has good antioxidant effect.

FIG. 6 is a photograph of the state of BV2 cells stimulated with lipopolysaccharide (1. mu.g/mL) at the time of addition of 0.8mg/mL of the example.

FIG. 7 is a photograph of the state of BV2 cells stimulated by lipopolysaccharide (1. mu.g/mL) when 0.6mg/mL of example two was added.

FIG. 8 is a photograph of the state of BV2 cells stimulated with the addition of lipopolysaccharide alone (1. mu.g/mL).

FIG. 9 shows the results of experiments on the survival rate of BV2 stimulated by lipopolysaccharide (1. mu.g/mL) of the first and second examples at different concentrations, which shows that the second example has better protection effect than the first example.

FIG. 10 is a photograph showing the state of BV2 cells stimulated with hydrogen peroxide alone (20. mu. mol/mL).

FIG. 11 shows the results of experiments on the survival rate of BV2 stimulated by hydrogen peroxide solution (1mmol/mL) in examples one and two at different concentrations, which shows that the protection effect of example two is better than that of example one.

FIG. 12 shows the results of experiments on how much different concentrations of the first and second protective hydrogen peroxide solutions (20. mu. mol/mL) stimulate BV2 cell viability, and shows that the second protective solution has better protective effect than the first protective solution.

Fig. 13 is a photograph showing the state of BV stimulating 2 cells when only blue light is irradiated (irradiation time 35 min).

FIG. 14 shows the results of experiments on the survival rate of BV2 stimulated by blue light (irradiation time 35min) in different concentrations for example one and example two, which shows that example two has better protection effect than example one.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following examples, which are illustrative of the present invention and the description thereof, and are not to be construed as limiting the present invention.

Comparative example 1

A vitamin C solution having a concentration of 1mg/mL was prepared using ultrapure water, and the vitamin C solution was diluted with ultrapure water to 0.8mg/mL, 0.6mg/mL, 0.4mg/mL, and 0.2 mg/mL.

Comparative example No. two

Preparing a vitamin C solution with the concentration of 1mg/mL by using absolute ethyl alcohol, and diluting the vitamin C solution into 0.8mg/mL, 0.6mg/mL, 0.4mg/mL and 0.2mg/mL by using the absolute ethyl alcohol.

Example one

Preparing semen Cassiae extract with 10mg/mL MEM medium containing diabody and serum, filtering, sterilizing, and diluting with 8mg/mL, 6mg/mL, 4mg/mL, 2mg/mL MEM medium containing diabody and serum.

Example two

Dissolving compound lecithin in a small amount of absolute ethyl alcohol, and performing rotary evaporation under reduced pressure to form a film; preparing 10mg/mL hydration solution containing semen Cassiae extract, filtering, and preheating in 50 deg.C water; adding water into the liquefaction solution for hydration under the same conditions, carrying out ultrasonic treatment after complete dissolution until the solution is uniform and clear, and carrying out filtration sterilization to obtain the cassia seed liposome.

The compound lecithin described in the second embodiment comprises egg yolk lecithin, anhydrous cream and cholesterol, and the mass ratio of the egg yolk lecithin to the anhydrous cream is 3: 3: 1.

EXAMPLE III

Dissolving compound lecithin in a small amount of absolute ethyl alcohol, and performing rotary evaporation under reduced pressure to form a film; preparing 10mg/mL hydration solution containing semen Cassiae extract, herba Menthae extract, herba Dendrobii extract, corm Eleocharitis extract, fructus Lycii extract, Glycyrrhrizae radix extract and 5mg/mL collagen solution, filtering, and preheating in 50 deg.C water; under the same conditions, adding a hydration solution for hydration, carrying out ultrasonic treatment after complete dissolution until the solution is uniform and clear, and carrying out filtration sterilization to obtain the cassia seed compound collagen liposome.

