CN117304062A

CN117304062A - Vegetable oil ceramide and its synthesis method and use

Info

Publication number: CN117304062A
Application number: CN202310186309.3A
Authority: CN
Inventors: 杨超文; 叶柳; 李青
Original assignee: Shenzhen Dikeman Biotechnology Co ltd
Current assignee: Shenzhen Dikeman Biotechnology Co ltd
Priority date: 2023-03-01
Filing date: 2023-03-01
Publication date: 2023-12-29

Abstract

The invention belongs to the technical field of biological medicines, and discloses vegetable oil ceramide, which is prepared by reacting vegetable oil fatty acid with sphingosine compounds, wherein the sphingosine compounds are selected from sphingosine, phytosphingosine and sphinganine, and the vegetable oil fatty acid is selected from sesame oil fatty acid, corn oil fatty acid, almond oil fatty acid, garlic oil fatty acid, peanut oil fatty acid, blackcurrant seed oil fatty acid, rapeseed oil fatty acid, hazelnut oil fatty acid, pumpkin seed oil fatty acid, DHA algae oil fatty acid, crab apple oil fatty acid, kiwi seed oil fatty acid, malva oil fatty acid and European Li Ziyou fatty acid. The scheme also discloses a synthesis method and application of the vegetable oil ceramide. The vegetable oil ceramide has excellent performances in the aspects of repairing natural skin barriers, anti-inflammatory, tissue healing, anti-aging and the like, and has wide application prospects in the fields of cosmetics, health-care products, biological medicines and the like.

Description

Vegetable oil ceramide and its synthesis method and use

Technical Field

The invention belongs to the technical field of biological medicine, and in particular relates to vegetable oil ceramide and a synthesis method and application thereof.

Background

Ceramides (ceramides, also known as molecular nails) naturally occur in the skin and are very important components of the skin barrier (stratum corneum), in amounts of up to 40-50 wt.%, ceramides are a class of sphingolipids consisting of long-chain bases of sphingosine and fatty acids, in which the carbon chain length, unsaturation and number of hydroxyl groups of the sphingosine moiety, fatty acid moiety are all variable, and ceramides represent a class of compounds. Ceramide has excellent properties in regulating skin barrier function, recovering skin moisture, enhancing adhesion between skin keratinocytes, and the like.

Because of the importance of ceramides, many cosmetic and pharmaceutical companies are researching and developing corresponding products. The natural plant-derived ceramide can form an effective skin barrier to prevent water loss and resist external damage due to the more sustainable and more environment-friendly raw material source and the characteristics similar to the skin ceramide components, and can become a next-generation environment-friendly, safe and reliable ceramide product.

The vegetable oil is prepared from fruits, seeds, germs, etc. of fat-rich plants by pretreatment such as cleaning, removing impurities, shelling, crushing, softening, rolling, extrusion puffing, etc., mechanical squeezing or solvent leaching to obtain crude oil, and refining. Fatty acids in vegetable oil can moisten and shiny skin. The content and the type of fatty acid of different vegetable oils are different, and other physiologically active substances are often contained.

Disclosure of Invention

The invention aims to provide ceramide synthesized by vegetable oil fatty acid.

Another object of the present invention is to provide a method for synthesizing vegetable oil ceramide, which uses vegetable oil grease or vegetable oil fatty acid which is natural and easily obtained as raw material.

It is another object of the present invention to provide the use of vegetable oil ceramides.

In order to achieve one of the above purposes, the present invention adopts the following technical scheme:

in a first aspect of the invention, a vegetable oil ceramide is obtained by reacting a vegetable oil fatty acid with a sphingosine compound selected from the group consisting of sphingosine, phytosphingosine, sphinganine, and a vegetable oil fatty acid selected from the group consisting of sesame oil fatty acid, corn oil fatty acid, almond oil fatty acid, garlic oil fatty acid, peanut oil fatty acid, blackcurrant seed oil fatty acid, rapeseed oil fatty acid, hazelnut oil fatty acid, pumpkin seed oil fatty acid, DHA algae oil fatty acid, begonia fruit oil fatty acid, kiwi seed oil fatty acid, marlura oil fatty acid, and european Li Ziyou fatty acid.

The reaction can be chemical synthesis reaction (as detailed below), or microbial fermentation method, i.e. using Pichia pastoris or Saccharomyces cerevisiae, fermenting under certain environment to obtain sphingosine compound, and adding fatty acid to obtain ceramide; or vegetable oil is used as a raw material, and proper bacterial strain is selected for fermentation to obtain the vegetable oil ceramide.

Sphingosine refers to 2-amino-4-octadecene-1, 3-diol, phytosphingosine refers to 2-amino-octadecane-1, 3, 4-triol, and dihydrosphingosine refers to 2-amino-octadecane-1, 3-diol.

Further, the vegetable oil fatty acid is obtained by hydrolyzing vegetable oil grease.

The sesame oil fatty acid comprises the following components: 45-70 wt% of linoleic acid, 15-35 wt% of oleic acid, 6-20 wt% of palmitic acid, 0-2 wt% of stearic acid, 0-1 wt% of arachidic acid and 0-0.5 wt% of linolenic acid.

Sesame oil is prepared by refining sesame after solvent extraction or squeezing after baking, and besides fatty acid, the sesame oil contains lignan compounds, the total content of which is up to 1.5-3.0 wt%, and the sesame oil is easy to hydrolyze and convert into pinoresinol under acidic conditions, so that the sesame oil has stronger oxidation resistance. Vitamin A, vitamin B, vitamin E, etc. in sesame oil can prevent fat from generating harmful substances damaging cells, remove toxin in vivo, effectively prevent various color spots such as cyasma, senile plaque, etc., and make skin show healthy luster.

The composition of the corn oil fatty acid is as follows: 50 to 70 weight percent of linoleic acid, 20 to 35 weight percent of oleic acid, 5 to 18 weight percent of palmitic acid, 0.5 to 3 weight percent of stearic acid, 0.01 to 1 weight percent of linolenic acid and 0 to 1 weight percent of arachidic acid.

Corn oil, also called maize oil, corn germ oil, is an oil extracted from corn germ. The content of unsaturated fatty acid in the corn oil is up to 80-85 wt%, and the corn oil is also rich in multiple vitamins and minerals. The corn oil is clear and transparent, has delicate fragrance and rich nutrition, can remove oxygen free radicals in the body, and has the effects of enhancing organism resistance, improving blood circulation, resisting oxidation and aging, promoting elastin and the like. In the traditional medical health care field, the corn oil can prevent atherosclerosis, muscular dystrophy, heart disease, reproductive dysfunction and thrombophlebitis, and obviously remove toxic effects of peroxide in human body on cell membranes, thereby improving oxygen utilization efficiency of muscles, nerves and tissues of the human body.

The composition of the almond oil fatty acid is as follows: 55 to 80 weight percent of oleic acid, 15 to 35 weight percent of linoleic acid, 0.1 to 8 weight percent of palmitic acid, 0.5 to 2 weight percent of stearic acid and 0.01 to 1 weight percent of linolenic acid.

The almond oil is excellent edible oil extracted from the almond, has various fatty acids required by human bodies, is rich in protein, vitamins, inorganic salts, dietary fibers and trace elements required by human bodies, can effectively relieve the skin itching phenomenon, and eliminates redness, dryness and inflammation; has the effects of moisturizing, anti-aging, antioxidation and the like, and is extremely mild. In the field of traditional Chinese medicine, the almond oil also has the effects of reducing blood fat and treating impetigo. The almond oil has wide application in cosmetics at home and abroad, and various brands of cream, milk honey and perfumed soap all use the almond oil as a natural additive. It not only increases the effects of moisturizing skin and resisting aging for the product, but also increases the warm fragrance for the product, and is a vegetable oil with excellent skin care and moisturizing effects.

The composition of the garlic oil fatty acid is as follows: 40 to 60 weight percent of nervonic acid, 30 to 50 weight percent of oleic acid, 5 to 12 weight percent of arachidic acid, 0 to 2 weight percent of palmitic acid and 0 to 2 weight percent of linoleic acid.

The oleum Bulbus Allii is a vegetable oil extracted from Bulbus Allii. Garlic is a unique single woody plant in China, is the plant with the highest content of nervonic acid, the oil content of the fruit is about 60wt%, and the proportion of nervonic acid is up to 40-60 wt%. Nervonic acid is an ultra-long chain monounsaturated fatty acid, which is a compound recognized by world scientists to repair and dredge nerve fibers of damaged cerebral nerve pathways and promote nerve cell regeneration. The research shows that the nervonic acid is the core natural component of animal brain nerve fiber and nerve cell and is essential nutrient for brain development and maintenance. Nervonic acid is a clinically indispensable pharmaceutical component for repairing after brain injury, and is also an essential component for development and regeneration of brain nerve fibers. The garlic oil is orange or brown, has high nutritive value, and has the effects of resisting oxidation, aging, bacteria, inflammation and the like when being applied to cosmetics. In traditional medicine, the garlic oil can restore and even enhance the activity of nerve endings, promote the growth and development of nerve cell tissues of organisms, and has good curative effects on patients suffering from cardiovascular and cerebrovascular diseases and autoimmune deficiency diseases of human bodies.

The peanut oil fatty acid comprises the following components: 40 to 60 weight percent of linoleic acid, 30 to 50 weight percent of oleic acid, 4 to 12 weight percent of palmitic acid, 0.5 to 5 weight percent of stearic acid, 0 to 1 weight percent of arachidic acid and 0 to 1 weight percent of behenic acid.

