Application of raw material obtained by photolysis of aloe in cosmetics

Abstract

The invention discloses an application of a raw material obtained by performing ultraviolet photolysis on aloe in cosmetics. According to the preparation method, gel and leaf skin in aloe leaves are used as raw materials at the same time, hydrogen peroxide is added as an oxidizing agent, and hydrogen peroxide is decomposed into hydroxyl radicals through irradiation of ultraviolet light; on one hand, the hydroxyl free radicals decompose plant cell walls in aloe leaf skins and release active substances in the aloe leaf skins; meanwhile, plant cellulose in leaves and skins is degraded into small-molecular polysaccharides, some of the small-molecular polysaccharides are further oxidized into small-molecular organic acid, and the small-molecular organic acid is a component which is not contained in a traditional aloe extract; on the other hand, macromolecular polysaccharide in the aloe gel is also degraded into micromolecular polysaccharide and micromolecular organic acid, wherein the components of the micromolecular polysaccharide and the micromolecular organic acid are more easily absorbed by the skin. According to the preparation method, full-component utilization of gel and skin in aloe leaves is realized, generation of waste is reduced, and meanwhile, the raw material obtained by performing ultraviolet photolysis on the aloe has excellent moisturizing, anti-inflammatory and exfoliating effects.

Description

Technical Field

[0001] The invention belongs to the field of cosmetic chemistry and biotechnology, and relates to application of a raw material obtained by ultraviolet photolysis of aloe vera in cosmetics. Background Art

[0002] Aloe vera extract is widely used in the cosmetics field as a raw material for cosmetics or skin care products. Its main function is moisturizing. There are many extraction processes for aloe vera, including extraction, filtration, ultrasound and other methods. The object of traditional extraction process is the viscous liquid in the aloe vera leaves, which is called aloe vera gel. The separated outer leaf skin is often discarded as waste. Very few manufacturers will extract the active substances in the outer leaves through complex and high-cost processes, but the utilization rate is very low, and more solid waste and wastewater are generated. Summary of the invention

[0003] To solve the above problems, the purpose of the present invention is to provide a raw material obtained by ultraviolet photolysis of aloe vera for use in cosmetics. The present invention treats aloe vera by ultraviolet photolysis for the first time to obtain an aloe vera photolysis product.

[0004] The objective of the present invention is achieved through the following technical solutions:

[0005] The invention provides a method for preparing an aloe photolysis product, which comprises subjecting at least one of aloe leaf powder and aloe gel to a photolysis reaction in the presence of hydrogen peroxide to prepare the aloe photolysis product.

[0006] As an embodiment, the photolysis uses ultraviolet light in the 225-365nm band and reacts at 20-30°C.

[0007] As an embodiment, the reaction time of the photolysis reaction is 5-20h.

[0008] As an embodiment, the mass concentration of the hydrogen peroxide is 20-50%.

[0009] As an embodiment, the aloe is selected from one or more of Aloe vera, Aloe antonii, Aloe vera, Aloe arborescens, Aloe vera, and Aloe arborescens.

[0010] As an embodiment, at least one of aloe leaf powder and aloe gel is pretreated with hydrogen peroxide and then subjected to photolysis reaction.

[0011] As an embodiment, the amount of 35 wt. % hydrogen peroxide added to the aloe leaf powder is 10-30% of the weight of the aloe leaf powder.

[0012] As an embodiment, the amount of 35 wt. % hydrogen peroxide added to the aloe gel is 0.5-2% of the weight of the aloe gel.

[0013] As an implementation scheme, aloe leaf powder and aloe gel are pretreated with hydrogen peroxide respectively and then mixed to perform a photolysis reaction.

[0014] As an embodiment, the weight ratio of the aloe leaf powder to the aloe gel is 1:1.5-1:2.5.

[0015] As an embodiment, the aloe leaf powder is obtained by crushing the aloe leaf peel. In some implementation examples, after washing the aloe leaf with clean water, the aloe leaf peel and the aloe gel in the middle are separated with a scraper; and the aloe leaf peel is put into a blender and fully crushed.

[0016] As an embodiment, after the photolysis reaction is completed, a post-decolorization treatment step is also included, such as activated carbon decolorization treatment.