The compound lecithin described in the third embodiment comprises egg yolk lecithin, anhydrous cream and cholesterol, and the mass ratio of the egg yolk lecithin to the anhydrous cream is 3: 3: 1.

example four

Using MEM culture medium containing double antibiotics and serum to prepare 10mg/mL cassia seed extract, honeysuckle extract, water chestnut extract, equisetum hiemale extract, ash bark extract, buddleja officinalis extract, selfheal extract, collagen liposome (the preparation method is the same as that in the first embodiment), vitamin C, cassia seed liposome, licorice extract, pipewort extract, red wolfberry extract, glossy privet fruit extract, mint extract, dendrobium extract, hyaluronic acid extract and gallic acid extract, and filtering and sterilizing.

EXAMPLE five

An aqueous collagen solution of 5mg/mL was prepared using physiological saline.

EXAMPLE six

Dissolving compound lecithin in a small amount of absolute ethyl alcohol, and performing rotary evaporation under reduced pressure to form a film; preparing 5mg/mL of hydration liquid containing collagen, filtering, and preheating in water at 50 ℃; adding a hydration solution to hydrate under the same condition, carrying out ultrasonic treatment after complete dissolution until the solution is uniform and clear, and carrying out filtration sterilization to obtain the collagen liposome.

The compound lecithin described in the sixth embodiment comprises egg yolk lecithin, anhydrous cream and cholesterol, wherein the mass ratio of the egg yolk lecithin to the anhydrous cream to the cholesterol is 3: 3: 1.

EXAMPLE seven

Preparing semen Cassiae extract with concentration of 1mg/mL with ultrapure water, filtering, sterilizing, and diluting with ultrapure water to 0.8mg/mL, 0.6mg/mL, 0.4mg/mL, 0.2 mg/mL.

Example eight

Preparing semen Cassiae extract with concentration of 1mg/mL with anhydrous ethanol, filtering, sterilizing, and diluting with anhydrous ethanol to obtain semen Cassiae extract with concentration of 0.8mg/mL, 0.6mg/mL, 0.4mg/mL, 0.2 mg/mL.

Example nine

Weighing 7% of xanthan gum, 0.1% of hyaluronic acid, 5% of trehalose, 5% of glycerol and 0.2% of sodium benzoate according to the mass percentage, mixing and placing on a magnetic stirrer, setting the rotating speed of 1500rpm and the temperature of 75 ℃, dissolving and stirring uniformly to prepare a basic matrix; weighing 2% of cassia seed extract, 0.5% of selfheal extract, 0.5% of flos buddlejae extract, 0.5% of glossy privet fruit extract, 0.5% of water chestnut extract, 0.5% of ash bark extract, 0.5% of pipewort extract, 0.5% of equisetum hiemale extract, 24.95% of sterile distilled water, 0.2% of tea polyphenol, 0.05% of hesperetin and 1.5% of nicotinamide according to mass percentage, adding the raw materials into the raw materials, dissolving and stirring the raw materials uniformly; reducing the temperature to 45 ℃, weighing 50% of the compound collagen liposome of the raw materials of the cassia seed and 0.5% of arbutin according to the mass percentage, adding the compound collagen liposome of the raw materials into the raw materials, and stirring the mixture evenly to prepare the eye cream.

Example ten

Weighing 7% of xanthan gum, 0.1% of hyaluronic acid, 5% of trehalose, 5% of glycerol and 0.2% of sodium benzoate according to the mass percentage, mixing and placing on a magnetic stirrer, setting the rotating speed of 1500rpm and the temperature of 75 ℃, dissolving and stirring uniformly to prepare a basic matrix; weighing 2% of cassia seed extract, 0.5% of selfheal extract, 0.5% of flos buddlejae extract, 0.5% of glossy privet fruit extract, 0.5% of water chestnut extract, 0.5% of ash bark extract, 0.5% of pipewort extract, 0.5% of equisetum hiemale extract, 29.95% of sterile distilled water, 0.2% of tea polyphenol, 0.05% of hesperetin and 1.5% of nicotinamide according to mass percentage, adding the raw materials into the raw materials, dissolving and stirring the raw materials uniformly; reducing the temperature to 45 ℃, weighing 45% of the compound collagen liposome of the raw materials of the cassia seed and 0.5% of arbutin according to the mass percentage, adding the mixture into the raw materials, and stirring the mixture evenly to prepare the eye cream.