Peanut oil is a vegetable oil extracted from peanuts. Peanut oil is rich in various unsaturated fatty acids, resveratrol and various vitamins. Research shows that the peanut oil contains a great amount of nutrient substances which make the peanut oil have the functions of resisting bacteria, inflammation, allergy, thrombus, oxidation and free radicals; peanut oil diets can significantly reduce the total cholesterol level and low density value protein cholesterol levels of the body. In the traditional medical field, peanut oil has a prevention and treatment effect on type II diabetes mellitus and has a certain treatment effect on adhesive small intestinal obstruction.

The composition of the blackcurrant seed oil fatty acid is: 40 to 60 weight percent of linoleic acid, 20 to 40 weight percent of linolenic acid, 10 to 20 weight percent of oleic acid, 1 to 8 weight percent of palmitic acid, 0 to 2 weight percent of stearic acid and 0 to 2 weight percent of arachidic acid.

Blackcurrant seed oil, also called blackcurrant seed oil, is light yellow and has light fruit flavor. The blackcurrant seed oil has high nutritive value and contains a large amount of healthy fatty acid, wherein the gamma-linolenic acid can relieve climacteric discomfort, promote blood circulation, reduce fat retention in the inner wall of blood vessels, prevent and treat arteriosclerosis, reduce hypertension, maintain the health of cell membranes, ensure that cells retain more moisture, and enable skin to be smooth and elastic after long-term use. Blackcurrant seed oil can assist diabetics in controlling some complications. The blackcurrant seed oil has good effect of smearing the skin, is suitable for all the skins, can care and relieve the skin and hair, promote the updating of skin cells, delay the aging of the skin, and is very suitable for repairing and nursing damaged and sensitive skin. The black currant seed oil is used for adding soap, so that the effects of resisting bacteria, diminishing inflammation and resisting oxidation of the soap body can be improved, skin can be deeply nourished, skin allergy can be relieved, and the absorption of the skin to nutrient components can be accelerated.

The composition of the rapeseed oil fatty acid is as follows: 50 to 80 weight percent of oleic acid, 10 to 30 weight percent of linoleic acid, 2 to 8 weight percent of linolenic acid, 1 to 5 weight percent of stearic acid, 0.5 to 5 weight percent of palmitic acid, 0 to 1 weight percent of arachidic acid, 0 to 1 weight percent of arachidonic acid and 0 to 1 weight percent of erucic acid.

Rapeseed oil is an edible oil extracted from rapeseed. Rapeseed oil is rich in various fatty acids required by human body, and also contains rich tocopherol and brassicasterol, and has the effects of bacteriostasis, anti-inflammatory and the like. The canola oil can be treated to provide canola oil having a erucic acid content of no more than 3% of the fatty acid composition.

The hazelnut oil fatty acid comprises the following components: 70 to 90 weight percent of oleic acid, 8 to 25 weight percent of linoleic acid, 0.5 to 5 weight percent of palmitic acid, 0.5 to 4 weight percent of stearic acid, 0 to 0.5 weight percent of linolenic acid and 0 to 0.5 weight percent of palmitoleic acid.

Hazelnut oil is pale yellow and has a fine texture. The fatty acid component is mainly oleic acid and linoleic acid, and in addition, the nut-type nut-milk beverage also contains 8 amino acids required by human bodies, the amount of the fatty acid component is far higher than that of walnut, and the contents of various trace elements such as calcium, phosphorus and iron are also higher than that of other nuts. The hazelnut oil has excellent moisture retention capacity, can rapidly permeate the skin, and prevents water loss. The hazelnut oil contains a large amount of vitamin E and plant sterol, has a very good antioxidation effect, can prevent sunburn erythema spots, and can be matched with other sun-proof products for use. The hazelnut oil contains high-value protein, carbohydrate and vitamins, has good anti-inflammatory effect, is beneficial to skin regeneration and effectively prevents skin aging. The hazelnut oil is ideal oil for skin which is easy to grow acnes, has the effects of astringing and purifying the skin, is suitable for various skin types, can moisten and care dry sensitive skin, and can slow down the oil outlet speed of the skin due to the slight astringing effect.

The pumpkin seed oil fatty acid comprises the following components: 40 to 70 weight percent of linoleic acid, 20 to 55 weight percent of oleic acid, 3 to 10 weight percent of palmitic acid, 1 to 5 weight percent of stearic acid, 0.1 to 5 weight percent of linolenic acid, 0 to 1 weight percent of arachic acid and 0 to 0.5 weight percent of palmitoleic acid.

Pumpkin seed oil is called white melon seed oil and is rose red, contains various fatty acids, and has unsaturated fatty acid content of 90wt% and oleic acid and linoleic acid as main components; it also contains various bioactive substances such as phytosterol, amino acid, vitamins, carotenoid, minerals, etc., especially zinc, magnesium, calcium, and phosphorus. Linoleic acid and linolenic acid are essential fatty acids for the human body, and are necessary for the synthesis of phospholipids, the formation of cellular structures, and the maintenance of normal functions of tissues. Therefore, the pumpkin seed oil is a good nutritional and health-care oil.

The DHA algae oil fatty acid comprises the following components: 50 to 75 weight percent of DHA, 15 to 35 weight percent of oleic acid, 3 to 15 weight percent of palmitic acid, 0.1 to 1 weight percent of stearic acid, 0.1 to 1 weight percent of EPA, 0.01 to 0.5 weight percent of linolenic acid, 0 to 0.5 weight percent of arachidic acid and 0 to 0.5 weight percent of behenic acid.

DHA, docosahexaenoic acid, commonly known as brain gold, is a polyunsaturated fatty acid very important for human body, and belongs to an important member of Omega-3 unsaturated fatty acid family. DHA is a main element for growth and maintenance of nervous system cells, is an important fatty acid for forming brain and retina, has content of up to 20wt% in human brain cortex, and has maximum content in retina of about 50wt%, and is important for infant intelligence and vision development. DHA algae oil is extracted from marine microalgae, is not transmitted by food chains, is relatively safer, and has high DHA content. DHA extracted from algae is purely natural, plant, strong in antioxidant capacity and low in EPA content, is most beneficial to infant absorption, and can effectively promote the development of retina and brain; DHA extracted from deep sea fish oil has active property, is easy to oxidize and denature, and has high EPA content.

The composition of the crab apple oil fatty acid is as follows: 30 to 50 weight percent of oleic acid, 15 to 40 weight percent of linoleic acid, 8 to 20 weight percent of stearic acid, 6 to 20 weight percent of palmitic acid, 0.2 to 1.5 weight percent of arachidic acid, 0.05 to 2 weight percent of linolenic acid, 0.05 to 1 weight percent of palmitoleic acid, 0 to 1 weight percent of behenic acid and 0 to 0.5 weight percent of arachidonic acid.

The crab apple oil is also called as inophen oil, is a vegetable oil with extremely high medicinal value, contains various essential oil components such as thymol, terpene compounds and aromatic acid, and has the anti-tumor potential by unique components of pyranocoumarin derivatives, wherein the pyranocoumarin can inhibit elastase, prevent wrinkles and skin from loosening, recover the elasticity of the skin and lighten facial lines. The crab oil has good skin-friendly property and high antioxidant activity, is very suitable for being used as base oil of cosmetic products, and can reduce the oxidation speed; also has antiinflammatory and skin caring effects.

The composition of the kiwi seed oil fatty acid is as follows: 45-70 wt% of linolenic acid, 8-20 wt% of linoleic acid, 5-20 wt% of oleic acid, 2-10 wt% of palmitic acid and 0.5-5 wt% of stearic acid.

The kiwi seed oil (kiwi seed oil) is pure natural vegetable oil mainly containing essential fatty acid linolenic acid and linoleic acid, is rich in active substances such as vitamin E, trace element Se and the like, has the effects of moisturizing, whitening and removing freckles, and can be applied to skin-cleaning, skin-protecting and skin-nourishing cosmetics.

The composition of the brucella oil fatty acid is as follows: 65 to 85 weight percent of oleic acid, 5 to 15 weight percent of palmitic acid, 2 to 10 weight percent of linoleic acid, 2 to 10 weight percent of stearic acid, 0.2 to 2 weight percent of arachidic acid, 0 to 1 weight percent of arachidonic acid and 0 to 0.5 weight percent of linolenic acid.

The Malala oil is a clear, pale yellow liquid with a nutty aroma. The main fatty acids of the Malla oil are oleic acid and linoleic acid, and are also rich in very strong antioxidants, which makes the oiliness of the Malla oil extremely stable and not easily oxidized.

The composition of the european Li Ziyou fatty acid is: 60 to 80 weight percent of oleic acid, 10 to 30 weight percent of linoleic acid, 0.5 to 5 weight percent of stearic acid, 0.5 to 3 weight percent of palmitoleic acid, 0 to 7 weight percent of palmitic acid and 0 to 1 weight percent of linolenic acid.

Vegetable oil fatty acids are affected by plant species, soil, climate, production area, picking season, and extraction process, and the content of each component varies.

Vegetable oil ceramide, its composition includes: oleic acid ceramide, linoleic acid ceramide, palmitic acid ceramide.

Specifically, the composition of the vegetable oil ceramide is: 10 to 90 weight percent of oleic acid ceramide, 2 to 70 weight percent of linoleic acid ceramide and 0.1 to 20 weight percent of palmitic acid ceramide.

The vegetable oil ceramide further comprises at least one of linolenic acid ceramide, DHA ceramide, nervonic acid ceramide, arachidic acid ceramide, stearic acid ceramide, behenic acid ceramide, arachidonic acid ceramide, erucic acid ceramide, palmitoleic acid ceramide and EPA ceramide.