[0017] The aloe photolysis product prepared by the method of the present invention and its application in pharmaceutical products or cosmetics belong to the protection scope of the present invention.

[0018] The present invention also provides a cosmetic comprising the aloe photolysis product prepared by the method of the present invention.

[0019] As an embodiment, the cosmetic is a cosmetic having at least one of moisturizing, anti-inflammatory, and exfoliating effects.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] 1) The present invention treats Aloe vera by ultraviolet photolysis for the first time to obtain an Aloe vera photolysis product. Compared with the common Aloe vera extract on the market, this photolysis Aloe vera product shows different effects in cosmetic efficacy tests. The specific reason is that: in the present invention, the gel and leaf skin in the aloe leaf are used as raw materials at the same time, and hydrogen peroxide is added as an oxidant, and the hydrogen peroxide is decomposed into hydroxyl radicals by ultraviolet light irradiation; on the one hand, the hydroxyl radicals decompose the plant cell walls in the aloe leaf skin and release the active substances therein, such as aloin A, aloin B, aloe-emodin, etc.; at the same time, the plant cellulose in the leaf skin is degraded into small molecular polysaccharides, some of which are further oxidized into small molecular organic acids; on the other hand, the macromolecular polysaccharides in the aloe gel are also degraded into small molecular polysaccharides and small molecular organic acids whose components are more easily absorbed by the skin. The small molecular polysaccharides obtained by the degradation of leaf plant cellulose are more easily absorbed by the skin than macromolecular polysaccharides, producing a better moisturizing effect; the small molecular organic acids generated by further oxidation are components not contained in traditional aloe extracts. These small molecule organic acids can have similar effects to fruit acids, such as exfoliation or peeling.

[0022] 2) The present invention realizes the full utilization of gel and skin in aloe leaves, which not only improves the efficacy but also reduces the generation of waste. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Other features, objects and advantages of the present invention will become more apparent from the detailed description of non-limiting embodiments made with reference to the following drawings:

[0024] Figure 1 This is the liquid chromatogram of the sample before photolysis of aloe;

[0025] Figure 2 is the liquid chromatogram of the aloe sample after photolysis reaction;

[0026] Figure 3 It is a comparison chart of TNF-α content in each experimental group;

[0027] Figure 4 It is a comparison chart of IL-6 content in each experimental group;

[0028] Figure 5 The results of AQP3 immunofluorescence staining in each experimental group;

[0029] Figure 6 It is a comparison chart of relative IOD values ​​of each experimental group;

[0030] Figure 7 This is a comparison chart of BCA protein concentration content detection in each experimental group. DETAILED DESCRIPTION

[0031] The present invention is described in detail below in conjunction with embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that, for those of ordinary skill in the art, some adjustments and improvements can also be made without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0032] Example 1

[0033] After rinsing the aloe vera leaves with clean water, separate the aloe vera leaf peel from the aloe vera gel in the middle with a scraper, wherein the weight ratio of the aloe vera leaf peel to the aloe vera gel is about 1:2. Put the aloe vera leaf peel into a blender and crush it thoroughly. Add different amounts of hydrogen peroxide to the crushed aloe vera leaf peel and aloe vera gel respectively for pretreatment by fully mixing; the amount of 35% hydrogen peroxide added to the aloe vera leaf peel is 20% of the weight of the aloe vera leaf peel, and the amount of 35% hydrogen peroxide added to the aloe vera gel is 1% of the weight of the aloe vera gel. After the pretreatment is completed, the hydrogen peroxide mixture of the aloe vera leaf peel and the aloe vera gel is added to the ultraviolet photolysis reactor for photolysis reaction. The photolysis uses ultraviolet light in the 254nm band, and reacts at 25°C in a kettle or tubular reactor for 8 hours. After the photolysis reaction is completed, the aloe vera photolysis essence can be obtained after decolorization post-treatment with activated carbon.

[0034] The sample liquid chromatogram before photolysis of aloe vera in this example is as follows Figure 1 It contains trace amounts of citric acid, fruit acid and glucose, about 100-200 ppm. The liquid chromatogram of the aloe sample after photolysis is shown in Figure 2 As shown in Table 1, the contents of citric acid, glucose and fructose increased significantly. In addition, glycolic acid produced by photolysis of cellulose and polysaccharides was also detected by HPLC; its content is shown in Table 1.

