Method for preparing multi-dimensional co-assembled hydroponic oil and application of multi-dimensional co-assembled hydroponic oil

Abstract

The invention provides a method for preparing multi-dimensional co-assembled hydroponic oil and application of the multi-dimensional co-assembled hydroponic oil, and belongs to the technical field of grease fermentation. The method comprises the following steps: S1, taking an oil-insoluble substance, mixing the oil-insoluble substance with soybean phospholipid in absolute ethyl alcohol, stirring at 50-70 DEG C until the solution is clear, centrifuging, taking a supernatant, and volatilizing a solvent to obtain a compound; s2, dissolving chitosan and fermentation oil in an acetic acid solution with the concentration of 0.5-2 wt%, uniformly mixing, and standing to obtain a mixed solution; s3, dissolving the compound obtained in the step S1 in absolute ethyl alcohol, slowly dropwise adding into the mixed solution obtained in the step S2 in a constant-speed stirring state, and stirring until the mixed solution emits micro-blue opalescence to obtain a co-assembled solution; and S4, adding the co-assembled solution in the S3 into grease to obtain the multi-dimensional co-assembled hydroponic oil. According to the method, oil-insoluble components can be wrapped and dissolved in grease, the structure is stable, the skin permeation and absorption performance of effective components can be improved, and the grease is endowed with more dermatological functional activity.

Description

Technical Field

[0001] The present invention relates to the technical field of oil fermentation, and in particular to a method for preparing multi-dimensional co-assembled hydroponic oil and application thereof in cosmetics and / or skin care products. Background Art

[0002] The human skin is mainly divided into the epidermis, dermis and subcutaneous fat, among which the epidermis is further divided into the stratum corneum, the clear layer, the granular layer, the organic layer and the basal layer. The stratum corneum, the outermost layer of the skin, is composed of a layer of keratin cells, and the spaces between the stratum corneum cells are filled with structures such as ceramide, cholesterol and fatty acids.

[0003] At present, in the oil skin care product market, most products exist in the form of crude oil, such as sunflower seed oil, camellia seed oil, peony seed oil, jojoba oil, baobab oil, etc. However, crude oil lacks characteristic activity and cannot play a certain active function while moisturizing the skin, and most active substances cannot exist stably in oil. Therefore, finding a suitable technology to make oil-insoluble substances exist stably in oil and give oil characteristic functional activity has great research value and market prospects. Summary of the invention

[0004] In view of the above problems existing in the prior art, the present invention provides a method for preparing multi-dimensional co-assembled hydroponic oil and its application. The method of the present invention can encapsulate oil-insoluble components, dissolve them in oil, stabilize the structure, and improve the skin penetration and absorption performance of the active ingredients, giving the oil more dermatological functional activity.

[0005] The technical solution of the present invention is as follows: A method for preparing multi-dimensional co-assembled hydroponic oil comprises the following steps: S1: Take the oil-insoluble substance, mix it with soybean lecithin in anhydrous ethanol, stir it at 50-70°C until the solution is clear, centrifuge it, take the supernatant, evaporate the solvent, and obtain the complex; S2: dissolving chitosan and fermented oil in an acetic acid solution having a concentration of 0.5-2 wt%, mixing them evenly, and standing them to obtain a mixed solution; S3: dissolving the complex obtained in step S1 in anhydrous ethanol, slowly adding the mixture dropwise to the mixed solution obtained in step S2 under uniform stirring, and stirring until the mixed solution emits a light blue opalescent light, thereby obtaining a co-assembly solution; S4: Add the co-assembled solution in S3 to the oil to obtain multi-dimensional co-assembled hydroponic oil.

[0006] Preferably, in step S1, the oil-insoluble substance is at least one of glabridin, dihydroquercetin, ginsenoside, resveratrol, paeonol, naringin, hesperidin, tetrahydrocurcumin, quercetin, curcumin, anthocyanidin, proanthocyanidin, tea polyphenols, and arbutin; and the mass ratio of the oil-insoluble substance to soybean lecithin is 1:8-10.

