A method for preparing a multi-dimensional co-assembled hydroponic oil and its application

Abstract

The present invention provides a method for preparing a multi-dimensional co-assembled hydroponic oil and its application, belonging to the technical field of oil fermentation. The method comprises the following steps: S1: Take an oil-insoluble substance and mix it with soybean phospholipid in absolute ethanol, stir at 50-70 °C until the solution is clear, centrifuge, take the supernatant, and evaporate the solvent to obtain a complex; S2: Dissolve chitosan and fermented oil in an acetic acid solution with a concentration of 0.5-2 wt%, mix evenly, and let stand to obtain a mixed solution; S3: Dissolve the complex obtained in step S1 in absolute ethanol, and slowly add it dropwise to the mixed solution obtained in step S2 under uniform stirring until it emits a faint blue opalescence to obtain a co-assembled solution; S4: Add the co-assembled solution in S3 to the oil to obtain a multi-dimensional co-assembled hydroponic oil. The method of the present invention can encapsulate oil-insoluble components, dissolve them in the oil, with a stable structure, and can also improve the skin penetration and absorption performance of the active ingredients, endowing the oil with more dermatological functional activities.

Description

Technical Field

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

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

[0003] In the current market of oil-based skin care products, most products exist in the form of crude oil, such as sunflower oil, camellia seed oil, peony seed oil, jojoba oil, baobab oil, and so on. However, crude oil lacks characteristic activities and cannot play certain active functions while moisturizing the skin, and most active substances cannot stably exist in oils. Therefore, finding a suitable technology to make oil-insoluble substances stably exist in oils and endow oils with characteristic functional activities 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 a multi-dimensional co-assembled hydroponic oil and its application. The method of the present invention can encapsulate oil-insoluble components, dissolve them in oils, with a stable structure, and can also improve the skin penetration and absorption performance of active ingredients, endowing oils with more skin care functional activities.

[0005] The technical solution of the present invention is as follows:

[0006] A method for preparing a multi-dimensional co-assembled hydroponic oil, comprising the following steps:

[0007] S1: Take an oil-insoluble substance, mix it with soybean phospholipid in absolute ethanol, stir at 50-70 °C until the solution is clear, centrifuge, take the supernatant, and evaporate the solvent to obtain a complex;

[0008] S2: Dissolve chitosan and fermented oil in an acetic acid solution with a concentration of 0.5-2 wt%, mix evenly, and let stand to obtain a mixed solution;

[0009] S3: Dissolve the complex obtained in step S1 in absolute ethanol, and slowly add it dropwise to the mixed solution obtained in step S2 under uniform stirring until a faint blue opalescence appears to obtain a co-assembled solution;

[0010] S4: Add the co-assembled solution in S3 to an oil to obtain a multi-dimensional co-assembled hydroponic oil.

[0011] Preferably, in step S1, the oil-insoluble substance is at least one of glabridin, dihydroquercetin, ginsenoside, resveratrol, paeonol, naringin, hesperidin, tetrahydrocurcumin, quercetin, curcumin, anthocyanin, proanthocyanidin, tea polyphenol, arbutin; the mass ratio of the oil-insoluble substance to soybean phospholipid is 1:8-10.

[0012] Furthermore, in step S1, the amount of absolute ethanol used is not limited.

[0013] Preferably, in step S2, the addition amount of chitosan is 1-1.5% of the mass of the acetic acid solution, and the addition amount of the fermented oil is 1-5% of the mass of the acetic acid solution.

[0014] Furthermore, in step S2, the preparation method of the fermented oil includes the following steps:

[0015] M1: Select a strain capable of producing a surfactant and culture it into a seed solution;

[0016] M2: Inoculate the seed solution into a fermentation medium and perform pre-fermentation;

[0017] M3: Add oil for continuous fermentation;

[0018] M4: After the fermentation is completed, add a demulsifier to separate the water and oil phases, and collect the oil phase, which is the fermented oil.