[0035] Example 2

[0036] After rinsing the aloe vera leaves with clean water, separate the aloe vera leaf peel from the aloe vera gel in the middle with a scraper, wherein the weight ratio of the aloe vera leaf peel to the aloe vera gel is about 1:2. Put the aloe vera leaf peel into a blender and crush it thoroughly. Add different amounts of hydrogen peroxide to the crushed aloe vera leaf peel and aloe vera gel respectively for pretreatment by fully mixing; the amount of 35% hydrogen peroxide added to the aloe vera leaf peel is 10% of the weight of the aloe vera leaf peel, and the amount of 35% hydrogen peroxide added to the aloe vera gel is 2% of the weight of the aloe vera gel. After the pretreatment is completed, the hydrogen peroxide mixture of the aloe vera leaf peel and the aloe vera gel is added to the ultraviolet photolysis reactor for photolysis reaction. The photolysis uses ultraviolet light in the 225nm band, and reacts at 20°C in a kettle or tubular reactor for 10 hours. After the photolysis reaction is completed, the aloe vera photolysis essence can be obtained after decolorization post-treatment with activated carbon. The contents of citric acid, glucose, fructose and glycolic acid in the aloe vera photolysis essence are shown in Table 1.

[0037] Example 3

[0038] After rinsing the aloe vera leaves with clean water, separate the aloe vera leaf peel from the aloe vera gel in the middle with a scraper, wherein the weight ratio of the aloe vera leaf peel to the aloe vera gel is about 1:2. Put the aloe vera leaf peel into a blender and crush it thoroughly. Add different amounts of hydrogen peroxide to the crushed aloe vera leaf peel and aloe vera gel respectively for pretreatment by fully mixing; the amount of 35% hydrogen peroxide added to the aloe vera leaf peel is 30% of the weight of the aloe vera leaf peel, and the amount of 35% hydrogen peroxide added to the aloe vera gel is 0.5% of the weight of the aloe vera gel. After the pretreatment is completed, the hydrogen peroxide mixture of the aloe vera leaf peel and the aloe vera gel is added to the ultraviolet photolysis reactor for photolysis reaction. The photolysis uses ultraviolet light of 365nm band, and reacts at 30℃ in a kettle or tubular reactor for 15h. After the photolysis reaction is completed, the aloe vera photolysis essence can be obtained after decolorization post-treatment with activated carbon. The contents of citric acid, glucose, fructose and glycolic acid in the aloe vera photolysis essence are shown in Table 1.

[0039] Table 1

[0040] Element glucose(%) fructose(%) Citric acid(%) Glycolic acid (%) Example 1 0.39 0.28 0.18 0.2 Example 2 0.33 0.22 0.11 0.13 Example 3 0.32 0.23 0.11 0.12 0.3-0.4 0.2-0.3 0.1-0.2 0.1-0.2

[0041] Efficacy test case

[0042] 1. Anti-inflammatory test

[0043] The test item is "Soothing efficacy test - detection of inflammatory factors (TNF-α, IL-6) content in LPS-stimulated macrophage RAW264.7". The experimental design is as shown in Table 2:

[0044] Table 2

[0045]

[0046]

[0047] Note: Aloe vera photolysis essence sample is prepared by adding water to the aloe vera photolysis essence prepared in Example 1; BC represents cultured cells without any sample added and any treatment; NC represents cultured cells that have been negatively treated and without any sample added; Aloe vera stock solution refers to the extract of aloe vera leaf peel and gel that has not undergone any treatment or chemical reaction, that is, the juice in the crushed aloe vera leaf peel and aloe vera gel (same as in Example 1) is extracted by pure physical means (including crushing, squeezing, filtering, etc.).

[0048] First, macrophages were cultured according to the following steps:

[0049] 1) Cell seeding: Cells were seeded into a 24-well plate at a seeding density of 1×10 5 cells / well and incubated overnight in an incubator (37° C., 5% CO 2 ).

[0050] 2) Solution preparation: Prepare working solutions of the test substance of different concentrations according to the experimental design table.