[0007] Furthermore, the amount of anhydrous ethanol in step S1 is not limited.

[0008] Preferably, in step S2, the amount of chitosan added is 1-1.5% of the mass of the acetic acid solution, and the amount of fermented oil added is 1-5% of the mass of the acetic acid solution.

[0009] Furthermore, in step S2, the method for preparing the fermented oil comprises the following steps: M1: Select a strain that can produce surfactant and cultivate it into seed liquid; M2: Inoculate the seed liquid into the fermentation medium for pre-fermentation; M3: Add oil to continue fermentation; M4: After the fermentation is completed, a demulsifier is added to separate the water and oil phases, and the oil phase is collected, which is the fermented oil.

[0010] Preferably, in step M1, the strain capable of producing a surfactant is at least one of Lactobacillus, Bacillus, Saccharomyces, and Corynebacterium glutamicum; The culture medium formula used to culture the strain into seed liquid is: 8-10 g / L glucose, 15-20 g / L rice, and the culture is carried out at 28-30° C. for 12-24 h after inoculation.

[0011] Preferably, in step M2, the fermentation medium formula is: 15-20 g / L glucose, 6-10 g / L yeast extract, 15-20 g / L rice, and the pre-fermentation is cultured at 28-30° C. for 12-24 h.

[0012] Preferably, in step M3 and step S4, the oils and fats include but are not limited to soybean oil, coconut oil, sunflower seed oil, jojoba seed oil, rosehip oil, grape seed oil, almond oil, white meadowfoam seed oil, macadamia nut oil, olive oil, nut seed oil, camellia seed oil, prickle oil, sea buckthorn seed oil, peony seed oil, hazelnut oil, rice germ oil, wheat germ oil, borage seed oil, safflower seed oil, echium seed oil, European blueberry seed oil, baobab seed oil, palm fruit oil, Chilean hazel oil, African wild mango oil, white lupine oil, and olive shell oil.

[0013] Preferably, in step M3, the volume ratio of the oil to the fermentation medium is 1:0.8-1.2; and the fermentation time is 24-36 hours.

[0014] Furthermore, the demulsifier in step M4 is a combination of sodium chloride and hydrochloric acid, wherein the mass of sodium chloride is 1.5-2.5% of the fermentation product, the mass of hydrochloric acid is 0.1-0.5% of the fermentation product, and the concentration of hydrochloric acid is 5-7M.

[0015] Preferably, in step S3, the concentration of the complex after being dissolved in ethanol is 20-50 mg / mL; the mass of the complex is 10-20% of the mass of the mixed solution.

[0016] Preferably, in step S4, the amount of the co-assembly solution added is 1-5% of the mass of the multi-dimensional co-assembly hydroponic oil.

[0017] Furthermore, the fermented oil can also be refined using an integrated membrane system or silica gel.

[0018] The beneficial technical effects of the present invention are: 1. The lactobacillus selected in the present invention can simultaneously produce surfactants and some small molecule acid substances, wherein the surfactant has an emulsifying effect and can promote the generation of the co-assembly process; and the small molecule acid substances can promote the absorption of fat by the human body.

[0019] 2. Compared with traditional encapsulation or co-assembly technology, the co-assembly process of the present invention does not introduce exogenous emulsifiers other than natural oils and fats. Instead, fermented oil prepared by bacterial fermentation is used to replace the emulsifier function, which is more natural, skin-friendly and less irritating.

[0020] 3. Compared with the traditional method of searching for oils with characteristic activity, the present invention uses co-assembly technology to embed oil-insoluble substances, and the obtained co-assembled hydroponic oil has a stronger effect, can artificially give oils characteristic activity, and has lower cost and is more convenient. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a graph showing the blood plate screening results of Example 1; Figure 2 This is a schematic diagram of the red circle for oil drainage in Example 2. DETAILED DESCRIPTION

[0022] The present invention is described in detail below in conjunction with the accompanying drawings and embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] Example 1: Initial screening of surfactant-producing strains In this example, the blood agar plate method is used to initially select surfactant-producing strains, and the specific method is as follows: (1) Weigh 3 g (3 mL) of soil samples (water samples) from Yandang Mountain in Zhejiang, Tianmu Lake in Liyang, Laoshan in Nanjing, and Xingtai in Hebei, add 30 mL of deionized water, and activate at 30°C and 200 rpm for 30 min. Pipette 1 mL of activated soil suspension and activated water suspension into 20 mL of enrichment medium, and place at 30°C and 200 rpm for 2 days.

