CN115974714A

CN115974714A - Shea butter ceramide, and its synthesis method and use

Info

Publication number: CN115974714A
Application number: CN202310051553.9A
Authority: CN
Inventors: 杨超文; 叶柳
Original assignee: Shenzhen Dikeman Biotechnology Co ltd
Current assignee: Shenzhen Dikeman Biotechnology Co ltd
Priority date: 2023-02-02
Filing date: 2023-02-02
Publication date: 2023-04-18

Abstract

The invention belongs to the technical field of biological medicines, and discloses shea butter ceramide, which is obtained by reacting shea butter fatty acid with sphingoid compounds selected from sphingosine, phytosphingosine and dihydrosphingosine. Shea butter ceramide has excellent performances in the aspects of repairing natural barrier of skin, healing tissue, resisting aging and the like, and has wide application prospect in the fields of cosmetics, health care products, biological medicines and the like.

Description

Shea butter ceramide, and its synthesis method and use

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to shea butter ceramide, a synthesis method and application thereof.

Background

Ceramide (also called molecular nail) naturally exists in skin, is a very important component of skin barrier (stratum corneum) and is up to 40-50 wt%, and is a sphingolipid consisting of sphingoid long-chain bases and fatty acids, wherein the carbon chain length, unsaturation degree and hydroxyl number of the sphingosine part and the fatty acid part can be changed, and the Ceramide represents a compound. Ceramides exhibit excellent properties in regulating skin barrier function, restoring skin moisture, and enhancing adhesion between skin keratinocytes, etc.

Due to 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 characteristics of more sustainable and more environment-friendly raw material sources and the similar components of the ceramide and the ceramide of the skin, and can become a next-generation environment-friendly, safe and reliable ceramide product.

Shea butter is extracted from the fruit of Shea butter, which is mainly grown in the dilute grassland of West Africa, zhongfei and Dongfei. Fresh shea butter is yellow paste with a characteristic odor, and usually requires refining to obtain a raw material for cosmetics. Like most vegetable oils, shea butter also contains triglycerides as the major component, and the fatty acid moieties are mainly saturated fatty acids such as palmitic acid, stearic acid, arachidic acid, and the like, and unsaturated fatty acids such as oleic acid, linoleic acid, and the like. In addition, the shea butter also contains 7-10 wt% of unsaponifiable matter, and the unsaponifiable matter mainly comprises phytosterol and tocopherol (vitamin E). In addition, triterpene esters are also important components of shea butter. The shea butter has excellent skin care effect due to a large amount of unsaturated fatty acid and unsaponifiable components. Shea butter has excellent effects in moisturizing, resisting aging, diminishing inflammation, and strengthening skin barrier function.

Disclosure of Invention

The invention aims to provide ceramide synthesized by using shea butter fatty acid of plant origin.

Another object of the present invention is to provide a method for synthesizing shea butter ceramide, which uses shea butter oil or shea butter fatty acid, which is easily available and is derived from natural plants, as a raw material.

Another object of the present invention is to provide the use of shea butter ceramide.

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

in a first aspect of the invention, shea butter ceramides, obtained by reacting shea butter fatty acids with sphingoid compounds selected from sphingosine, phytosphingosine, dihydrosphingosine.

The reaction can be a chemical synthesis reaction (as detailed below), or a microbial fermentation method, namely, pichia pastoris or saccharomyces cerevisiae is used for fermentation under certain environment to obtain sphingoid compounds, and then fatty acid is added to finally obtain ceramide; or taking shea butter as a raw material, selecting a proper strain, and fermenting to obtain shea butter ceramide.

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

Further, the shea butter fatty acid is obtained by hydrolyzing shea butter oil.

Further, the shea butter fatty acid contains 35 to 60wt% of oleic acid.

Further, the shea butter fatty acid contains 25-55 wt% of stearic acid.

Further, the shea butter fatty acid contains 2-10 wt% of linoleic acid.

Further, the shea butter fatty acid contains 0.5 to 10wt% of palmitic acid.

Further, the shea butter fatty acid contains 0.1-2 wt% of arachidic acid.

In addition, the shea butter fatty acid also contains 0-1 wt% of linolenic acid and 0-1 wt% of behenic acid.