Specifically, the vegetable oil ceramides further comprise at least one of no more than 75wt% DHA ceramides, no more than 60wt% nervonic acid ceramides, no more than 70wt% linolenic acid ceramides, no more than 12wt% arachidic acid ceramides, no more than 20wt% stearic acid ceramides, no more than 1wt% behenic acid ceramides, no more than 1wt% arachidonic acid ceramides, no more than 1wt% erucic acid ceramides, no more than 1wt% palmitoleic acid ceramides, and no more than 1wt% EPA ceramides.

In particular 0.1 to 75 wt.% of DHA ceramide, 0.1 to 60 wt.% of nervonic acid ceramide, 0.1 to 70 wt.% of linolenic acid ceramide, 0.1 to 12 wt.% of arachidic acid ceramide, 0.1 to 20 wt.% of stearic acid ceramide, 0.1 to 1 wt.% of behenic acid ceramide, 0.1 to 1 wt.% of arachidonic acid ceramide, 0.1 to 1 wt.% of erucic acid ceramide, 0.1 to 1 wt.% of palmitoleic acid ceramide, 0.1 to 1 wt.% of EPA ceramide.

Specifically, the vegetable oil ceramide also comprises 0 to 75 weight percent of DHA ceramide, 0 to 60 weight percent of nervonic acid ceramide, 0 to 70 weight percent of linolenic acid ceramide, 0 to 12 weight percent of arachidic acid ceramide, 0 to 20 weight percent of stearic acid ceramide, 0 to 1 weight percent of behenic acid ceramide, 0 to 1 weight percent of arachidonic acid ceramide, 0 to 1 weight percent of erucic acid ceramide, 0 to 1 weight percent of palmitoleic acid ceramide and 0 to 1 weight percent of EPA ceramide.

The vegetable oil ceramide also comprises vitamin A, vitamin B, vitamin E, carotenoid, resveratrol, tocopherol, thymol, terpene compounds, aromatic acid, dipyranocoumarin, sterol, amino acid, trace elements and the like which exist in vegetable oil fatty acid but do not react with sphingosine compounds.

Oleic acid ceramide is obtained by condensation reaction of oleic acid and sphingosine compounds, and comprises oleic acid phytosphingosine ceramide, oleic acid sphingosine ceramide and oleic acid dihydrosphingosine ceramide; the linoleic acid ceramide is obtained by condensation reaction of linoleic acid and sphingosine compounds, and comprises linoleic acid phytosphingosine ceramide, linoleic acid sphingosine ceramide and linoleic acid dihydrosphingosine ceramide; the palmitic acid ceramide is obtained by condensation reaction of palmitic acid and sphingosine compounds, and comprises palmitic acid phytosphingosine ceramide, palmitic acid sphingosine ceramide and palmitic acid dihydrosphingosine ceramide; DHA ceramide, nervonic acid ceramide, linolenic acid ceramide, arachidic acid ceramide, stearic acid ceramide, behenic acid ceramide, arachidonic acid ceramide, erucic acid ceramide, palmitoleic acid ceramide, EPA ceramide, and the like, and so on.

In a second aspect of the invention, a method for synthesizing vegetable oil ceramide comprises the following steps:

under the conditions of condensing agent and organic base, vegetable oil fatty acid reacts with sphingosine compound, the condensing agent is DCC, and the organic base is DMAP.

Further, the molar ratio of the vegetable oil fatty acid to the sphingosine compound to the DCC to the DMAP is 1: (1-1.5): (1-2): (0.2-0.5), wherein the solvent for the reaction is at least one of dichloromethane, tetrahydrofuran, ethyl acetate and acetonitrile.

Vegetable oils purchased on the market are generally in the form of oils and fats, which need to be hydrolysed to vegetable oil fatty acids, and therefore comprise the following steps:

the vegetable oil fat is hydrolyzed by saponification reaction to obtain vegetable oil fatty acid.

Further, the saponification reaction is hydrolysis of vegetable oil grease in potassium hydroxide solution.

Further, the mass ratio of the vegetable oil and fat to the potassium hydroxide is 1: (1-2).

In a third aspect of the invention, the use of a vegetable oil ceramide in a cosmetic, pharmaceutical, dietary or health product.

Further, the vegetable oil ceramide has at least one of skin barrier repair, tissue healing, anti-aging, anti-inflammatory, anti-photoaging, antioxidant, collagen synthesis promoting, elastin activity maintaining, and whitening effects.

A composition comprising a vegetable oil ceramide, said composition having at least one of skin barrier repair, tissue healing, anti-aging, anti-inflammatory, anti-photoaging, antioxidant, promoting collagen synthesis, maintaining elastin viability, whitening efficacy.

The composition contains acceptable auxiliary materials, including one or more of solubilizer, antiseptic, antioxidant, pH regulator, penetration enhancer, liposome, humectant, thickener, chelating agent, skin feel regulator, surfactant, emulsifier, essence and pigment; the composition is in the form of cream, emulsion, solution, film, aerosol or spray.

The invention has the following beneficial effects:

the fatty acid of the vegetable oil and the sphingosine compound naturally existing in the skin are subjected to mild reaction to prepare the vegetable oil ceramide, which has excellent performance in the aspects of repairing, resisting oxidation, resisting aging and the like of the natural barrier of the skin, and has wide application prospect in the fields of cosmetics, health care products, biological medicines and the like.

1. Better results compared to ceramide alone. Different ceramides have different effects due to the structural differences, and ceramides with a single structure generally have difficulty in having comprehensive effects. The scheme is based on a bionic thought, and natural vegetable oil grease or fatty acid is used as a raw material to synthesize the compound ceramide so as to make up the difference of different ceramide effects, and trace fatty acid in vegetable oil can form trace ceramide to play a role in efficacy supplement.

2. Compared with the compounded ceramide, the effect is better. Besides fatty acid (or grease), the vegetable oil also contains vitamin A, carotenoid, resveratrol, tocopherol, sterol, amino acid, trace elements and other components, and the nutrient substances have different physiological activities, and the ceramide synthesized by the vegetable oil has synergistic effect with other active components contained in the vegetable oil, so that the ceramide compounded according to similar proportion has better effect.

3. The cost is lower. The method of the invention can rapidly obtain the composition compounded by various ceramides, the vegetable oil and fat or the fatty acid thereof has wide sources, is easy to obtain commercially, has lower cost, is more environment-friendly and economical, is different from the idea of mixing and compounding different single ceramides, has high raw material price, needs to separately produce different ceramides, and then is compounded, thereby increasing the preparation cost.

4. The synthesis method is simple. The method can adopt chemical synthesis to realize one-step preparation of various ceramides, and can also use a microbial fermentation method.

Drawings

FIG. 1 is a bar graph showing the results of the cell proliferation activity test of example 15;

FIGS. 2, 3 and 4 show the results of the cell migration ability test of example 16;

FIGS. 5, 6, 7 and 8 are bar graphs of elastase inhibition ratios of example 17;

FIG. 9 is a bar graph showing the detection of IL-6 factor expression level in anti-inflammatory repair efficacy in example 18;

FIGS. 10, 11, 12, 13, 14 and 15 are bar graphs showing MMP1 expression levels in the photo-aging test of example 19;

FIGS. 16, 17, 18, 19 are bar graphs of DPPH radical scavenging for oxidation resistance test of example 20;

figures 20, 21 and 22 are bar graphs showing the whitening activity test melanin content of example 21.

Detailed Description

The invention will be further illustrated with reference to specific examples. DCC means N, N' -dicyclohexylcarbodiimide and DMAP means 4-dimethylaminopyridine. The silica gel column chromatography uses Qingdao ocean silica gel (particle size 0.040-0.063 mm). Thin Layer Chromatography (TLC) using 60F254 silica gel plates was performed using UV light (254 nm) or iodine.

Example 1

Synthesis of ceramide from sesame oil fatty acid and phytosphingosine

The first step: 50g of sesame oil and fat are dissolved in 60mL of tetrahydrofuran, cooled in an ice bath, 110mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the completion of the dropwise addition until the TLC detection reaction is completed.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 120mL of ethyl acetate to extract the water phase, and adding 80mL of saturated saline Washing once, organic phase added anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give 42g of sesame oil fatty acid.

And a second step of: sesame oil fatty acid (50 mmol, based on the main component fatty acid), DCC (70 mmol), DMAP (10 mmol) were added to a 250mL round bottom flask, and 80mL dichloromethane was added, followed by stirring at room temperature for 1 hour, followed by phytosphingosine (75 mmol) being added to the reaction system, and stirring at room temperature until TLC detection was completed.

Post-treatment: adding water for quenching reaction, separating organic layer, drying, filtering, vacuum concentrating, washing with solvent to obtain sesame oil ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3 dplus) with Innoval ODS-2.6x250 mm,5 μm column temperature: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linoleic acid-phytosphingosine ceramide 9.5min, palmitic acid-phytosphingosine ceramide 10.7min, oleic acid-phytosphingosine ceramide 11.3min, stearic acid-phytosphingosine ceramide 13.9min.

The obtained product is analyzed by high performance liquid chromatography, the contents of linoleic acid-phytosphingosine ceramide, oleic acid-phytosphingosine ceramide, palmitic acid-phytosphingosine ceramide and stearic acid-phytosphingosine ceramide are 62%, 23%, 10%, 1% in sequence, and the rest is other components, and the content is less.

Example 2

Synthesis of ceramide from corn oil fatty acid and sphingosine

The first step: 50g of corn oil is dissolved in 60mL of tetrahydrofuran, cooled in an ice bath, 110mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the completion of the dropwise addition until TLC detection is completed.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 120mL of ethyl acetate to extract a water phase, adding 80mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Drying, filtration and concentration in vacuo gave 41.2g of corn oil fatty acid.