[0024] The formula of the enrichment medium (1 L) is: 2 g glucose, 0.2 g yeast extract, 0.2 g NH4NO3, 0.2 g KH2PO4, 0.3 g Na2HPO4·12H2O, 0.5 g, and 0.05 g MgSO4·7H2O.

[0025] (2) Take the enriched culture medium after 2 days of culture and dilute it by 10 6 -10 8 times, spread 100 μL on a blood agar plate, and invert it in a constant temperature incubator at 30°C for 24 h.

[0026] (3) Select strains with larger hemolysis zones and clear edges (e.g. Figure 1 As shown in the figure), streak the plate onto a solid LB agar plate and prepare glycerol tubes for storage at -70°C.

[0027] (4) The 7 strains with the best effects were subjected to 16S and ITS assays, and the strains obtained were Lactobacillus, Bacillus subtilis, Bacillus cereus, Bacillus amyloliquefaciens, Corynebacterium glutamicum, Candida albicans, and Pichia pastoris.

[0028] Example 2: Rescreening of surfactant-producing strains The strains initially screened in Example 1 were further screened by the oil-discharging red circle method, and the specific method was as follows: Take a clean glass culture dish (9 cm in diameter) and place it on the coordinate paper. Add 40 mL of deionized water to the culture dish, and add 200 µL of oil red solution after the water surface stabilizes. When the oil red is completely and evenly spread, add 2.5 µL of fermentation liquid supernatant to the center of the oil red surface. When the size of the transparent oil-draining ring stabilizes, record the diameter of the oil-draining ring. The activity of the surfactant produced by fermentation can be judged by observing the diameter of the oil-draining red ring (e.g. Figure 2 The larger the diameter of the red oil-discharging circle, the stronger the activity of the surfactant produced by fermentation.

[0029] The experimental results show that the diameter of the red circle of lactobacillus oil excretion is the largest, indicating that the surfactant produced by its fermentation is the most active.

[0030] Example 3: Oil fermentation The fermented oil was prepared using the lactobacillus strain screened in Example 2, and the specific process was as follows: M1. Inoculate the lactic acid bacteria screened in Example 2 into a seed culture medium with the following formula: 10 g / L glucose and 20 g / L rice. After inoculation, culture at 30° C. for 12 h.

[0031] M2. The seed liquid cultured in step M1 was inoculated into the fermentation medium for pre-fermentation. The medium formula was: 20 g / L glucose, 10 g / L yeast extract, 20 g / L rice, and cultured at 30°C for 12 h.

[0032] M3, add 1 volume of sunflower oil to the fermentation medium, and continue fermentation for 24 h; M4. After the fermentation is finished, a demulsifier is added to separate the water and oil phases, centrifuge, and take the upper oil phase to obtain the fermented oil. The demulsifier is a combination of sodium chloride and hydrochloric acid, wherein the mass of sodium chloride is 2% of the fermentation product, the mass of hydrochloric acid is 0.1% of the fermentation product, and the concentration of hydrochloric acid is 6M.

[0033] Example 4: Preparation of multi-dimensional co-assembled hydroponic oil with whitening characteristics (1) Take 1 g of glabridin and 10 g of soybean lecithin and fully dissolve them in anhydrous ethanol. Stir at 60°C until the solution is clear and free of precipitation. Centrifuge and take the supernatant. Evaporate the solvent to obtain the glabridin complex.

[0034] (2) Take 1 g of chitosan and 5 g of the fermented oil prepared in Example 3, dissolve them in 100 g of 1 wt% acetic acid solution, stir them thoroughly and let them stand to obtain a mixed solution, and take 100 g of it for later use.