The shea butter fatty acid comprises the following components: 35 to 60 weight percent of oleic acid, 25 to 55 weight percent of stearic acid, 2 to 10 weight percent of linoleic acid, 0.5 to 10 weight percent of palmitic acid, 0.1 to 2 weight percent of arachidic acid, 0 to 1 weight percent of linolenic acid and 0 to 1 weight percent of behenic acid.

The main components of the shea butter fatty acid are oleic acid and stearic acid, other fatty acids comprise linoleic acid, palmitic acid and arachidic acid, which are essential components, the content of each component can be different under the influence of plant varieties, soil, climate, production area, picking season and extraction process, and the linolenic acid and the behenic acid are optional components or unnecessary components.

Shea butter ceramides, the composition of which comprises: oleic acid ceramide, stearic acid ceramide, linoleic acid ceramide, palmitic acid ceramide, arachidic acid ceramide; because fatty acids all participate in the same reaction, the mass ratio of ceramide after the reaction is not changed greatly, and therefore, similar to the composition of shea butter fatty acid, the shea butter ceramide composition is: 35 to 60 weight percent of oleic acid ceramide, 25 to 55 weight percent of stearic acid ceramide, 2 to 10 weight percent of linoleic acid ceramide, 0.5 to 10 weight percent of palmitic acid ceramide and 0.1 to 2 weight percent of arachidic acid ceramide. The content of each component is different due to different contents of fatty acid in shea butter or oil. In addition, the shea butter ceramide also comprises ceramide obtained by reacting one or more of linolenic acid and behenic acid with sphingoid compounds, namely, 0-1 wt% of linolenic acid ceramide and 0-1 wt% of behenic acid ceramide. Shea butter ceramides also include compounds such as phytosterols, tocopherols, triterpene esters, etc. that are present in Shea butter fatty acids but do not react with sphingoid compounds.

Shea butter ceramides, the composition of which comprises: oleic acid ceramide, stearic acid ceramide, linoleic acid ceramide; 35-60 wt% of oleic acid ceramide, 25-55 wt% of stearic acid ceramide and 2-10 wt% of linoleic acid ceramide.

Furthermore, the shea butter ceramide comprises 0.5-10 wt% of palmitic acid ceramide.

Furthermore, the shea butter ceramide comprises arachidic ceramide, and the content of the arachidic ceramide is 0.1-2 wt%.

Further, the shea butter ceramide comprises not more than 1wt% of linolenic acid ceramide and not more than 1wt% of behenic acid ceramide, especially 0.1 to 1wt% of linolenic acid ceramide and 0.1 to 1wt% of behenic acid ceramide.

The oleic acid ceramide is obtained by condensation reaction of oleic acid and sphingoid compounds, and comprises oleic acid phytosphingosine ceramide, oleic acid sphingosine ceramide and oleic acid dihydrosphingosine ceramide; the stearic acid ceramide is obtained by condensation reaction of stearic acid and sphingoid compounds, and comprises stearic acid phytosphingosine ceramide, stearic acid sphingosine ceramide and stearic acid dihydrosphingosine ceramide; linoleic acid ceramide is obtained by condensation reaction of linoleic acid and sphingoid compounds, and comprises linoleic acid phytosphingosine ceramide, linoleic acid sphingosine ceramide, and linoleic acid dihydrosphingosine ceramide; palmitic acid ceramide, arachidic acid ceramide, linolenic acid ceramide, behenic acid ceramide, and the like.

In a second aspect of the present invention, a method for synthesizing shea butter ceramide comprises the following steps:

under the conditions of a condensing agent and organic base, shea butter fatty acid reacts with a sphingoid compound, wherein the condensing agent is EDCI, and the organic base is DIPEA.

Further, the molar ratio of the shea butter fatty acid to the sphingoid compound to the EDCI to the DIPEA is 1: (1-1.5): (1-2): (1-2), wherein the solvent for reaction is at least one of dichloromethane, tetrahydrofuran, ethyl acetate and acetonitrile.

The shea butter purchased in the market is generally in the form of grease and needs to be hydrolyzed into shea butter fatty acid, so that the method further comprises the following steps:

the shea butter fatty acid is obtained by hydrolyzing shea butter oil through saponification reaction.