EXAMPLE eleven

Weighing 7% of xanthan gum, 0.1% of hyaluronic acid, 5% of trehalose, 5% of glycerol and 0.2% of sodium benzoate according to the mass percentage, mixing and placing on a magnetic stirrer, setting the rotating speed of 1500rpm and the temperature of 75 ℃, dissolving and stirring uniformly to prepare a basic matrix; weighing 2% of cassia seed extract, 0.5% of selfheal extract, 0.5% of flos buddlejae extract, 0.5% of glossy privet fruit extract, 0.5% of water chestnut extract, 0.5% of ash bark extract, 0.5% of pipewort extract, 0.5% of equisetum hiemale extract, 34.95% of sterile distilled water, 0.2% of tea polyphenol, 0.05% of hesperetin and 1.5% of nicotinamide according to mass percentage, adding the raw materials into the raw materials under the same condition, dissolving and stirring the raw materials uniformly; reducing the temperature to 45 ℃, weighing 40 mass percent of the cassia seed compound collagen liposome and 0.5 mass percent of arbutin, adding the materials into the raw materials, and stirring the mixture uniformly to prepare the eye cream.

Example twelve

A MEM culture medium containing double antibodies and serum is used for preparing a compound solution with the concentration of 10mg/mL, and the compound solution is specifically divided into the following groups:

(1) herba Dendrobii, HONGGOU, and radix Glycyrrhizae; (2) herba Dendrobii, Glycyrrhrizae radix, and herba Menthae; (3) herba Dendrobii, herba Menthae, and corm Eleocharitis; (4) herba Dendrobii, HONGGOU, and herba Menthae; (5) herba Dendrobii, HONGGOU, and corm Eleocharitis; (6) herba Dendrobii, Glycyrrhrizae radix, and corm Eleocharitis; (7) HONGGOU, Glycyrrhrizae radix, and herba Menthae; (8) HONGGOU, Glycyrrhrizae radix, and corm Eleocharitis; (9) licorice, peppermint, water chestnut; (10) HONGGOU, herba Menthae, and corm Eleocharitis; (11) herba Dendrobii, HONGGOU, Glycyrrhrizae radix, and herba Menthae; (12) herba Dendrobii, Glycyrrhrizae radix, herba Menthae, and corm Eleocharitis; (13) herba Dendrobii, HONGGOU, Glycyrrhrizae radix, and corm Eleocharitis; (14) herba Dendrobii, HONGGOU, herba Menthae, and corm Eleocharitis; (15) HONGGOU, Glycyrrhrizae radix, herba Menthae, and corm Eleocharitis; (16) dendrobe, Chinese wolfberry, liquorice, mint and water chestnut.

Test example-stability test

15g of each of the skin care products for eye exclusive use obtained in examples nine, ten and eleven was sealed and placed in an oven (50. + -.1 ℃ C.), room temperature (25. + -.1 ℃ C.) and a refrigerator (4. + -.1 ℃ C.) for 45 days, and whether the appearance thereof was mildewed, delaminated or precipitated and the absorption state of the skin care product for eye exclusive use on the skin were observed, and the results are shown in Table 1 below.

TABLE 1 stability test results

The results showed that in the high temperature, normal temperature, low temperature environment, the ninth example was significantly delaminated at room temperature, the tenth example was slightly delaminated at high temperature, and the eleventh example was free from delamination, mold, and precipitation, had stable properties, and was well absorbed on the skin and was completely absorbed.