[0035] (3) Take 200 mg of the glabridin complex prepared in step (1), dissolve it in 10 ml of anhydrous ethanol, and add it dropwise to 100 g of the mixed solution prepared in step (2) under uniform stirring. Continue stirring until a slightly blue opalescent light appears to obtain a co-assembly solution.

[0036] (4) Take 5 g of the co-assembly solution prepared in step (3) and add it to 95 g of sunflower seed oil to obtain a multi-dimensional co-assembly hydroponic oil with whitening characteristics.

[0037] (5) The ability of multidimensional co-assembled hydroponic oil to inhibit tyrosinase was determined, and sunflower seed crude oil was used as a control. The specific results are shown in Table 1.

[0038] Table 1 Tyrosinase inhibition ability

[0039] It can be seen from Table 1 that the hydroponic oil obtained by the co-assembly technology has a much higher ability to inhibit tyrosinase than sunflower seed crude oil. The present invention endows crude oil with characteristic whitening activity through this technology.

[0040] Example 5: Preparation of multidimensional co-assembled hydroponic oil with antioxidant activity Take 1 g of dihydroquercetin and 8 g of soybean lecithin and fully dissolve them in anhydrous ethanol, stir at 50°C until clear without precipitation, centrifuge, take the supernatant, and evaporate the solvent to obtain a dihydroquercetin complex.

[0041] Take 1.5 g of chitosan and 1 g of the fermented oil prepared in Example 3, dissolve them in 100 g of 1 wt % acetic acid solution, stir them thoroughly and let them stand to obtain a mixed solution, and take 100 g of it for later use.

[0042] Take 750 mg of the dihydroquercetin complex prepared in step (1), dissolve it in 15 ml of anhydrous ethanol, and add it dropwise to 100 g of the mixed solution prepared in step (2) under stirring. Continue stirring until a slightly blue opalescent light appears to obtain a co-assembly solution.

[0043] Take 1 g of the co-assembly solution prepared in step (3) and add it to 99 g of sunflower oil to obtain a multidimensional co-assembly hydroponic oil with antioxidant characteristics.

[0044] The DPPH removal rate of the multi-dimensional co-assembled hydroponic oil was measured, and sunflower seed crude oil was used as a control. The specific results are shown in Table 2.

[0045] Table 2 DPPH scavenging ability

[0046] It can be seen from Table 2 that the hydroponic oil obtained by the co-assembly technology has a much higher DPPH free radical scavenging ability than the sunflower seed crude oil. The present invention endows crude oil with characteristic free radical scavenging activity through this technology.

[0047] Example 6: Preparation of multidimensional co-assembled hydroponic oil with antibacterial activity Take 1 g of ginsenoside and 9 g of soybean lecithin and fully dissolve them in anhydrous ethanol, stir at 50°C until clear without precipitation, centrifuge, take the supernatant, and evaporate the solvent to obtain the ginsenoside complex.

[0048] Take 1.2 g of chitosan and 2.5 g of the fermented oil prepared in Example 3, dissolve them in 100 g of 1.5 wt % acetic acid solution, stir them thoroughly and let them stand to obtain a mixed solution, and take 100 g of it for later use.

[0049] Take 360 ​​mg of the ginsenoside complex prepared in step (1), dissolve it in 12 ml of anhydrous ethanol, and add it dropwise to 100 g of the mixed solution prepared in step (2) under stirring. Continue stirring until a slightly blue opalescent light appears to obtain a co-assembly solution.

[0050] Take 2.5 g of the co-assembly solution prepared in step (3) and add it to 97.5 g of sunflower oil to obtain a multi-dimensional co-assembly hydroponic oil with antibacterial activity characteristics.

[0051] The size of the antibacterial zone of the multi-dimensional co-assembled hydroponic oil was measured, and sunflower seed crude oil was used as a control, and the antibacterial zone diameter was expressed. The specific results are shown in Table 3.

[0052] Table 3 Antibacterial ability test

[0053] It can be seen from Table 3 that the antibacterial ability of the hydroponic oil obtained by the co-assembly technology is much higher than that of the sunflower seed crude oil. The present invention endows the crude oil with characteristic antibacterial activity through this technology.