Further, the saponification reaction is hydrolysis of the shea butter oil in a potassium hydroxide solution.

Further, the mass ratio of the shea butter oil to the potassium hydroxide is 1: (1-2).

In a third aspect of the invention, the use of shea butter ceramide in cosmetics, pharmaceuticals, dietary foods or health products.

Further, the shea butter ceramide has at least one of skin barrier repair, tissue healing, anti-aging, anti-photoaging, anti-oxidation, collagen synthesis promotion, elastin vitality maintenance, and whitening effects.

A composition comprising shea butter ceramide, said composition having at least one of skin barrier repair, tissue healing, anti-aging, anti-photoaging, anti-oxidant, collagen synthesis promotion, elastin viability maintenance, and whitening efficacy.

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

The invention has the following beneficial effects:

shea butter fatty acid belongs to naturally formed fatty acid, the main components are oleic acid and stearic acid, in addition, linoleic acid, palmitic acid and arachidic acid are also contained, and the shea butter ceramide is prepared by mild reaction with sphingoid compounds naturally existing in skin, shows excellent performance in the aspects of repairing, antioxidation, anti-aging and the like of natural barriers of the skin, and has wide application prospect in the fields of cosmetics, health care products, biological medicines and the like.

1. Compared with single ceramide, the effect is better. Different ceramides have different effects due to their different structures, and ceramides with a single structure generally have poor comprehensive effects. The technical scheme is based on a bionic idea, shea butter oil or fatty acid from natural sources is used as a raw material to synthesize composite ceramide, so that the difference of the efficacies of different ceramides is made up, and trace amount of ceramide can be formed by trace amount of fatty acid in shea butter, so that the function supplement effect is achieved.

2. Compared with compounded ceramide, the effect is better. In addition to fatty acids (or oils), shea butter also contains various active ingredients such as phytosterols, tocopherols, triterpene esters, etc., and these nutrients have the effects of moistening skin, keeping moisture, etc. The ceramide synthesized by shea butter has synergistic effect with other active ingredients in shea butter, and has better effect compared with ceramide compounded according to similar proportion.

3. The cost is lower. The method of the invention can quickly obtain the composition compounded by various ceramides, the shea butter oil or fatty acid thereof of plant source has wide source, easy commercial acquisition, lower cost, more environmental protection and economy, and is different from the idea of mixing and compounding different single ceramides, the fatty acid of single component has high raw material price, and different ceramides are required to be respectively produced and then compounded, thereby increasing the preparation cost.

4. The synthetic 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 shows the results of the cell migration ability test in example 4;

FIG. 2 is a bar graph of the elastase inhibition of example 5;

FIGS. 3 and 4 are graphs showing MMP1 expression levels in the anti-photoaging test of example 6;

FIG. 5 is a bar graph of DPPH radical scavenging for oxidation resistance test of example 7;

fig. 6 is a bar graph of melanin content in the whitening activity test of example 8.

Detailed Description

The present invention will be further described with reference to the following specific examples.

EDCI refers to 1-ethyl- (3-dimethylaminopropyl) carbodiimide and DIPEA refers to N, N-diisopropylethylamine. The silica gel column chromatography uses Qingdao marine silica gel (particle size 0.040-0.063 mm). Thin Layer Chromatography (TLC) was performed using 60F254 silica gel plates, and TLC developed using UV light (254 nm) or iodine.

Example 1

Synthesis of ceramide from shea butter fatty acid and phytosphingosine

The first step is as follows: dissolving 50g of shea butter fat in 80mL of tetrahydrofuran, cooling in ice bath, dropwise adding 100mL of potassium hydroxide (25 wt%) solution, heating to room temperature after dropwise adding, and reacting until the TLC detection reaction is finished.

And (3) post-treatment: adding dilute hydrochloric acid (3N) to adjust pH to 3, adding 120mL ethyl acetate to extract water phase, adding 100mL saturated salt water to wash once, adding anhydrous Na to organic phase ₂ SO ₄ Drying, filtration and concentration in vacuo gave 41g of shea butter fatty acid.