Test example two in vitro transdermal test

Drawing a bovine serum albumin standard curve: completely dissolving 10mg of bovine serum standard substance in 10mL of ultrapure water to prepare 1mg/mL of bovine serum standard substance aqueous solution, and diluting the bovine serum standard substance aqueous solution step by step to prepare 0.5mg/mL, 0.25mg/mL, 0.1mg/mL, 0.01mg/mL, 0.001mg/mL, 0.1 mu g/mL, 0.01 mu g/mL and 0.001 mu g/mL of standard substance solution. And respectively taking 20 mu L of the liquid with each concentration, respectively adding 100 mu L of Coomassie brilliant blue G250 dye solution into a 96-well plate, and measuring the absorbance of the mixture by a 595nm microplate reader. And recording data and drawing a standard curve.

Removing hair from the back skin of C57BL/6 mouse with sodium sulfide and subcutaneous fat, cleaning with warm physiological saline, cutting into appropriate size, cleaning with phosphate buffer (or physiological saline) of pH7.2, soaking for 30min, and blotting surface water with filter paper. The dorsal skin of the treated mice was fixed between a supply chamber and a receiving chamber by using a Valia-Chien diffusion cell, the stratum corneum faced the supply chamber, the supply solutions were example five and example six (3 mL of collagen solution and collagen liposome, respectively, and 2mL of physiological saline), the concentrations of the collagen solution and collagen liposome were 5mg/mL, the receiving solution was physiological saline, the volume was 5mL, and the effective transdermal area was 0.95cm². The device is externally connected with a constant temperature water bath (33 +/-0.1 ℃), a magnetic stirrer is arranged in the device, the rotating speed is 300r/min, and stirring is carried out in the test process. The receiving chamber was withdrawn 200. mu.L of the receiving solution every 1h interval, and immediately supplemented with an equal volume of physiological saline. After the sample is filtered by a 0.45 mu m microporous filter membrane, 20 mu L of the receiving solution is taken and added with 100 mu L of Coomassie brilliant blue G250 dye solution, and the absorbance of the mixture is measured by a 595nm microplate reader.

Calculating the content of collagen: the absorbance value of the sample after measurement is brought into a standard curve linear equation to obtain the concentration of the collagen in the sample, and the concentration of the permeated collagen is detected by a Coomassie brilliant blue method, and the result is shown in the following table 2.

TABLE 2 study of transdermal efficiency of collagen liposomes at different time points

It can be seen from table 2 that the transdermal effect of the collagen liposome is better than that of the collagen solution with the same concentration.

Test example three whitening efficacy evaluation tests (tyrosinase activity inhibition test)

Tyrosinase is the rate-limiting enzyme in the melanin synthesis pathway, and mainly influences the generation of melanin by influencing the conversion of tyrosine into dopa and the oxidation of dopa into dopaquinone, and inhibits the generation of melanin by inhibiting the activity of tyrosinase. Therefore, the efficacy of the inhibition of tyrosinase was evaluated by determining the results of the inhibition of tyrosinase in example seven.

Solution preparation:

preparing 50mL of phosphate buffer solution with the pH value of 6.8 by a conventional preparation method; weighing a proper amount of tyrosine, adding water for dissolving, and preparing 2mmol/L of tyrosine solution and 500U/mL of tyrosinase solution.

A96-well plate was prepared, and 100. mu.L of phosphate buffer solution having pH6.8 and 50. mu.L of the above-mentioned tyrosine solution were added to each well, and 10. mu.L of each of example seven and comparative example 1 was added. After mixing uniformly, 10 μ L of tyrosinase solution was rapidly added in an ice bath, mixed uniformly, incubated at 37 ℃ for 10min, and absorbance was measured at 475 nm.

Tyrosinase inhibition (%) - (A-B) - (C-D) ]/[ A-B ]. times.100%

Wherein, A: absorbance measured without adding the enzyme mixture of example seven; b: absorbance measured in the mixture without the enzyme of example seven; c: adding the absorbance measured by the mixture of example seven and the enzyme; d: the absorbance measured for the mixture containing example seven and no enzyme was measured, and the results are shown in Table 3 below.

Each experimental well has 3 multiple wells, namely n is 3.