And a second step of: corn oil fatty acid (50 mmol, based on the main component fatty acid), DCC (60 mmol), DMAP (15 mmol) were added to a 250mL round bottom flask, followed by 100mL dichloromethane, followed by stirring at room temperature for 1 hour, followed by sphingosine (75 mmol) added to the reaction system, followed by stirring at room temperature until TLC detection was complete.

Post-treatment: adding water for quenching reaction, separating organic layer, drying, filtering and vacuum concentrating, washing with solvent to obtain corn oil ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3 dplus) with Innoval ODS-2.6x250 mm,5 μm column temperature: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linolenic acid-sphingosine ceramide 7.9min, linoleic acid-sphingosine ceramide 8.7min, oleic acid-sphingosine ceramide 10.2min, palmitic acid-sphingosine ceramide 10.4min, stearic acid-sphingosine ceramide 13.6min, arachidic acid-sphingosine ceramide 17.8min.

The obtained products are analyzed by high performance liquid chromatography, the contents of linoleic acid-sphingosine ceramide, oleic acid-sphingosine ceramide, palmitic acid-sphingosine ceramide, stearic acid-sphingosine ceramide, linolenic acid-sphingosine ceramide and arachidic acid-sphingosine ceramide are 56%, 29%, 9%, 1% and 1% in sequence, and the rest are other components, and the contents are less.

Example 3

Synthesis of ceramide from almond oil fatty acid and sphinganine

The first step: 50g of almond oil fat is dissolved in 60mL of tetrahydrofuran, cooled in an ice bath, 110mL of potassium hydroxide (25 wt%) solution is added dropwise, and the reaction is carried out after the dropwise addition is completed, and the temperature is raised to room temperature until the TLC detection reaction is completed.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 120mL of ethyl acetate to extract the water phase, and adding 80mL of saturated solutionWashing with salt water once, adding anhydrous Na into the organic phase ₂ SO ₄ Drying, filtration and concentration in vacuo gave 39.9g of almond oil fatty acid.

And a second step of: almond oil fatty acid (50 mmol, based on main component fatty acid), DCC (55 mmol), DMAP (25 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, then sphinganine (60 mmol) was added to the reaction system, and stirring at room temperature was performed until TLC detection was complete.

Post-treatment: adding water for quenching reaction, separating organic layer, drying, filtering, vacuum concentrating, washing with solvent to obtain almond oil ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3 dplus) with Innoval ODS-2.6x250 mm,5 μm column temperature: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linolenic acid-dihydrosphingosine ceramide 8.3min, linoleic acid-dihydrosphingosine ceramide 9.5min, palmitic acid-dihydrosphingosine ceramide 10.6min, oleic acid-dihydrosphingosine ceramide 11.1min, stearic acid-dihydrosphingosine ceramide 13.6min.

The obtained products are analyzed by high performance liquid chromatography, the content of oleic acid-dihydrosphingosine ceramide, linoleic acid-dihydrosphingosine ceramide, palmitic acid-dihydrosphingosine ceramide, stearic acid-dihydrosphingosine ceramide and linolenic acid-dihydrosphingosine ceramide is 62%, 24%, 7%, 2%, 1% in sequence, and the rest is other components, and the content is less.

Example 4

Synthesis of ceramide from oleum Bulbus Allii oil acid and phytosphingosine

The first step: 50g of garlic oil and fat are dissolved in 80mL of tetrahydrofuran, cooled in an ice bath, 130mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dripping is finished until TLC detection is finished.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, addingExtracting the aqueous phase with 140mL of ethyl acetate, adding 100mL of saturated saline, washing once with water, and adding the organic phase with anhydrous Na ₂ SO ₄ Drying, filtering and concentrating under vacuum to obtain 38.2g of oleum Bulbus Allii oil fatty acid.

And a second step of: garlic oil fatty acid (50 mmol, based on main component fatty acid), DCC (70 mmol) and DMAP (10 mmol) were added to a 250mL round bottom flask, 80mL dichloromethane was added, followed by stirring at room temperature for 1 hour, and phytosphingosine (70 mmol) was added to the reaction system, followed by stirring at room temperature until TLC detection was completed.

Post-treatment: adding water for quenching reaction, separating an organic layer, drying, filtering and concentrating in vacuum, washing by a solvent to obtain garlic fruit oil ceramide, and analyzing a product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linoleic acid-phytosphingosine ceramide 9.5min, palmitic acid-phytosphingosine ceramide 10.7min, oleic acid-phytosphingosine ceramide 11.3min, arachidic acid-phytosphingosine ceramide 16.5min, and ceramide 22.2min.

The obtained product is analyzed by high performance liquid chromatography, and the content of the nervonic acid-phytosphingosine ceramide, oleic acid-phytosphingosine ceramide, arachidic acid-phytosphingosine ceramide, palmitic acid-phytosphingosine ceramide and linoleic acid-phytosphingosine ceramide is 52%, 35%, 8%, 2%, 1% in sequence, and the rest is other components, and the content is less.

Example 5

Synthesis of ceramide from peanut oil fatty acid and sphingosine

The first step: 40g of peanut oil is dissolved in 50mL of tetrahydrofuran, cooled in an ice bath, 100mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the completion of the dropwise addition until the completion of TLC detection.

Post-treatment: adding thinHydrochloric acid (3N) was used to adjust the pH of the reaction system to 3, 100mL of ethyl acetate was added to extract the aqueous phase, 80mL of saturated brine was added to wash the aqueous phase once, and the organic phase was added to anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give 32.1g of peanut oil fatty acid.

And a second step of: peanut oil fatty acid (50 mmol, based on the main component fatty acid), DCC (65 mmol), DMAP (15 mmol) were added to a 250mL round bottom flask, followed by 100mL dichloromethane, followed by stirring at room temperature for 1 hour, followed by sphingosine (65 mmol) added to the reaction system, followed by stirring at room temperature until TLC detection was complete.

Post-treatment: adding water for quenching reaction, separating an organic layer, drying, filtering and concentrating in vacuum, washing by a solvent to obtain peanut oil ceramide, and analyzing a product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3 dplus) with Innoval ODS-2.6x250 mm,5 μm column temperature: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linoleic acid-sphingosine ceramide 8.7min, oleic acid-sphingosine ceramide 10.2min, palmitic acid-sphingosine ceramide 10.5min, stearic acid-sphingosine ceramide 13.6min, and behenic acid-sphingosine ceramide 24.0min.

The obtained product is analyzed by high performance liquid chromatography, the contents of linoleic acid-sphingosine ceramide, oleic acid-sphingosine ceramide, palmitic acid-sphingosine ceramide, stearic acid-sphingosine ceramide and behenic acid-sphingosine ceramide are 43%, 7%, 3% and 1% in sequence, and the rest are other components, and the content is less.

Example 6

Synthesis of ceramide from blackcurrant seed oil fatty acid and sphinganine

The first step: 50g of blackcurrant seed oil grease is dissolved in 80mL of tetrahydrofuran, cooled in an ice bath, 140mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dropwise addition until TLC detection is finished.

Post-treatment: adding dilute hydrochloric acid(3N) adjusting pH of the reaction system to 3, adding 120mL of ethyl acetate to extract the aqueous phase, adding 100mL of saturated saline water once, and adding the organic phase to anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to yield 43.1g of blackcurrant seed oil fatty acid.

And a second step of: blackcurrant seed oil fatty acid (50 mmol based on main component fatty acid), DCC (90 mmol), DMAP (20 mmol) were added to a 250mL round bottom flask, and 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, followed by addition of sphinganine (65 mmol) to the reaction system, stirring at room temperature, and detection by TLC was completed.

Post-treatment: adding water for quenching reaction, separating an organic layer, drying, filtering and concentrating in vacuum, washing by a solvent to obtain blackcurrant seed oil ceramide, and analyzing a product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linolenic acid-dihydrosphingosine ceramide 8.3min, linoleic acid-dihydrosphingosine ceramide 9.5min, palmitic acid-dihydrosphingosine ceramide 10.6min, oleic acid-dihydrosphingosine ceramide 11.1min, stearic acid-dihydrosphingosine ceramide 13.5min, arachidic acid-dihydrosphingosine ceramide 16.0min.

The obtained products are analyzed by high performance liquid chromatography, the contents of linoleic acid-dihydrosphingosine ceramide, linolenic acid-dihydrosphingosine ceramide, oleic acid-dihydrosphingosine ceramide, palmitic acid-dihydrosphingosine ceramide, stearic acid-dihydrosphingosine ceramide and arachidic acid-dihydrosphingosine ceramide are 45%, 31%, 13%, 5%, 2% and 2% in sequence, and the rest are other components, and the content is less.

Example 7

Synthesis of ceramide from rapeseed oil fatty acid and phytosphingosine

The first step: 50g of rapeseed oil is dissolved in 70mL of tetrahydrofuran, cooled in an ice bath, 120mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dropwise addition until TLC detection is completed.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 120mL of ethyl acetate to extract a water phase, adding 80mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Drying, filtration and concentration in vacuo gave 41.3g rapeseed oil fatty acid.

And a second step of: rapeseed oil fatty acid (50 mmol, calculated as main component fatty acid), DCC (80 mmol), DMAP (20 mmol) were added to a 250mL round bottom flask, followed by 80mL dichloromethane, followed by stirring at room temperature for 1 hour, followed by phytosphingosine (70 mmol) added to the reaction system, followed by stirring at room temperature until TLC detection was complete.