The second step is that: adding shea butter fatty acid (50 mmol, calculated as main component fatty acid), EDCI (70 mmol) and DIPEA (70 mmol) into a 250mL round-bottom flask, adding 100mL dichloromethane, stirring at room temperature for 1 hour, adding phytosphingosine (70 mmol) into the reaction system, and stirring at room temperature until TLC detection reaction is finished.

And (3) post-treatment: adding water to quench and react, separating an organic layer, drying, filtering, concentrating in vacuum, washing by a solvent to obtain shea butter ceramide, analyzing a product by HPLC, and carrying out HPLC chromatographic conditions: using Shimadzu high performance liquid chromatograph (LC-2030C3D Plus), the column temperature was measured using Innoval ODS-2.6 × 250mm,5 μm column: 30 ℃, injection volume: 10 μ L, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The HPLC retention time of each component was: linolenic acid-phytosphingosine ceramide 8.6min, linoleic acid-phytosphingosine ceramide 9.5min, palmitic acid-phytosphingosine ceramide 10.7min, oleic acid-phytosphingosine ceramide 11.3min, stearic acid-phytosphingosine ceramide 13.8min, arachidic acid-phytosphingosine ceramide 16.5min, and behenic acid-phytosphingosine ceramide 18.3min.

According to the analysis of the obtained product by high performance liquid chromatography, the contents of oleic acid-phytosphingosine ceramide, stearic acid-phytosphingosine ceramide, linoleic acid-phytosphingosine ceramide, palmitic acid-phytosphingosine ceramide, arachidic acid-phytosphingosine ceramide, linolenic acid-phytosphingosine ceramide and behenic acid-phytosphingosine ceramide are 57%, 25%, 8%, 4%, 1% and 0.5% in sequence, and the rest is other components with less content.

Example 2

Synthesis of ceramide from shea butter fatty acid and sphingosine

And (3) post-treatment: adding dilute hydrochloric acid (3N) to adjust pH to 3, adding 120mL ethyl acetate to extract water phase, adding 100mL saturated salt water to wash once, adding anhydrous Na to organic phase ₂ SO ₄ Drying, filtration and concentration in vacuo gave 40.7g shea butter fatty acid.

The second step is that: shea butter fatty acid (50 mmol, based on the main fatty acid), EDCI (65 mmol), DIPEA (65 mmol) were charged into a 250mL round-bottomed flask, 100mL of dichloromethane was added, followed by stirring at room temperature for 1 hour, and then sphingosine (60 mmol) was added to the reaction system, followed by stirring at room temperature, until TLC detection was complete.

And (3) post-treatment: adding water to quench and react, separating an organic layer, drying, filtering, concentrating in vacuum, washing by a solvent to obtain shea butter ceramide, analyzing a product by HPLC, and carrying out HPLC chromatographic conditions: using an Shimadzu high performance liquid chromatograph (LC-2030C3D Plus), the mixture was purified by using an Innoval ODS-2.6 × 250mm,5 μm column, column temperature: 30 ℃, injection volume: 10 μ L, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.

The HPLC retention time of each component 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, arachidic acid-sphingosine ceramide 17.8min.

The obtained product is analyzed by high performance liquid chromatography, and the contents of oleic acid-sphingosine ceramide, stearic acid-sphingosine ceramide, linoleic acid-sphingosine ceramide, palmitic acid-sphingosine ceramide and arachidic acid-sphingosine ceramide are respectively 37%, 48%, 3%, 7% and 2%, and the rest is other components with less content.

Example 3

Synthesis of ceramide from shea butter fatty acid and sphinganine

The first step is as follows: dissolving 50g of shea butter oil in 80mL of tetrahydrofuran, cooling in ice bath, dropwise adding 100mL of potassium hydroxide (25 wt%) solution, heating to room temperature after dropwise adding, and reacting until TLC detection reaction is finished.

And (3) post-treatment: adding dilute hydrochloric acid (3N) to adjust pH to 3, adding 120mL ethyl acetate to extract water phase, adding 100mL saturated salt water to wash once, adding anhydrous Na to organic phase ₂ SO ₄ Drying, filtration and concentration in vacuo gave 41.2g shea butter fatty acid.