TABLE 3 tyrosinase inhibition test results (n ═ 3)

It can be seen from table 3 that the concentration of example seven in the concentration range of 0.2-0.6mg/mL is concentration-dependent on the inhibition of tyrosinase activity, and the whitening ability of example seven is continuously enhanced with the increase of the concentration.

Test example four antioxidant evaluation test (DPPH radical scavenging test)

DPPH radical is a synthetic, one-electron, stable, nitrogen-centered paramagnetic compound. In the presence of the radical scavenger, DPPH radical accepts an electron or hydrogen atom to form a stable DPPH-H compound, which changes its methanol (or ethanol) solution from dark purple to yellow, and the degree of discoloration is quantitative with respect to the number of electrons it accepts (radical scavenging activity), so that rapid quantitative analysis can be performed using a spectrophotometer. Therefore, the present invention evaluates the efficacy of example eight pairs of DPPH radical scavenging results.

Preparing a solution:

weighing 4mg of DPPH free radical powder into a 100mL volumetric flask, dissolving with absolute ethyl alcohol, diluting to a constant volume, and shaking up to obtain 0.04mg/mL DPPH solution.

And (3) taking a 96-well plate, dropwise adding 100 mu L of LDPPH solution, then respectively adding 100 mu L of the eighth example and the second comparative example, uniformly mixing, carrying out a light-shielding reaction for 20min at room temperature, and measuring the ultraviolet absorbance (B) at 515nm, wherein the results are shown in Table 4.

Control group: comparative example No. II, 0.2-1 mg/mL; experimental groups: example eight at 10-50 mg/mL.

DPPH radical scavenging ratio (%) (1-B/A). times.100%

Wherein, A: blank absorbance; b: absorbance of the sample.

The experiment set up 3 parallel test wells, i.e. n-3.

TABLE 4-1 DPPH radical scavenging test results

TABLE 4-2 DPPH radical scavenging test results

As can be seen by comparing tables 4-1 and 4-2, the antioxidant activity of the eighth example is concentration-dependent, and compared with the second comparative example, the eighth example has stable and durable antioxidant capacity and is more suitable for being added into skin care products.

Test example five evaluation tests for efficacy of improving eyesight

The protective effect of example one and example two on BV2 microglia was tested by stimulating BV2 microglia with lipopolysaccharide (1. mu.g/mL), hydrogen peroxide (1mmol/L and 20. mu. mol/L) and blue light (irradiation time 35min) and testing the viability of the cells by MTT method.

MTT method: the detection principle is that succinate dehydrogenase in mitochondria of living cells can enable exogenous MTT to be reduced into water-insoluble blue-violet crystal Formazan (Formazan) and deposited in the cells, and dead cells do not have the function. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and an enzyme linked immunosorbent assay detector is used for measuring the light absorption values at 492nm and 630nm wavelengths, so that the quantity of living cells can be indirectly reflected. Within a certain range of cell number, MTT crystals are formed in an amount proportional to the cell number.

Adding 90 μ L of cell sap into 96-well plate with cell density of 5 × 10⁴Per mL, 37 ℃, 5% CO₂Incubation is carried out for 24 h; adding 10 μ L (1:10) of each of the first and second examples into each well, and stimulating in different ways after 1h (irradiating with lipopolysaccharide, hydrogen peroxide and blue light); adding 20 mu L MTT solution into each hole after 2h, and continuing culturing for 4 h; terminating the culture, adding 100 mu L of isopropanol hydrochloride to dissolve formazan, and incubating for 10 min; measuring the optical density value (OD value) of each hole at the wavelength of 492nm and 630nm on an enzyme-linked immunosorbent detector; the cell viability was calculated and the results are shown in table 5.