[0019] Preferably, in step M1, the strain capable of producing a surfactant is at least one of Lactobacillus, Bacillus, Saccharomyces, Corynebacterium glutamicum;

[0020] The culture medium formula for culturing the strain into a seed solution is: glucose 8-10 g / L, rice 15-20 g / L, and culture at 28-30 °C for 12-24 h after inoculation.

[0021] Preferably, in step M2, the fermentation medium formula is: glucose 15-20 g / L, yeast extract 6-10 g / L, rice 15-20 g / L, and the pre-fermentation is at 28-30 °C for 12-24 h.

[0022] Preferably, in steps M3 and S4, the oils include but are not limited to soybean oil, coconut oil, sunflower seed oil, jojoba oil, rosehip oil, grape seed oil, almond oil, tamanu oil, macadamia nut oil, olive fruit oil, candlenut seed oil, camellia seed oil, pricklyash seed oil, seabuckthorn seed oil, peony seed oil, hazelnut oil, rice germ oil, wheat germ oil, borage oil, safflower oil, echium oil, bilberry oil, baobab oil, palm fruit oil, Chilean hazelnut oil, African wild mango oil, white lupin oil, olive shell oil.

[0023] Preferably, in step M3, the volume ratio of the oil to the fermentation medium is 1:0.8 - 1.2; the time for continuous fermentation is 24 - 36 h.

[0024] Further, the demulsifier in step M4 is a composition 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 - 7 M.

[0025] Preferably, in step S3, the concentration of the complex dissolved in ethanol is 20 - 50 mg / mL; the mass of the complex is 10 - 20% of the mass of the mixed solution.

[0026] Preferably, in step S4, the addition amount of the co - assembly solution is 1 - 5% of the mass of the multi - dimensional co - assembly hydroponic oil.

[0027] Further, the fermented oil can also be refined using an integrated membrane system or silica gel.

[0028] The beneficial technical effects of the present invention are as follows:

[0029] 1. The selected Lactobacillus in the present invention can simultaneously produce surfactants and some small - molecule acid substances. Among them, surfactin has an emulsifying effect and can promote the formation of the co - assembly process; the small - molecule acids can promote the human body's absorption of oil.

[0030] 2. Compared with traditional embedding or co - assembly techniques, in the co - assembly process of the present invention, no exogenous emulsifier of non - natural oil is introduced. Instead, the fermented oil prepared by strain fermentation is used to replace the emulsifier function, which is more natural and skin - friendly and has less irritation.

[0031] 3. Compared with the traditional method of searching for oils with characteristic activities, the present invention embeds oil - insoluble substances through co - assembly technology, and the obtained co - assembly hydroponic oil has a stronger effect. It can artificially endow the oil with characteristic activities, has a lower use cost, and is more convenient. Description of the Drawings

[0032] Figure 1 It is the blood plate screening result diagram of Example 1;

[0033] Figure 2 It is the schematic diagram of the oil - draining red circle of Example 2. Detailed Embodiments

[0034] The present invention will be specifically described 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 of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.

[0035] Example 1: Primary screening of surfactant-producing strains

[0036] In this example, the blood agar plate method was used for the primary selection of surfactant-producing strains. The specific method is as follows:

[0037] (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 respectively, add 30 mL of deionized water, and activate at 30 °C and 200 rpm for 30 min. Transfer 1 mL of the activated soil suspension and activated water sample suspension respectively and inoculate them into 20 mL of enrichment medium, and culture them on a shaker at 30 °C and 200 rpm for 2 days.

[0038] The formula of the enrichment medium (1 L) is: glucose 2 g, yeast extract 0.2 g, NH4NO3 0.2 g, KH2PO4 0.3 g, Na2HPO4·12H2O 0.5 g, MgSO4·7H2O 0.05 g.

[0039] (2) Take the enrichment culture solution cultured for 2 d, dilute it serially by 10 6 -10 8 times, take 100 μL and spread it on a blood agar plate, and incubate it upside down in a constant temperature incubator at 30 °C for 24 h.

[0040] (3) Select the strains with larger hemolytic zones and clear edges (as Figure 1 shown), streak-inoculate them onto a solid LB agar plate for standby, and prepare glycerol tubes for storage at -70 °C.