The second step: shea butter fatty acid (50 mmol, based on the main fatty acid), EDCI (80 mmol), DIPEA (80 mmol) were charged into a 250mL round-bottomed flask, 100mL of methylene chloride was added, followed by stirring at room temperature for 1 hour, and then dihydrosphingosine (55 mmol) was added to the reaction system, followed by stirring at room temperature, until TLC detection was completed.

The HPLC retention time of each component was: linolenic acid-sphinganine ceramide 8.2min, linoleic acid-sphinganine ceramide 9.4min, palmitic acid-sphinganine ceramide 10.6min, oleic acid-sphinganine ceramide 11.2min, stearic acid-sphinganine ceramide 13.5min, arachidic acid-sphinganine ceramide 16.0min, and behenic acid-sphinganine ceramide 21.9min.

The obtained product is analyzed by high performance liquid chromatography, the contents of oleic acid-sphinganine ceramide, stearic acid-sphinganine ceramide, linoleic acid-sphinganine ceramide, palmitic acid-sphinganine ceramide, arachidic acid-sphinganine ceramide, linolenic acid-sphinganine ceramide and behenic acid-sphinganine ceramide are respectively 44%, 33%, 10%, 8%, 1%, 0.5% and 1%, and the rest is other components with less content.

Example 4

Evaluation of skin Barrier repair by cell migration

The principle is as follows: when the cells grow to be fused into a monolayer, a scratch tool is used for manufacturing a blank area on the fused monolayer, the cells in the blank area are removed by mechanical force, the migration of the cells to a cell-free area is observed through a period of culture, and the migration capacity of the cells is reflected by measuring the migration distance of the cells.

The method comprises the following operation steps:

1. the plates were streaked. Firstly, a Marker pen is used at the back of a 6-hole plate, a straight ruler is used for uniformly drawing transverse lines, the transverse lines are drawn approximately every 0.5-1 cm, the transverse lines cross through holes, at least 5 lines pass through each hole, and the attention lines are not too thick when the lines are drawn.

2. And (5) laying cells. About 5X 10 additions to the wells ⁵ And (3) inoculating each cell (the number of different cells is different and is adjusted according to the growth speed of the cells), wherein the inoculation principle is that the fusion rate reaches 100 percent after the overnight inoculation.

3. And (4) cell lineation. The next day, the cell layer was scored with a tip perpendicular to the cell plane along the line drawn on the back of the plate on the first day (preferably the same tip is used between different wells).

4. The cells were washed. After the scoring was completed, cells were washed 3 times with sterile PBS, non-adherent cells were washed away, i.e., cells streaked during streaking, leaving a clear gap after streaking, and then fresh serum-free medium was replaced.

5. And (5) culturing and observing cells. Samples (example 1 product, ceramide 3B) were diluted with medium (example 1 product concentration 100mg/L, ceramide 3B concentration 100 mg/L) and added to a cell culture dish, and the cells were placed at 37 ℃ and 5wt% CO ₂ The cells were cultured in an incubator, taken out after 24 hours, observed and measured for width of scratch by a microscope, photographed, and the healing rate was calculated with Image J software.

The results are shown in fig. 1, and the scratch width of the experimental group is narrower than that of the solvent control group, which indicates that shea butter ceramide has better tissue healing capability. The healing rate of the solvent control group after 24h was 21.57%, the healing rate of shea butter ceramide after 24h was 75.92%, and the healing rate of ceramide 3B after 24h was 59.32%. The compound provided by the invention obviously improves the cell healing rate, has good skin tissue repair activity, and has a better effect than ceramide 3B.

Example 5

Elastase inhibition experiment for testing anti-aging effect

Elastase inhibition methods: 2mL of 2mg/mL elastase solution is taken, samples (products in example 1) with different concentrations are added, the mixture is fully and evenly mixed in a vortex mode, the mixture is shaken in a shaking table with 400r/min at 37 ℃ for 20min, 5mL of 0.5mol/L phosphate buffer solution with pH6.0 is immediately added, the mixture is evenly mixed in a vortex mode, a proper amount of evenly mixed solution is taken to be put into a 2mL centrifugal tube, the centrifugal tube is centrifuged for 10min at 9 391 Xg, 200 mu L of supernatant is precisely absorbed into a 96-well plate, absorbance is measured by an enzyme-labeling instrument at the wave length of 495nm, and spectrum scanning of 400-800 nm is carried out at the same time.