Cell viability { (OD value-blank OD value [ mean ])/(blank OD value-blank OD } × 100%

TABLE 5-1a lipopolysaccharide (1. mu.g/mL) stimulation of BV2 cell viability assay results (example one)

Concentration (mg/mL)	0.2	0.4	0.6	0.8	1.0
						Blank control	-0.51	0.03	0.48	1.678	2.14
EXAMPLE I + lipopolysaccharide	82.99	84.73	99.29	106.35	114.50
						EXAMPLE I + lipopolysaccharide	82.08	80.52	107.44	113.95	96.92
EXAMPLE I + lipopolysaccharide	78.32	84.18	95.45	102.59	93.99
						EXAMPLE I + lipopolysaccharide	75.85	88.94	104.05	112.48	100.95
EXAMPLE I + lipopolysaccharide	86.75	90.59	98.56	97.74	108.91
						EXAMPLE I + lipopolysaccharide	90.96	87.66	106.25	120.54	114.77
+ lipopolysaccharide	70.90	86.47	66.69	67.52	62.58
						+ complete culture medium	103.32	94.62	109.64	127.14	96.46

As can be seen from Table 5-1a, the survival rate of cells was the highest at 0.8mg/mL in example one (see FIG. 6), and the cells were also protected and maintained in normal proliferation after the drug addition (the concentration was not toxic to cells, which is the optimum concentration for the cassia seed solution) in comparison with the lipopolysaccharide group (see FIG. 8).

TABLE 5-1b lipopolysaccharide (1. mu.g/mL) stimulation of BV2 cell viability assay results (example two)

Concentration (mg/mL)	1.0	0.8	0.6	0.4	0.2
						Blank control	-1.25	1.59	1.31	-1.01	-0.06
EXAMPLE di + lipopolysaccharide	126.22	130.98	132.26	134.19	122.56
						EXAMPLE di + lipopolysaccharide	126.40	118.99	125.03	128.51	127.69
EXAMPLE di + lipopolysaccharide	118.62	118.62	128.69	120.18	121.28
						EXAMPLE di + lipopolysaccharide	131.99	119.26	127.23	122.47	130.34
EXAMPLE di + lipopolysaccharide	118.25	127.32	142.16	131.17	126.04
						EXAMPLE di + lipopolysaccharide	130.98	132.26	131.44	133.18	130.43
+ lipopolysaccharide	70.91	86.48	66.70	67.52	62.58
						+ complete culture medium	103.33	94.63	109.65	127.14	96.46

As can be seen from tables 5-1b, the cell survival rate was highest at the concentration of 0.6mg/mL in example two (see FIG. 7), and the drug was effective in protecting cells and maintaining cell proliferation (no toxicity to cells at this concentration, optimum for the liposome concentration of Cassia tora), in contrast to the lipopolysaccharide group (see FIG. 8).

TABLE 5-2a Experimental results on the stimulation of BV2 cell viability by hydrogen peroxide (20. mu. mol/L) (example one)

Concentration (mg/mL)	0.2	0.4	0.6	0.8	1.0
						Blank control	1.26	2.16	4.84	-0.53	0.76
Example one + H2O2	26.33	24.14	28.51	33.29	41.45
						Example one + H2O2	20.66	19.86	21.15	19.86	26.82
Example one + H2O2	19.16	21.35	26.33	27.52	31.60
						Example one + H2O2	17.57	22.75	26.53	27.82	33.19
Example one + H2O2	22.05	22.05	27.02	26.82	33.29
						Example one + H2O2	23.74	23.84	27.82	28.51	34.08
+H2O2	18.67	15.68	12.40	13.79	15.38
						+ complete culture medium	93.37	104.41	109.78	102.42	102.12

As can be seen from Table 5-2a, the cell survival rate was highest at 1.0mg/mL in example one, and H₂O₂Compared with the group (as shown in figure 10), the drug has certain protection effect on cells after being added.