Post-treatment: adding water for quenching reaction, separating an organic layer, drying, filtering and concentrating in vacuum, washing with a solvent to obtain rapeseed oil ceramide, and analyzing a product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3 dplus) with Innoval ODS-2.6x250 mm,5 μm column temperature: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linolenic acid-phytosphingosine ceramide 8.3min, linoleic acid-phytosphingosine ceramide 9.5min, palmitic acid-phytosphingosine ceramide 10.7min, oleic acid-phytosphingosine ceramide 11.3min, stearic acid-phytosphingosine ceramide 13.9min, erucic acid-phytosphingosine ceramide 14.2min.

The obtained product is analyzed by high performance liquid chromatography, and the content of oleic acid-phytosphingosine ceramide, linoleic acid-phytosphingosine ceramide, linolenic acid-phytosphingosine ceramide, stearic acid-phytosphingosine ceramide, palmitic acid-phytosphingosine ceramide and erucic acid-phytosphingosine ceramide is 68%, 12%, 5.5%, 5%, 1% and the rest is other components, and the content is less.

Example 8

Synthesis of ceramide from European Li Ziyou fatty acids and sphingosine

The first step: 50g European Li Ziyou grease was dissolved in 100mL tetrahydrofuran, cooled in an ice bath, 150mL potassium hydroxide (25 wt%) solution was added dropwise, and after the addition was completed, the reaction was allowed to stand at room temperature until the TLC detection reaction was completed.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 120mL of ethyl acetate to extract a water phase, adding 100mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Dried, filtered and concentrated in vacuo to give 42.4g of European Li Ziyou fatty acid.

And a second step of: european Li Ziyou fatty acid (50 mmol based on main component fatty acid), DCC (75 mmol) and DMAP (15 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, and then sphingosine (70 mmol) was added to the reaction system, followed by stirring at room temperature until TLC detection was complete.

Post-treatment: adding water to quench the reaction, separating an organic layer, drying, filtering and concentrating in vacuum, washing by a solvent to obtain European Li Ziyou ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: palmitoleic acid-sphingosine ceramide 8.2min, linoleic acid-sphingosine ceramide 8.7min, oleic acid-sphingosine ceramide 10.1min, palmitic acid-sphingosine ceramide 10.4min, stearic acid-sphingosine ceramide 13.6min.

The obtained product is analyzed by high performance liquid chromatography, the content of oleic acid-sphingosine ceramide, linoleic acid-sphingosine ceramide, stearic acid-sphingosine ceramide, palmitoleic acid-sphingosine ceramide and palmitoleic acid-sphingosine ceramide is 73%, 10%, 4.5%, 3% and 7% in sequence, and the rest is other components, and the content is less.

Example 9

Ceramide synthesis by hazelnut oil fatty acid and sphinganine

The first step: 50g hazelnut oil is dissolved in 70mL tetrahydrofuran, cooled in ice bath, 120mL potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the addition until TLC detection is completed.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 150mL of ethyl acetate to extract a water phase, adding 100mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Drying, filtering and vacuum concentrating to obtain 37.9g hazelnut oil fatty acid.

And a second step of: hazelnut oil fatty acid (50 mmol, calculated as main component fatty acid), DCC (85 mmol), DMAP (25 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, then sphinganine (75 mmol) was added to the reaction system, and stirring at room temperature was performed until TLC detection was completed.

Post-treatment: filtering the reaction solution to remove solids, collecting filtrate, respectively adding a saturated solution of sodium bicarbonate, 1N diluted hydrochloric acid and saturated saline solution for washing, separating an organic layer, drying, filtering, concentrating in vacuum, washing with a solvent to obtain hazelnut oil ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linolenic acid-dihydrosphingosine ceramide 8.3min, linoleic acid-dihydrosphingosine ceramide 9.5min, palmitic acid-dihydrosphingosine ceramide 10.6min, oleic acid-dihydrosphingosine ceramide 11.2min, stearic acid-dihydrosphingosine ceramide 13.5min.

The obtained products are analyzed by high performance liquid chromatography, the content of oleic acid-dihydrosphingosine ceramide, linoleic acid-dihydrosphingosine ceramide, palmitic acid-dihydrosphingosine ceramide, stearic acid-dihydrosphingosine ceramide and linolenic acid-dihydrosphingosine ceramide is 72%, 18%, 4%, 2%, 0.5% in sequence, and the balance is other components, and the content is less.

Example 10

Synthesis of ceramide from pumpkin seed oil fatty acid and phytosphingosine

The first step: 50g of pumpkin seed oil grease is dissolved in 60mL of tetrahydrofuran, cooled in an ice bath, 100mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dropwise addition is finished until TLC detection is finished.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 140mL of ethyl acetate to extract a water phase, adding 100mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Drying, filtering and vacuum concentrating to obtain 42.2g pumpkin seed oil fatty acid.

And a second step of: pumpkin seed oil fatty acid (50 mmol, calculated as main component fatty acid), DCC (85 mmol), DMAP (20 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, and phytosphingosine (65 mmol) was added to the reaction system, followed by stirring at room temperature until TLC detection was completed.

Post-treatment: filtering the reaction solution to remove solids, collecting filtrate, respectively adding a saturated solution of sodium bicarbonate, washing with 1N diluted hydrochloric acid and saturated saline, separating an organic layer, drying, filtering, concentrating in vacuum, washing with a solvent to obtain pumpkin seed oil ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linolenic acid-phytosphingosine ceramide 8.3min, linoleic acid-phytosphingosine ceramide 9.5min, palmitic acid-phytosphingosine ceramide 10.7min, oleic acid-phytosphingosine ceramide 11.3min, stearic acid-phytosphingosine ceramide 13.9min, arachidic acid-phytosphingosine ceramide 16.4min.

The obtained product is analyzed by high performance liquid chromatography, the content of linoleic acid-phytosphingosine ceramide, oleic acid-phytosphingosine ceramide, palmitic acid-phytosphingosine ceramide, stearic acid-phytosphingosine ceramide, linolenic acid-phytosphingosine ceramide and arachidic acid-phytosphingosine ceramide is 60%, 22%, 7%, 4%, 2.5%, 1% and the rest is other components, and the content is less.

Example 11

DHA algae oil fatty acid and sphingosine synthesis ceramide

The first step: 50g of DHA algae oil is dissolved in 100mL of tetrahydrofuran, cooled in ice bath, 150mL of potassium hydroxide (25 wt%) solution is added dropwise, and the reaction is carried out after the dropwise addition is completed, and the temperature is raised to room temperature until the TLC detection reaction is completed.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 140mL of ethyl acetate to extract a water phase, adding 100mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Drying, filtering and concentrating in vacuum to obtain 39.9g DHA algae oil fatty acid.

And a second step of: DHA algae oil fatty acid (50 mmol, calculated as main component fatty acid), DCC (70 mmol) and DMAP (15 mmol) are added into a 250mL round bottom flask, 100mL dichloromethane is added, then stirring is carried out for 1 hour at room temperature, then sphingosine (60 mmol) is added into a reaction system, stirring is carried out at room temperature, and the reaction is detected by TLC.

Post-treatment: filtering the reaction solution to remove solids, collecting filtrate, respectively adding a saturated solution of sodium bicarbonate, washing with 1N diluted hydrochloric acid and saturated saline, separating an organic layer, drying, filtering, concentrating in vacuum, washing with a solvent to obtain DHA algae oil ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 2% water +98% methanol.

The retention time of each component HPLC was: linolenic acid-sphingosine ceramide 7.9min, oleic acid-sphingosine ceramide 10.2min, palmitic acid-sphingosine ceramide 10.5min, stearic acid-sphingosine ceramide 13.6min, EPA-sphingosine ceramide 15.6min, arachidic acid-sphingosine ceramide 17.8min, DHA-sphingosine ceramide 19.1min, and behenic acid-sphingosine ceramide 24.0min.

The obtained product is analyzed by high performance liquid chromatography, and the content of DHA-sphingosine ceramide, oleic acid-sphingosine ceramide, palmitic acid-sphingosine ceramide, stearic acid-sphingosine ceramide, linolenic acid-sphingosine ceramide, EPA-sphingosine ceramide, arachidic acid-sphingosine ceramide and behenic acid-sphingosine ceramide is 62%, 23%, 10%, 1%, 0.5% and 0.5% in sequence, and the balance is other components, and the content is less.

Example 12

Synthesis of ceramide from fatty acid and sphinganine of crab oil

The first step: 50g of crab apple oil grease is dissolved in 60mL of tetrahydrofuran, cooled in ice bath, 120mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dripping is finished until TLC detection is finished.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 120mL of ethyl acetate to extract a water phase, adding 120mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Drying, filtering and concentrating in vacuum to obtain 40.6g of crab apple oil fatty acid.

And a second step of: malus spectabilis oil fatty acid (50 mmol, calculated as main component fatty acid), DCC (60 mmol) and DMAP (20 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, and then sphinganine (60 mmol) was added to the reaction system, followed by stirring at room temperature until TLC detection was complete.

Post-treatment: filtering the reaction solution to remove solids, collecting filtrate, respectively adding a saturated solution of sodium bicarbonate, 1N diluted hydrochloric acid and saturated saline solution for washing, separating an organic layer, drying, filtering, concentrating in vacuum, washing with a solvent to obtain the malus spectabilis oil ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linolenic acid-dihydrosphingosine ceramide 8.3min, palmitoleic acid-dihydrosphingosine ceramide 8.9min, linoleic acid-dihydrosphingosine ceramide 9.5min, oleic acid-dihydrosphingosine ceramide 11.2min, palmitic acid-dihydrosphingosine ceramide 10.6min, stearic acid-dihydrosphingosine ceramide 13.5min, arachidic acid-dihydrosphingosine ceramide 16.0min.