And taking a substrate and enzyme solution as a blank control group, taking a substrate and enzyme solution and a sample solution as an enzyme inhibition group, and taking the substrate and sample without the enzyme solution as background. Each group is provided with 3 holes. Inhibition (%) = [1- (An-An ')/(A0-A0') ] × 100%, where A0 is the absorbance of a sample with no enzyme, A0 'is the absorbance of a sample with no substrate and no enzyme, an is the absorbance of a solution with only a sample, and An' is the absorbance of a sample with no enzyme. If An ' > An, a promoting effect is exhibited, and the promoting rate (%) = [1- (An ' -An)/(A0-A0 ') ] × 100%.

As shown in FIG. 2, shea butter ceramide has a certain inhibitory effect on elastase at various concentrations, specifically, it has an inhibitory rate of 7.47% at a concentration of 0.25g/L, 16.67% at a concentration of 0.5g/L, 13.27% at a concentration of 1.0g/L, and 6.67% at a concentration of 2.0 g/L.

Example 6

MMP1 is also called interstitial collagenase and matrix metalloproteinase, belongs to a family of matrix metalloproteinase, and has the main action substrates of fibrous collagen, can degrade collagen fibers and gelatin in extracellular matrix and change the microenvironment of cells. MMP1 plays an important role in elastin, inhibition of MMP1 can improve synthesis of collagen and elastin of fibroblasts, and reduction of MMP activity can increase collagen synthesis speed.

HaCaT cells were cultured at 1X 10 ⁵ The density of cells/well was plated in 96-well plates overnight in an incubator. After 24h, the supernatant was discarded, 100. Mu.L of medium containing samples (product of example 1) at different concentrations was added, no sample was added to the model group, the negative control group was DMEM medium without sample, 3 wells each, and CO was added at a mass fraction of 5% ₂ After incubation at 37 ℃ for 2h, UVA or UVB ultraviolet radiation was applied. The distance between the ultraviolet radiation light source and the cell was 15cm, and the UVA intensity was 200mJ/cm ² The radiation time is 2h, and the UVB intensity is 50mJ/cm ² The irradiation time is 1h. After the irradiation was finished, incubation was continued in the incubator for 12h. MMP-1 gene expression in cells was detected using an MMP-1ELISA kit. The inhibition rate =1- (MMP 1 expression amount in experimental group/MMP 1 expression amount in model group) × 100%.

As shown in fig. 3 and 4, the expression level of MMP1 in the negative control group was 1, the expression level in the model group was 1.90, and the inhibition rate of shea butter ceramide at concentrations of 125, 250, and 400mg/L to MMP1 expression in the model group was 34%, 44%, and 56%; in UVB, the MMP1 expression level of the negative control group is 1, the expression level of the model group is 2.33, and the inhibition rate of shea butter ceramide at the concentration of 125, 250 and 400mg/L relative to the MMP1 expression of the model group is 28%, 40% and 58%.

After UVA ultraviolet radiation, keratinocytes promote increased fibroblast MMP1 expression, thereby causing degradation of skin extracellular matrix and skin collagen, resulting in skin photoaging. The results show that the shea butter ceramide can inhibit the fibroblast caused by ultraviolet radiation from generating MMP1, and has certain effect on preventing skin photoaging.

Example 7

DPPH free radical scavenging and detecting antioxidant performance

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

Samples (product of example 1) at the corresponding concentrations (50, 100, 200, 400, 800 mg/L) were mixed with 0.1mol/L DPPH, absolute ethanol solution at a molar ratio of 1:1, mixing the mixture evenly, mixing DPPH and absolute ethyl alcohol 1:1, mixing the mixture in equal volume, reacting the mixture for 30min in a dark place at room temperature, and measuring the light absorption value at 517 nm. The absorbance of the sample and the DPPH reaction solution was denoted as A1, the absorbance of the sample and the absolute ethanol reaction solution was denoted as A2, the absorbance of the DPPH and the absolute ethanol reaction solution was denoted as A3, and the DPPH clearance of the sample = [1- (A1-A2)/A3 ] × 100%.