TABLE 5-2b Experimental results on the stimulation of BV2 cell viability by hydrogen peroxide (20. mu. mol/L) (example two)

Concentration (mg/mL)	1.0	0.8	0.6	0.4	0.2
						Blank control	1.36	-1.82	-1.13	-2.42	0.96
Example two + H2O2	51.39	39.95	29.31	23.14	9.91
						Example two + H2O2	53.58	37.47	31.20	21.35	13.89
Example two + H2O2	55.87	38.36	25.73	19.86	13.99
						Example two + H2O2	70.49	53.38	35.08	23.24	12.30
Example two + H2O2	63.53	54.38	35.68	23.14	16.98
						Example two + H2O2	65.02	45.42	40.15	32.00	22.25
+H2O2	18.67	15.68	12.40	13.79	15.39
						+ complete culture medium	93.37	104.41	109.78	102.42	102.12

As can be seen from tables 5-2b, the cell viability was highest at 1.0mg/mL for example two, and H₂O₂Compared with the group (as shown in figure 10), the drug has certain protection effect on cells after being added.

TABLE 5-2c experimental results of BV2 cell viability stimulation by hydrogen peroxide (1mmol/L) (example I)

Concentration (mg/mL)	0.2	0.4	0.6	0.8	1.0
						Blank control	-1.51	-0.60	0.41	-0.28	-0.82
Example one + H2O2	7.66	8.41	10.76	14.22	16.73
						Example one + H2O2	7.18	7.88	10.92	13.96	17.53
Example one + H2O2	6.65	8.20	11.45	12.78	15.50
						Example one + H2O2	6.17	7.77	10.49	15.50	16.78
Example one + H2O2	7.40	8.36	10.22	13.58	16.09
						Blank liposomes + H2O2	15.77	14.70	15.18	15.24	13.16
+H2O2	5.42	4.73	3.82	3.61	3.50
						+ complete culture medium	103.61	106.17	99.66	109.26	100.78

As can be seen from tables 5-2c, the cell viability was highest at 1.0mg/mL in example one, and H₂O₂Compared with the group (as shown in figure 10), the drug has certain protection effect on cells after being added.

TABLE 5-2d Hydrogen peroxide solution (1mmol/L) stimulation BV2 cell viability assay results (example two)

Concentration (mg/mL)	1.0	0.8	0.6	0.4	0.2
						Blank control	2.33	-0.66	1.37	0.52	0.62
Example two + H2O2	37.16	30.70	21.21	15.18	7.82
						Example two + H2O2	36.04	29.85	22.33	14.65	8.36
Example two + H2O2	35.24	31.29	21.26	12.62	6.49
						Example two + H2O2	36.66	30.92	22.12	15.34	7.93
Example two + H2O2	33.90	31.56	23.66	13.42	7.82
						Blank liposomes + H2O2	14.86	12.73	13.16	13.26	12.62
+H2O2	5.42	4.73	3.82	3.61	3.50
						+ complete culture medium	103.61	106.17	99.66	109.26	100.78

As can be seen from tables 5-2d, the cell viability was highest at 1.0mg/mL for example two, and H₂O₂Compared with the group (as shown in figure 10), the drug has certain protection effect on cells after being added.

TABLE 5-3a experimental results of blue light stimulated BV2 cell viability (example one)

Concentration (mg/mL)	0.2	0.4	0.6	0.8	1.0
						Blank control	-1.34	-0.19	-0.41	0.10	0.38
Example I + blue light	11.58	14.81	19.26	24.29	28.81
						Example I + blue light	11.72	15.46	18.54	24.29	29.53
Example I + blue light	15.74	17.90	18.76	23.50	28.30
						Example I + blue light	11.01	15.10	17.54	22.85	26.94
Example I + blue light	10.36	14.31	16.75	18.90	25.51
						Example I + blue light	10.79	15.46	18.11	24.21	28.52
Illuminate blue light	31.46	36.34	38.71	39.93	42.16
						+ complete culture medium	100.08	101.88	92.26	110.49	113.00

As can be seen from tables 5-3a, the survival rate of the cells is the highest when the first example is 1.0mg/mL, and the cells are protected after being added with the drug, compared with the group irradiated with blue light (see figure 13).

Tables 5-3b blue light stimulation BV2 cell viability assay results (example two)

As can be seen from tables 5-3b, the survival rate of the cells is the highest when the second example is 1.0mg/mL, and compared with the group irradiated with blue light (see FIG. 13), the cells are protected after the drug is added.