The obtained products are analyzed by high performance liquid chromatography, and the content of oleic acid-dihydrosphingosine ceramide, linoleic acid-dihydrosphingosine ceramide, stearic acid-dihydrosphingosine ceramide, palmitic acid-dihydrosphingosine ceramide, arachidic acid-dihydrosphingosine ceramide, linolenic acid-dihydrosphingosine ceramide and palmitoleic acid-dihydrosphingosine ceramide is 47%, 22%, 13%, 11%, 1.5% and 1% in sequence, and the rest is other components, and the content is less.

Example 13

Synthesis of ceramide from kiwi seed oil fatty acid and phytosphingosine

The first step: 50g of kiwi fruit seed oil grease is dissolved in 60mL of tetrahydrofuran, cooled in an ice bath, 120mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dropwise addition until TLC detection is finished.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 140mL of ethyl acetate to extract a water phase, adding 100mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Drying, filtration and concentration in vacuo gave 39.5g of kiwi seed oil fatty acid.

And a second step of: kiwi seed oil fatty acid (50 mmol, calculated as main component fatty acid), DCC (70 mmol), DMAP (15 mmol) were added to a 250mL round bottom flask, and 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, and phytosphingosine (65 mmol) was added to the reaction system, followed by stirring at room temperature until TLC detection was completed.

Post-treatment: filtering the reaction solution to remove solids, collecting filtrate, respectively adding a saturated solution of sodium bicarbonate, washing with 1N diluted hydrochloric acid and saturated saline, separating an organic layer, drying, filtering, concentrating in vacuum, washing with a solvent to obtain kiwi fruit seed oil ceramide, and analyzing the product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linolenic acid-phytosphingosine ceramide 8.3min, linoleic acid-phytosphingosine ceramide 9.5min, palmitic acid-phytosphingosine ceramide 10.7min, oleic acid-phytosphingosine ceramide 11.3min, stearic acid-phytosphingosine ceramide 13.9min.

The obtained product is analyzed by high performance liquid chromatography, and the contents of linolenic acid-phytosphingosine ceramide, linoleic acid-phytosphingosine ceramide, oleic acid-phytosphingosine ceramide, palmitic acid-phytosphingosine ceramide and stearic acid-phytosphingosine ceramide are 56%, 20%, 12%, 5%, 2.5% and the balance of other components are less.

Example 14

Synthesis of ceramide from Malura oil fatty acid and sphinganine

The first step: 50g of Malawa oil was dissolved in 100mL of tetrahydrofuran, cooled in an ice bath, 150mL of potassium hydroxide (25 wt%) was added dropwise, and after the addition was completed, the reaction was allowed to stand at room temperature until TLC detection was completed.

Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 150mL of ethyl acetate to extract a water phase, adding 150mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase ₂ SO ₄ Drying, filtration and concentration in vacuo gave 43.1g of a marlura oil fatty acid.

And a second step of: malawa oil fatty acid (50 mmol, based on the main component fatty acid), DCC (80 mmol), DMAP (20 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, and then sphinganine (60 mmol) was added to the reaction system, followed by stirring at room temperature until TLC detection was complete.

Post-treatment: the reaction solution is filtered to remove solids, the filtrate is collected, the saturated solution of sodium bicarbonate, 1N diluted hydrochloric acid and saturated saline water are respectively added for washing, an organic layer is separated, dried, filtered and concentrated in vacuum, and then the product is washed by a solvent to obtain the Mala oil ceramide, and the product is analyzed by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c 3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The retention time of each component HPLC was: linoleic acid-dihydrosphingosine ceramide 9.4min, palmitic acid-dihydrosphingosine ceramide 10.6min, oleic acid-dihydrosphingosine ceramide 11.1min, stearic acid-dihydrosphingosine ceramide 13.5min, arachidic acid-dihydrosphingosine ceramide 16.0min.

The obtained products are analyzed by high performance liquid chromatography, the content of oleic acid-dihydrosphingosine ceramide, palmitic acid-dihydrosphingosine ceramide, linoleic acid-dihydrosphingosine ceramide, stearic acid-dihydrosphingosine ceramide and arachidic acid-dihydrosphingosine ceramide is 71%, 10%, 8%, 6.5% and 2% in sequence, and the balance is other components, and the content is less.

Example 15

MTT method for detecting proliferation activity of compound on cell

HaCaT cells were grown at 1X 10 ⁴ The density of individuals/wells was seeded in 96-well plates and the incubator was overnight. After 24h, the supernatant was discarded, 100. Mu.L of medium containing samples of different concentrations (product of example 1) was added, incubation was continued for 24h, medium was removed, 100. Mu.L of thiazole blue (MTT) was added to each well, absorbance at 450nm was measured, and cell viability = A was calculated _{Drug delivery hole} /A _{Blank hole} ×100％。

As shown in FIG. 1, sesame oil ceramide has a promoting effect on cell viability, and cell viability rates at concentrations of 3.90625, 7.8125, 15.625, 31.25, 62.5, 125, 250, 500, and 1000mg/L are 125.91%, 109.19%, 121.45%, 117.69%, 112.53%, 111.56%, 99.58%, 92.34%, and 71.45%, respectively. The effective concentration is as low as 4mg/L, the safe concentration is 250mg/L, the concentration gradient is stable, the obvious effect of promoting cell proliferation is shown, and the tissue repair capability is good.

Example 16

Assessment of skin barrier repair by cell migration

Principle of: when the cells grow to be fused into a single-layer state, a scratch tool is manufactured on the fused single-layer cells, the cells in the blank area are removed by mechanical force, the migration condition of the cells to the cell-free area is observed through a period of culture, and the migration capability of the cells is reflected by measuring the migration distance of the cells.

The operation steps are as follows:

1. the culture plate is streaked. Firstly, a Marker pen is used for uniformly scribing transverse lines by comparing with a straight ruler, and the transverse lines are crossed through the through holes at intervals of about 0.5 cm to 1cm, and each hole at least passes through 5 lines, so that attention lines are not too thick when scribing.

2. And (5) paving cells. About 5X 10 is added to the well ⁵ Individual cells (the number of different cells is different, and the cell growth speed is regulated), and the inoculation principle is that the fusion rate reaches 100% after overnight.

3. Cell streaking. The next day the tip is used to scratch the cell layer along the line marked on the back of the plate on the first day, perpendicular to the cell plane (the same tip is preferably used between the different wells).

4. Washing cells. After the streaking was completed, cells were washed 3 times with sterile PBS, cells that did not adhere to the wall, i.e., streaked cells at streaking, and the gap left after streaking was clearly visible, followed by replacement of fresh serum-free medium.

5. And (5) culturing and observing the cells. After the samples (products of examples 1 to 3 and ceramide 3B) were diluted with the medium (product concentration of example 1 was 20mg/L, product concentration of example 2, product concentration of example 3 and ceramide 3B concentration were all 100 mg/L), the cells were placed in a cell culture dish, and the cells were placed in 5wt% CO at 37 ℃C ₂ Incubator culture, after 24 hours, cells were removed, observed with a microscope and the width of scratches was measured, and photographed, and the healing rate was calculated using Image J software.

The results are shown in fig. 2, and the score width of the experimental group is narrower for sesame oil ceramide than for the solvent control group, indicating that sesame oil ceramide has better tissue healing ability. The solvent control group had a healing rate of 21.64% after 24 hours, sesame oil ceramide had a healing rate of 65.31% after 24 hours, and ceramide 3B had a healing rate of 59.32% after 24 hours. The compound obviously improves the cell healing rate, has good skin tissue repair activity and has better effect than ceramide 3B.

As shown in fig. 3, the scratch width was narrower for the corn oil ceramide than for the solvent control group, indicating that corn oil ceramide has better tissue healing ability. The solvent control group had a 32.18% healing rate after 24 hours, the corn oil ceramide had a 92.15% healing rate after 24 hours, and ceramide 3B had a 59.32% healing rate after 24 hours. The compound obviously improves the cell healing rate, has good skin tissue repair activity and has better effect than ceramide 3B.

As shown in fig. 4, the scratch width was narrower for the almond oil ceramide than for the solvent control group, indicating that the almond oil ceramide has better tissue healing ability. The solvent control group had a healing rate of 28.25 after 24 hours, almond oil ceramide had a healing rate of 63.47% after 24 hours, and ceramide 3B had a healing rate of 59.32% after 24 hours. The compound of the invention improves the cell healing rate, has good skin tissue repair activity and better effect than ceramide 3B.

Example 17

Elastase inhibition experiment tests anti-aging effect

Elastase inhibition method: 2mg/mL elastase solution (2 mL) is taken, samples with different concentrations (products of examples 4-6) are added, vortex mixing is fully carried out, shaking is carried out for 20min at 37 ℃ by a 400r/min shaking table, 5mL of 0.5mol/L phosphate buffer solution with pH of 6.0 is immediately added, vortex mixing is carried out, a proper amount of mixed solution is taken into a 2mL centrifuge tube, centrifugation is carried out for 10min at 9 391×g, 200 mu L of supernatant is precisely sucked into a 96-well plate, absorbance is measured by an enzyme-labeled instrument at a wavelength of 495nm, and spectrum scanning at 400-800 nm is carried out.