As shown in FIG. 5, the DPPH radical removal rates at concentrations of 50, 100, 200, 400, and 800mg/L were 5.56%, 9.44%, 12.93%, 17.00%, and 20.72%, respectively, and the antioxidant effect was excellent.

Example 8

Whitening Activity test

Taking B16 cells in exponential growth phase, digesting with trypsin-EDTA with the mass fraction of 0.25%, blowing uniformly, and mixing the cells according to the proportion of 3 multiplied by 10 ⁵ The density of each well was seeded in 12-well plates. At 37 ℃ and 5% by mass of CO ₂ Incubate overnight in the environment. Discarding supernatant, adding samples containing different mass concentrationsThe culture solution of the product (product of example 1) was incubated in RPMI-1640 medium without addition of the sample as a blank group and in DMEM medium as a molding group, and 3 duplicate wells were each incubated, and the content of CO was 5% by mass ₂ And incubating for 24h at 37 ℃. After discarding the medium from the well plate, washing once to twice with Phosphate Buffered Saline (PBS), 1mL of NaOH solution containing 10% by mass of DMSO (1 mol/L) was added to lyse the cells, and the cells were left to stand at 80 ℃ or 100 ℃ for 2 hours until they were completely lysed. The sample was placed in a microplate reader and absorbance was measured at 405 nm. Melanin inhibition =1- (each well OD value/model group OD value) × 100% was calculated.

As shown in fig. 6, the melanin content of the blank control group was 1, the melanin expression of the modeling group was 1.51, and the melanin inhibition ratios of shea butter ceramide were 8.14%, 14.73%, 22.21%, 27.79% and 38.33%, respectively, at concentrations of 10, 20, 40, 80 and 100mg/L, thereby exhibiting excellent whitening effects.

The above description is only an embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are 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. Shea butter ceramides, obtainable by reacting shea butter fatty acids with sphingoid compounds selected from sphingosine, phytosphingosine, dihydrosphingosine.

2. Shea butter ceramide according to claim 1, wherein the shea butter fatty acid is obtained by hydrolysis of shea butter.

3. Shea butter ceramide according to claim 1 or 2, characterized in that shea butter fatty acids contain 35-60 wt% oleic acid, 25-55 wt% stearic acid, 2-10 wt% linoleic acid, 0.5-10 wt% palmitic acid, 0.1-2 wt% arachidic acid.

4. Shea butter ceramides, the composition of which comprises: oleic acid ceramide, stearic acid ceramide, linoleic acid ceramide, palmitic acid ceramide, arachidic acid ceramide.

5. Shea butter ceramides according to claim 4, the composition of which comprises: 35 to 60 weight percent of oleic acid ceramide, 25 to 55 weight percent of stearic acid ceramide, 2 to 10 weight percent of linoleic acid ceramide, 0.5 to 10 weight percent of palmitic acid ceramide and 0.1 to 2 weight percent of arachidic acid ceramide.

6. Shea butter ceramide according to claim 4 or 5, whose composition further comprises: 0 to 1 weight percent of linolenic acid ceramide and 0 to 1 weight percent of behenic acid ceramide.

7. A method of synthesis of shea butter ceramide as claimed in any one of claims 1 to 6, comprising the steps of:

under the conditions of a condensing agent and organic base, shea butter fatty acid reacts with a sphingoid compound, wherein the condensing agent is EDCI, and the organic base is DIPEA;

the molar ratio of the shea butter fatty acid to the sphingoid compound to the EDCI to the DIPEA is 1: (1-1.5): (1-2): (1-2), wherein the solvent for reaction is at least one of dichloromethane, tetrahydrofuran, ethyl acetate and acetonitrile.

8. Use of shea butter ceramide as claimed in any one of claims 1 to 6 in cosmetics, pharmaceuticals, dietary foods or health products.

9. The use according to claim 8, wherein the shea butter ceramide has at least one of skin barrier repair, tissue healing, anti-aging, anti-photoaging, anti-oxidant, collagen synthesis promotion, elastin viability maintenance, and whitening efficacy.

10. A composition comprising the shea butter ceramide of any one of claims 1-6, having at least one of skin barrier repair, tissue healing, anti-aging, anti-photoaging, anti-oxidant, collagen synthesis promotion, elastin viability maintenance, and whitening efficacy.