[0054] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and implementation modes, and can be fully applicable to various fields suitable for the present invention. For those familiar with the art, for those of ordinary skill in the art, various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principle and spirit of the present invention. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to specific details.

Claims

1. A method for preparing multi-dimensional co-assembled hydroponic oil, characterized in that: The following steps are involved: S1: Take the oil-insoluble substance, mix it with soybean lecithin in anhydrous ethanol, stir it at 50-70°C until the solution is clear, centrifuge it, take the supernatant, evaporate the solvent, and obtain the complex; S2: dissolving chitosan and fermented oil in an acetic acid solution having a concentration of 0.5-2 wt%, mixing them evenly, and standing them to obtain a mixed solution; S3: dissolving the complex obtained in step S1 in anhydrous ethanol, slowly adding the mixture dropwise to the mixed solution obtained in step S2 under uniform stirring, and stirring until the mixed solution emits a light blue opalescence, thereby obtaining a co-assembly solution; S4: Add the co-assembled solution in S3 to the oil to obtain multi-dimensional co-assembled hydroponic oil.

2. The method according to claim 1, characterized in that In step S1, the oil-insoluble substance is at least one of glabridin, dihydroquercetin, ginsenoside, resveratrol, paeonol, naringin, hesperidin, tetrahydrocurcumin, quercetin, curcumin, anthocyanidin, proanthocyanidin, tea polyphenols, and arbutin; the mass ratio of the oil-insoluble substance to soybean lecithin is 1:8-10.

3. The method according to claim 1, characterized in that In step S2, the amount of chitosan added is 1-1.5% of the mass of the acetic acid solution, and the amount of fermented oil added is 1-5% of the mass of the acetic acid solution.

4. The method according to claim 1, characterized in that: In step S2, the method for preparing fermented oil comprises the following steps: M1: Select a strain that can produce surfactant and cultivate it into seed liquid; M2: Inoculate the seed liquid into the fermentation medium for pre-fermentation; M3: Add oil to continue fermentation; M4: After the fermentation is completed, a demulsifier is added to separate the water and oil phases, and the oil phase is collected, which is the fermented oil.

5. The method according to claim 4, characterized in that In step M1, the strain capable of producing a surfactant is at least one of lactobacillus, bacillus, yeast, and Corynebacterium glutamicum; The culture medium formula used to culture the strain into seed liquid is: 8-10 g / L glucose, 15-20 g / L rice, and the culture is carried out at 28-30° C. for 12-24 h after inoculation.

6. The method according to claim 4, characterized in that In step M2, the fermentation medium formula is: 15-20 g / L glucose, 6-10 g / L yeast extract, 15-20 g / L rice, and the pre-fermentation is cultured at 28-30° C. for 12-24 h.

7. The method according to claim 1 or 4, characterized in that: In step M3 and step S4, the oils include but are not limited to soybean oil, coconut oil, sunflower seed oil, jojoba seed oil, rosehip oil, grape seed oil, almond oil, white meadowfoam seed oil, macadamia oil, olive oil, nut seed oil, camellia seed oil, prickle oil, sea buckthorn seed oil, peony seed oil, hazelnut oil, rice germ oil, wheat germ oil, borage seed oil, safflower seed oil, echium seed oil, European blueberry seed oil, baobab seed oil, palm fruit oil, Chilean hazel oil, African wild mango oil, white lupine oil, and olive shell oil.

8. The method according to claim 4, characterized in that In step M3, the volume ratio of the oil to the fermentation medium in step M2 is 1:0.8-1.2; the fermentation time is 24-36 hours.

9. The method according to claim 1, characterized in that: In step S3, the concentration of the complex after being dissolved in ethanol is 20-50 mg / mL; the mass of the complex is 10-20% of the mass of the mixed solution.

10. The method according to claim 1, characterized in that In step S4, the amount of the co-assembly solution added is 1-5% of the mass of the multi-dimensional co-assembly hydroponic oil.

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