[0041] (4) Perform 16S and ITS determinations on the 7 strains with the best screening effects respectively, and the obtained strains are Lactobacillus, Bacillus subtilis, Bacillus cereus, Bacillus amyloliquefaciens, Corynebacterium glutamicum, Candida, and Pichia guilliermondii.

[0042] Example 2: Secondary screening of surfactant-producing strains

[0043] The strains obtained from the primary screening in Example 1 were further subjected to secondary screening by the method of oil-removing red circles. The specific method is as follows:

[0044] Place a clean glass Petri dish (9 cm in diameter) on the coordinate paper. Add 40 mL of deionized water to the Petri dish. After the water surface stabilizes, add 200 µL of oil red solution. After the oil red is completely and evenly spread, add 2.5 µL of the supernatant of the fermentation broth to the center of the oil red surface. Record the diameter of the oil drainage circle when the size of the transparent oil drainage circle stabilizes. The activity of the surfactant produced by fermentation is judged by observing the diameter of the oil red drainage circle (as Figure 2 shown). The larger the diameter of the oil red drainage circle, the stronger the activity of the surfactant produced by fermentation.

[0045] The experimental results show that the diameter of the oil red drainage circle of Lactobacillus is the largest, indicating that the surfactant produced by its fermentation has the strongest activity.

[0046] Example 3: Oil fermentation

[0047] Prepare fermented oil using the Lactobacillus strain screened in Example 2. The specific process is as follows:

[0048] M1. Inoculate the Lactobacillus screened in Example 2 into the seed medium. The medium formula is: 10 g / L of glucose, 20 g / L of rice. After inoculation, culture at 30 °C for 12 h.

[0049] M2. Inoculate the seed liquid cultured in step M1 into the fermentation medium for pre-fermentation. The medium formula is: 20 g / L of glucose, 10 g / L of yeast extract, 20 g / L of rice. Culture at 30 °C for 12 h.

[0050] M3. Add sunflower seed oil with a volume 1 time that of the fermentation medium, and continue fermentation culture for 24 h;

[0051] M4. After fermentation, add a demulsifier to separate the water and oil phases, centrifuge, and take the upper oil phase to obtain fermented oil. The demulsifier is a composition of sodium chloride and hydrochloric acid, where 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 6 M.

[0052] Example 4: Preparation of a multi-dimensional co-assembled hydroponic oil with whitening characteristics

[0053] (1) Take 1 g of glabridin, and fully dissolve it with 10 g of soybean phospholipids in absolute ethanol. Stir at 60 °C until clear and without precipitation, centrifuge, take the supernatant, and evaporate the solvent by rotary evaporation to obtain a glabridin complex.

[0054] (2) Take 1 g of chitosan and 5 g of the fermented oil prepared in Example 3, dissolve them in 100 g of an acetic acid solution with a concentration of 1 wt%, stir well, let stand, obtain a mixed solution, and take 100 g of it for use.

[0055] (3) Take 200 mg of the glabridin complex prepared in step (1), dissolve it in 10 ml of absolute ethanol, and drop it into 100 g of the mixed solution prepared in step (2) under uniform stirring. Keep stirring until a slightly blue opalescence appears to obtain a co-assembly solution.

[0056] (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-assembled hydroponic oil with whitening characteristics.

[0057] (5) Measure the ability of the multi-dimensional co-assembled hydroponic oil to inhibit tyrosinase. At the same time, use crude sunflower seed oil as a control. The specific results are shown in Table 1.

[0058] Table 1 Ability to inhibit tyrosinase

[0059]

[0060] As can be seen from Table 1, the hydroponic oil obtained by the co-assembly technology has a much higher ability to inhibit tyrosinase than crude sunflower seed oil. Through this technology, the present invention endows the crude oil with the characteristic activity of whitening.

[0061] Example 5: Preparation of multi-dimensional co-assembled hydroponic oil with antioxidant activity

[0062] Take 1 g of dihydroquercetin and fully dissolve it with 8 g of soybean phospholipids in absolute ethanol. Stir at 50 °C until clear and without precipitation, centrifuge, take the supernatant, and rotary evaporate to dry the solvent to obtain a dihydroquercetin complex.