In conclusion, the second example has better protection effect than the first example, and not only can maintain the normal proliferation of BV2 cells, but also can maintain the normal activity of the cells.

Then, the effect of improving eyesight of example four was evaluated, and the results of the test using the MTT method (blue light irradiation for 15min) were shown in table 6.

TABLE 6 blue light stimulation BV2 cell viability assay results (example four)

	1	2	3	Mean value of
					Blank medium	0.92	0.31	0.68	0.64
Illuminate the blue light group	64.10	64.69	64.34	64.37
					Cassia seed	30.46	30.87	31.75	31.02
Honeysuckle	44.64	49.10	46.63	46.79
					Water chestnut	75.18	70.02	70.49	71.90
All-grass of Equisetum	14.81	11.94	13.46	13.40
					Cortex Fraxini	63.51	56.48	63.87	61.29
Flos Buddlejae	70.55	73.30	67.73	70.53
					Selfheal	61.99	62.69	57.65	60.78
Collagen liposome	44.94	47.69	44.64	45.76
					Vitamin C	89.07	86.57	88.83	88.16
Cassia seed liposome	24.36	38.66	41.36	34.80
					Licorice root, radix Glycyrrhizae	78.75	75.18	78.17	77.37
Flos Eriocauli	13.23	14.57	13.87	13.89
					Red matrimony vine	78.40	75.59	79.05	77.68
Glossy privet fruit	51.79	56.19	57.07	55.02
					Mint	76.76	69.43	77.23	74.47
Dendrobium nobile	76.35	81.74	80.98	79.69
					Hyaluronic acid	140.06	111.99	117.85	123.30
Gallic acid	80.41	76.28	74.96	77.22

As can be seen from table 6, the protective effect (from large to small) of the herbal extract on blue light stimulation of BV2 cells:

dendrobe, lycium ruthenicum, liquorice, mint and water chestnut

The twelve examples were prepared by selecting 5 kinds of the most effective herb extracts according to the above results, and the results of the test using the MTT method (blue light irradiation for 15min) were shown in Table 7, compared with the three examples.

TABLE 7 blue light stimulation BV2 cell viability assay results (example twelve and example three)

As can be seen from Table 7, the blue light-resisting effect of example three was superior to that of example twelve, and it can be concluded that the example three liposome used in the present invention was the most preferable.

Test example six products sensory and Effect test

Two types of common eye creams are sold in the market as a control group by taking the eleventh example as an experimental group. Volunteers (25-55 years old) male and female 15 persons each were selected, and were randomly and equally divided into 3 groups, and the experimental group and the control group were applied to the face after cleaning the skin every day, and the effect was observed after 15 days. The different items were scored according to the change in skin condition (0-100 points), and the results are shown in Table 8 below.

TABLE 8 Scoring results for different items

	EXAMPLE eleven	Control group one	Control group two
				Permeability of	85.67±3.06	81.83±1.61	87.67±2.52
Moisture retention	95.17±2.36	87.17±2.84	83.67±2.08
				Refreshing property	96.33±1.53	81.00±2.65	89.33±3.06
Wrinkle changes	90.67±3.21	81.33±1.53	89.67±1.53
				Skin color change	87.67±3.06	90.67±3.06	82.67±2.52
Change of edema	93.17±1.26	85.83±1.76	82.33±3.51
				Improvement of asthenopia	92.17±0.76	Is free of	Is free of

The results are combined to discover that the cassia seed compound collagen liposome prepared by the invention (example eleven) has the optimal effect, and particularly has the function of relieving the asthenopia which other eye cream products do not have.

The above embodiments are examples of the present invention, and the specific parameters in the above embodiments and examples are only for clearly showing the verification process of the present invention, and are not intended to limit the scope of the present invention. The protection scope of the present invention is defined by the claims, and all structural changes that can be made by using the contents of the description of the present invention are also included in the protection scope of the present invention.