The substrate enzyme adding solution is used as a blank control group, the substrate enzyme adding and sample solution is used as an enzyme inhibition group, and the substrate enzyme adding and sample solution is used as a background. Each group is provided with 3 multiple holes. Inhibition ratio (%) = [1- (An-An ')/(A0-A0') ] ×100%, where A0 is absorbance with no enzyme added to the sample, A0 'is absorbance with no enzyme added to the substrate and no sample added to the enzyme, an is absorbance with only sample solution, an' is absorbance with no enzyme added to the sample. When An ' > An, the effect is expressed as acceleration, and the acceleration rate (%) = [1- (An ' -An)/(A0-A0 ') ] ×100%.

As shown in FIG. 5, the results show that the allium sativum oil ceramide has a good inhibitory effect on elastase at various concentrations, specifically, the inhibition rate of elastase at a concentration of 0.25g/L was 7.47%, the inhibition rate of elastase at a concentration of 0.5g/L was 16.33%, the inhibition rate of elastase at a concentration of 1.0g/L was 23.60%, and the inhibition rate of elastase at a concentration of 2.0g/L was 16.67%.

As shown in FIG. 6, peanut oil ceramide has good inhibition effect on elastase at different concentrations, specifically, the inhibition rate of elastase at the concentration of 0.25g/L is 10.47%, the inhibition rate of elastase at the concentration of 0.5g/L is 18.93%, the inhibition rate of elastase at the concentration of 1.0g/L is 28.33%, and the inhibition rate of elastase at the concentration of 2.0g/L is 40.67%.

As shown in FIG. 7, the blackcurrant oil ceramide has good inhibition effect on elastase at different concentrations, specifically, the inhibition rate of elastase at the concentration of 0.25g/L is 10.13%, the inhibition rate of elastase at the concentration of 0.5g/L is 19.27%, the inhibition rate of elastase at the concentration of 1.0g/L is 30.67%, and the inhibition rate of elastase at the concentration of 2.0g/L is 45.00%.

The inhibitory activity of ceramide 2 against elastase was measured in the same manner, and as a result, as shown in FIG. 8, the elastase inhibition rates at concentrations of 0.25, 0.5, 1.0 and 2.0g/L were 10.12%, 18.06%, 28.84% and 19.78%, respectively, which were not as effective as peanut oil ceramide and blackcurrant oil ceramide at the same concentrations.

Example 18

LPS induced cell method for detecting anti-inflammatory repair efficacy

B16 mouse melanoma cells were grown at a density of 1X 10 ⁴ The cells/wells were seeded in 96-well plates, placed in an incubator overnight, the supernatant was discarded after 24 hours, 100. Mu.L of samples of different concentrations diluted with DMEM medium (product of example 7) were added, the negative control group was DMEM medium without samples, 3 wells per group, and mixed with CO at 5 wt.% ₂ Incubate at 37 ℃. Lipopolysaccharide model group and experimental group were added with 10 μg/mL LPS and incubated together for 24h 2h after dosing. After the reaction, 50. Mu.L of the cell supernatant was collected, and the intracellular IL-6 gene expression was detected using an IL-6ELISA kit.

The results are shown in FIG. 9, where IL-6 levels were 10.95 times the basal levels at a working concentration of 10. Mu.g/mL of LPS stimulation. Under the action of the rapeseed oil ceramide with the concentration of 50mg/L, 100mg/L, 200mg/L and 400mg/L respectively, the IL-6 factor level is obviously reduced and is 0.87, 0.64, 0.52 and 0.16 times of that of an LPS model group respectively, and the rapeseed oil ceramide is dose-dependent, so that the rapeseed oil ceramide has good anti-inflammatory effect and can promote the repair of inflammatory damaged skin.

Example 19

MMP1 is also called interstitial collagenase and matrix metalloproteinase, belongs to matrix metalloproteinase family, and its main acting substrate is fibrous collagen, which can degrade collagen fiber and gelatin in extracellular matrix and change microenvironment of cells. MMP1 plays an important role in elastin, inhibiting MMP1 can improve the synthesis of fibroblast collagen and elastin, and reducing MMP activity can increase the collagen synthesis speed.

HaCaT cells were grown at 1X 10 ⁵ The density of individuals/wells was seeded in 96-well plates and the incubator was overnight. After 24h, the supernatant was discarded, 100. Mu.L of medium containing samples of different concentrations (products of examples 8 to 10) was added, modelThe group is not added with sample, the negative control group is DMEM culture medium without sample, 3 compound holes are formed in each group, and the mass fraction of CO is 5% ₂ After incubation for 2h at 37℃either UVA or UVB ultraviolet radiation is irradiated. The distance between the ultraviolet radiation source and the cells was 15cm, and the UVA intensity was 200mJ/cm ² The irradiation time was 2 hours, and the UVB intensity was 50mJ/cm ² The irradiation time was 1h. After the end of irradiation, incubation was continued for 12h in the incubator. Intracellular MMP-1 gene expression was detected using an MMP-1ELISA kit. Inhibition = 1- (experimental group MMP1 expression level/model group MMP1 expression level) ×100%.

The results are shown in FIGS. 10 and 11, european Li Ziyou ceramide: for UVA, the MMP1 expression level of the negative control group is set to be 1, the expression level of the model group is 1.90, and the inhibition rate of the MMP1 expression of the model group is 38%, 51% and 67% when the European Li Ziyou ceramide is at the concentration of 125, 250 and 400 mg/L; the expression level of MMP1 in the negative control group was 1, the expression level in the model group was 2.33, and the inhibition rate of MMP1 expression in the European Li Ziyou ceramide at 125, 250 and 400mg/L was 43%, 48% and 68% relative to the model group.

The results are shown in fig. 12 and 13, which show that hazelnut oil ceramide: for UVA, the MMP1 expression level of the negative control group is set to be 1, the expression level of the model group is 1.90, and the inhibition rate of the MMP1 expression of the hazelnut oil ceramide is 29%, 44% and 62% relative to the model group at the concentration of 125, 250 and 400 mg/L; for UVB, the MMP1 expression level of the negative control group was 1, the expression level of the model group was 2.33, and the inhibition rate of the MMP1 expression of the hazelnut oil ceramide was 35%, 50% and 68% relative to the model group at the concentrations of 125, 250 and 400 mg/L.

The results are shown in fig. 14 and 15, pumpkin seed oil ceramide: for UVA, the MMP1 expression level of the negative control group is set to be 1, the expression level of the model group is set to be 1.90, and the MMP1 expression level is 12%, 25% and 26% when the pumpkin seed oil ceramide is at the concentration of 125, 250 and 400 mg/L; for UVB, the MMP1 expression level of the negative control group was 1, the expression level of the model group was 2.33, and the MMP1 expression levels were 34%, 44% and 56% at concentrations of 125, 250 and 400mg/L of pumpkin seed oil ceramide.

After UVA uv radiation, keratinocytes promote elevated expression of MMP1 by fibroblasts, thereby causing degradation of the extracellular matrix of the skin and collagen of the skin, leading to photoaging of the skin. The results show that the vegetable oil ceramide can inhibit the fibroblast from producing MMP1 caused by ultraviolet radiation, and has a certain effect on preventing skin photoaging.

Example 20

DPPH free radical scavenging detection of antioxidant performance

DPPH is 1, 1-diphenyl-2-trinitrophenylhydrazine, and can be used for antioxidant experiments.

Samples (products of examples 10 to 12) with corresponding concentrations (50, 100, 200, 400, 800 mg/L) were mixed with 0.1mol/LDPPH, absolute ethanol solution according to a ratio of 1:1, and DPPH and absolute ethyl alcohol 1:1, and the absorbance at 517 nm. The absorbance of the sample and the reaction solution of DPPH was designated as A1, the absorbance of the sample and the reaction solution of absolute ethyl alcohol was designated as A2, the absorbance of the reaction solution of DPPH and absolute ethyl alcohol was designated as A3, and the clearance rate of DPPH of the sample was = [1- (A1-A2)/A3 ]. Times.100%.

The results are shown in fig. 16, pumpkin seed oil ceramide: DPPH radical scavenging rates at concentrations of 50, 100, 200, 400 and 800mg/L were 15.76%, 23.91%, 34.09%, 36.77% and 46.48%, respectively, and excellent antioxidant effects were exhibited. DHA algae oil ceramide (FIG. 17): DPPH radical scavenging rates at concentrations of 50, 100, 200, 400 and 800mg/L were 17.76%, 24.91%, 36.20%, 41.93% and 46.14%, respectively, and excellent antioxidation effects were exhibited. Calophyllum inophyllum oil ceramide (figure 18): DPPH radical scavenging rates at concentrations of 50, 100, 200, 400 and 800mg/L were 9.43%, 18.91%, 25.77%, 34.04% and 41.86%, respectively, and excellent antioxidant effects were exhibited.

The antioxidant effect of ceramide 3B (i.e., oleic acid ceramide) was measured in the same manner, and the result was shown in FIG. 19, in which DPPH radical scavenging rates at concentrations of 50, 100, 200, 400, 800mg/L were 7.76%, 12.82%, 24.10%, 29.60%, 33.16%. The clearance rate of the pumpkin seed oil ceramide, DHA algae oil ceramide and crabapple oil ceramide on DPPH is higher than that of ceramide 3B, and the pumpkin seed oil ceramide and the DHA algae oil ceramide have better antioxidation effect.