[0063] Take 1.5 g of chitosan and 1 g of the fermented oil prepared in Example 3, dissolve them in 100 g of acetic acid solution with a concentration of 1 wt%, stir well, let stand, and obtain a mixed solution. Take 100 g of it for standby.

[0064] Take 750 mg of the dihydroquercetin complex prepared in step (1), dissolve it in 15 ml of absolute ethanol, and drop it into 100 g of the mixed solution prepared in step (2) under stirring. Keep stirring until a slightly blue opalescence appears to obtain a co-assembly solution.

[0065] Take 1 g of the co-assembly solution prepared in step (3) and add it to 99 g of sunflower seed oil to obtain a multi-dimensional co-assembled hydroponic oil with antioxidant characteristics.

[0066] Measure the DPPH scavenging rate of the multi-dimensional co-assembled hydroponic oil. At the same time, use crude sunflower seed oil as a control. The specific results are shown in Table 2.

[0067] Table 2 DPPH scavenging ability

[0068]

[0069] As can be seen from Table 2, the DPPH free radical scavenging ability of the hydroponic oil obtained by the co-assembly technology is much higher than that of the crude sunflower oil. The present invention endows the crude oil with the characteristic activity of scavenging free radicals through this technology.

[0070] Example 6: Preparation of multi-dimensional co-assembled hydroponic oil with antibacterial activity

[0071] Take 1 g of ginsenoside, dissolve it fully with 9 g of soybean phospholipid in absolute ethanol, stir at 50 °C until clear and without precipitation, centrifuge, take the supernatant, and evaporate the solvent by rotary evaporation to obtain the ginsenoside complex.

[0072] Take 1.2 g of chitosan and 2.5 g of the fermented oil prepared in Example 3, dissolve them in 100 g of acetic acid solution with a concentration of 1.5 wt%, stir well, let it stand, obtain a mixed solution, and take 100 g of it for standby.

[0073] Take 360 mg of the ginsenoside complex prepared in step (1), dissolve it in 12 ml of absolute ethanol, and drop it into 100 g of the mixed solution prepared in step (2) under stirring, and continue to stir until a faint blue opalescence appears to obtain a co-assembled solution.

[0074] Take 2.5 g of the co-assembled solution prepared in step (3), add it to 97.5 g of sunflower oil to obtain a multi-dimensional co-assembled hydroponic oil with antibacterial activity characteristics.

[0075] Measure the size of the antibacterial zone of the multi-dimensional co-assembled hydroponic oil, and at the same time use the crude sunflower oil as a control, which is represented by the diameter of the antibacterial zone. The specific results are shown in Table 3.

[0076] Table 3 Antibacterial ability test

[0077]

[0078] As can be seen from Table 3, the antibacterial ability of the hydroponic oil obtained by the co-assembly technology is much higher than that of the crude sunflower oil. The present invention endows the crude oil with the characteristic activity of antibacterial through this technology.

[0079] Although the embodiments of the present invention have been disclosed as above, they are not limited to only the applications listed in the specification and embodiments. It can be fully applied to various fields suitable for the present invention. For those skilled in 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 principles and spirits of the present invention. Therefore, without departing from the general concept defined by the claims and the equivalent scope, 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: taking an oil-insoluble substance, mixing it with soybean lecithin in anhydrous ethanol, stirring at 50-70° C. until the solution is clear, centrifuging, taking the supernatant, and evaporating the solvent to obtain a complex; the mass ratio of the oil-insoluble substance to the soybean lecithin is 1:8-10; S2: dissolving chitosan and fermented oil in an acetic acid solution with a concentration of 0.5-2wt%, mixing evenly, and standing 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: adding the co-assembly solution in S3 to the oil to obtain multi-dimensional co-assembly hydroponic oil; 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; 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; In step M1, the strain capable of producing a surfactant is a lactobacillus.

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.

3. The method according to claim 1, characterized in that In step M1, 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 hours after inoculation.

4. The method according to claim 1, 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 hours.

5. The method according to claim 1, 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.

6. The method according to claim 1, 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.

7. 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.

8. 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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