Example 21

Whitening Activity test

Taking B16 cells in exponential growth phase, digesting with trypsin-EDTA with mass fraction of 0.25%, blowing uniformly, and mixing the cells according to 3×10 ⁵ Density of individual/well was seeded in 12-well plates. At 37 ℃, the mass fraction of CO is 5 percent ₂ Incubated overnight in the environment. Removing supernatant, adding culture solution containing samples with different mass concentrations (products of examples 12-14), incubating with RPMI-1640 culture medium without sample as blank group, incubating with DMEM culture medium as mould group, and incubating with 3 compound holes each with mass fraction of 5% CO ₂ Incubation was carried out for 24h at 37 ℃. The culture medium in the well plate is discarded, and after washing with Phosphate Buffer (PBS) for one to two times, 1mL of NaOH solution (1 mol/L) containing 10% DMSO by mass fraction is added to lyse the cells, and the cells are kept at a constant temperature of 80℃or 100℃for 2 hours until the cells are completely lysed. The absorbance was measured at 405nm in a microplate reader. The melanin inhibition rate=1- (OD value per well/OD value of model group) ×100% was calculated.

As shown in fig. 20, the melanin content of the blank group was 1, the melanin expression of the model group was 1.51, and the melanin inhibition rates of the begonia fruit ceramide were 7.69%, 12.96%, 13.36%, 16.73% and 22.84% at concentrations of 10, 20, 40, 80 and 100mg/L, respectively, and a certain whitening effect was exhibited. As shown in fig. 21, the blank group had a melanin content of 1, and the manufactured group had a melanin expression of 1.51, and the kiwi fruit oil ceramide had melanin inhibition rates of 8.14%, 12.52%, 13.36%, 12.31%, and 20.63% at concentrations of 10, 20, 40, 80, and 100mg/L, respectively, and exhibited a certain whitening effect. As shown in fig. 22, the melanin content of the blank group was 1, the melanin expression of the model group was 1.54, and the melanin inhibition rates of the malva oil ceramide were 15.53%, 22.42%, 30.82%, 40.39%, 47.02% at concentrations of 10, 20, 40, 80, and 100mg/L, respectively, and excellent whitening effect was exhibited.

The foregoing is merely illustrative embodiments of the present invention, and the present invention is not limited thereto, and any changes or substitutions that may be easily contemplated by those skilled in the art within the scope of the present invention should be included in the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope of the claims.

Claims

1. Vegetable oil ceramide, which is obtained by reacting vegetable oil fatty acid with sphingosine compound, wherein the sphingosine compound is selected from sphingosine, phytosphingosine and dihydrosphingosine, and the vegetable oil fatty acid is selected from sesame oil fatty acid, corn oil fatty acid, almond oil fatty acid, garlic oil fatty acid, peanut oil fatty acid, blackcurrant seed oil fatty acid, rapeseed oil fatty acid, hazelnut oil fatty acid, pumpkin seed oil fatty acid, DHA algae oil fatty acid, crab apple oil fatty acid, kiwi seed oil fatty acid, malva oil fatty acid and European Li Ziyou fatty acid.

2. The vegetable oil ceramide of claim 1, wherein the vegetable oil fatty acid is obtained by hydrolysis of vegetable oil.

3. The vegetable oil ceramide of claim 1, wherein the sesame oil fatty acid has a composition of: 45-70 wt% of linoleic acid, 15-35 wt% of oleic acid, 6-20 wt% of palmitic acid, 0-2 wt% of stearic acid, 0-1 wt% of arachidic acid and 0-0.5 wt% of linolenic acid; the composition of the corn oil fatty acid is as follows: 50 to 70 weight percent of linoleic acid, 20 to 35 weight percent of oleic acid, 5 to 18 weight percent of palmitic acid, 0.5 to 3 weight percent of stearic acid, 0.01 to 1 weight percent of linolenic acid and 0 to 1 weight percent of arachidic acid; the almond oil fatty acid comprises the following components: 55 to 80 weight percent of oleic acid, 15 to 35 weight percent of linoleic acid, 0.1 to 8 weight percent of palmitic acid, 0.5 to 2 weight percent of stearic acid and 0.01 to 1 weight percent of linolenic acid; the composition of the garlic oil fatty acid is as follows: 40-60 wt% of nervonic acid, 30-50 wt% of oleic acid, 5-12 wt% of arachidic acid, 0-2 wt% of palmitic acid and 0-2 wt% of linoleic acid; the peanut oil fatty acid comprises the following components: 40 to 60 weight percent of linoleic acid, 30 to 50 weight percent of oleic acid, 4 to 12 weight percent of palmitic acid, 0.5 to 5 weight percent of stearic acid, 0 to 1 weight percent of arachidic acid and 0 to 1 weight percent of behenic acid; the blackcurrant seed oil fatty acid comprises the following components: 40-60 wt% of linoleic acid, 20-40 wt% of linolenic acid, 10-20 wt% of oleic acid, 1-8 wt% of palmitic acid, 0-2 wt% of stearic acid and 0-2 wt% of arachidic acid; the composition of the rapeseed oil fatty acid is as follows: 50 to 80 weight percent of oleic acid, 10 to 30 weight percent of linoleic acid, 2 to 8 weight percent of linolenic acid, 1 to 5 weight percent of stearic acid, 0.5 to 5 weight percent of palmitic acid, 0 to 1 weight percent of arachidic acid, 0 to 1 weight percent of arachidonic acid and 0 to 1 weight percent of erucic acid; the hazelnut oil fatty acid comprises the following components: 70 to 90 weight percent of oleic acid, 8 to 25 weight percent of linoleic acid, 0.5 to 5 weight percent of palmitic acid, 0.5 to 4 weight percent of stearic acid, 0 to 0.5 weight percent of linolenic acid and 0 to 0.5 weight percent of palmitoleic acid; the pumpkin seed oil fatty acid comprises the following components: 40 to 70 weight percent of linoleic acid, 20 to 55 weight percent of oleic acid, 3 to 10 weight percent of palmitic acid, 1 to 5 weight percent of stearic acid, 0.1 to 5 weight percent of linolenic acid, 0 to 1 weight percent of arachic acid and 0 to 0.5 weight percent of palmitoleic acid; the DHA algae oil fatty acid comprises the following components: 50 to 75 weight percent of DHA, 15 to 35 weight percent of oleic acid, 3 to 15 weight percent of palmitic acid, 0.1 to 1 weight percent of stearic acid, 0.1 to 1 weight percent of EPA, 0.01 to 0.5 weight percent of linolenic acid, 0 to 0.5 weight percent of arachidic acid and 0 to 0.5 weight percent of behenic acid; the composition of the crab apple oil fatty acid is as follows: 30 to 50 weight percent of oleic acid, 15 to 40 weight percent of linoleic acid, 8 to 20 weight percent of stearic acid, 6 to 20 weight percent of palmitic acid, 0.2 to 1.5 weight percent of arachidic acid, 0.05 to 2 weight percent of linolenic acid, 0.05 to 1 weight percent of palmitoleic acid, 0 to 1 weight percent of behenic acid and 0 to 0.5 weight percent of arachidonic acid; the kiwi seed oil fatty acid comprises the following components: 45-70 wt% of linolenic acid, 8-20 wt% of linoleic acid, 5-20 wt% of oleic acid, 2-10 wt% of palmitic acid and 0.5-5 wt% of stearic acid; the composition of the Malara oil fatty acid is 65-85 wt% of oleic acid, 5-15 wt% of palmitic acid, 2-10 wt% of linoleic acid, 2-10 wt% of stearic acid, 0.2-2 wt% of arachidic acid, 0-1 wt% of arachidonic acid and 0-0.5 wt% of linolenic acid; the composition of the European Li Ziyou fatty acid is as follows: 60 to 80 weight percent of oleic acid, 10 to 30 weight percent of linoleic acid, 0.5 to 5 weight percent of stearic acid, 0.5 to 3 weight percent of palmitoleic acid, 0 to 7 weight percent of palmitic acid and 0 to 1 weight percent of linolenic acid.

4. Vegetable oil ceramide, its composition includes: oleic acid ceramide, linoleic acid ceramide, palmitic acid ceramide.

5. The vegetable oil ceramide of claim 4, which comprises the following components: 10 to 90 weight percent of oleic acid ceramide, 2 to 70 weight percent of linoleic acid ceramide and 0.1 to 20 weight percent of palmitic acid ceramide.

6. The vegetable oil ceramide of claim 4 or 5, further comprising at least one of linolenic acid ceramide, DHA ceramide, nervonic acid ceramide, arachidic acid ceramide, stearic acid ceramide, behenic acid ceramide, arachidonic acid ceramide, erucic acid ceramide, palmitoleic acid ceramide, EPA ceramide.

7. The synthetic method of the vegetable oil ceramide of any one of claims 1 to 6, comprising the following steps:

under the conditions of condensing agent and organic alkali, vegetable oil fatty acid reacts with sphingosine compound, the condensing agent is DCC, and the organic alkali is DMAP;

the molar ratio of the vegetable oil fatty acid to the sphingosine compound to the DCC to the DMAP is 1: (1-1.5): (1-2): (0.2-0.5), wherein the solvent for the reaction is at least one of dichloromethane, tetrahydrofuran, ethyl acetate and acetonitrile.

8. Use of the vegetable oil ceramide of any one of claims 1 to 6 in cosmetics, pharmaceuticals, dietary or health products.

9. The use according to claim 8, wherein said vegetable oil ceramide has at least one of skin barrier repair, tissue healing, anti-aging, anti-inflammatory, anti-photoaging, antioxidant, promoting collagen synthesis, maintaining elastin viability, whitening efficacy.

10. A composition comprising the vegetable oil ceramide of any one of claims 1-6, which has at least one of skin barrier repair, tissue healing, anti-aging, anti-inflammatory, anti-photoaging, antioxidant, promotion of collagen synthesis, maintenance of elastin activity, whitening efficacy.