[0051] 3) Induction and drug administration: When the cell plating rate in the 24-well plate reaches 40% to 60%, add 100 μL of 10×LPS working solution to the well plate according to the experimental design, shake the well plate left and right to mix the drugs in the well plate, and add the sample at the same time. The final concentration of LPS is 1 μg / mL, and each group has 3 replicate wells. After drug administration, place the well plate in an incubator (37°C, 5% CO 2 )Continue culturing for 24 h.

[0052] 4) Sample collection: After the incubation is completed, the cell culture supernatant is collected in an EP tube (Note: the amount of collected samples is determined according to the detection indicators). After collection, the samples are placed in a -80°C refrigerator for freezing and storage.

[0053] Then the contents of TNF-α and IL-6 were detected respectively.

[0054] 1) TNF-α content test results

[0055] Based on the above experimental method, macrophage supernatant was collected and TNF-α content was detected. The results are shown in Table 3:

[0056] Table 3

[0057]

[0058]

[0059] Note: When using the t-test two-tailed test method for statistical analysis, the significance of the NC group compared with the BC group is indicated by #, p-value < 0.05 is indicated by #, and p-value < 0.01 is indicated by ##. The significance of the sample group and PC group compared with the NC group is indicated by *, p-value < 0.05 is indicated by *, and p-value < 0.01 is indicated by **.

[0060] Figure 3 is a comparison chart of TNF-α content in each experimental group; Figure 3It can be seen that compared with the BC group, the secretion of TNF-α, an inflammatory factor, by macrophages RAW264.7 in the NC group increased significantly (p<0.05). Compared with the NC group, the secretion of TNF-α, an inflammatory factor, by macrophages RAW264.7 in the PC group was significantly reduced at a dexamethasone concentration of 100 μg / mL (p<0.01), indicating that the positive control test was effective. Compared with the NC group, the secretion of TNF-α, an inflammatory factor by macrophages RAW264.7, was significantly reduced when the aloe vera stock solution sample was at a concentration of 1.25% (V / V) and the aloe vera photolysis essence sample was at a concentration of 0.31%, 0.63% and 1.25% (V / V) (p<0.05).

[0061] 2) IL-6 content test results

[0062] Based on the above experimental method, macrophage supernatant was collected and IL-6 content was detected. The results are shown in Table 4:

[0063] Table 4

[0064] Group Average concentration (pg / mL) SD p-value Blank control (BC) 90.96 6.85 / Negative control (NC) 139.26 2.46 0.005## Positive Control (PC) 94.43 4.07 0.004** Aloe vera stock solution - 1.25% 96.78 3.56 0.001** Aloe Photolysis Extract - 0.31% 108.21 2.21 0.000* Aloe Photolysis Essence - 0.63% 97.36 1.13 0.000** Aloe Photolysis Essence - 1.25% 93.08 4.45 0.001**

[0065] Note: When using the t-test two-tailed test method for statistical analysis, the significance of the NC group compared with the BC group is indicated by #, p-value < 0.05 is indicated by #, and p-value < 0.01 is indicated by ##. The significance of the sample group and PC group compared with the NC group is indicated by *, p-value < 0.05 is indicated by *, and p-value < 0.01 is indicated by **.

[0066] Figure 4 is a comparison chart of IL-6 content in each experimental group; Figure 4 It can be seen that compared with the BC group, the secretion of IL-6, an inflammatory factor, by macrophages RAW264.7 in the NC group increased significantly (p<0.01), indicating that the LPS stimulation modeling was successful in this experiment. Compared with the NC group, the secretion of IL-6, an inflammatory factor by macrophages RAW264.7 in the PC group was significantly reduced at a dexamethasone concentration of 100 μg / mL (p<0.01), indicating that the positive control test was effective. Compared with the NC group, the secretion of IL-6, an inflammatory factor by macrophages RAW264.7, was significantly reduced when the aloe vera stock solution sample was at a concentration of 1.25% (V / V) and the aloe vera photolysis essence sample was at a concentration of 0.31%, 0.63% and 1.25% (V / V) (p<0.01).

[0067] According to the above test results, based on the LPS-stimulated macrophage RAW264.7 model, the aloe vera photolysis essence sample at concentrations of 0.31%, 0.63% and 1.25% (V / V) significantly reduced the secretion of inflammatory factors TNF-α and IL-6 in macrophage RAW264.7, and there was a statistical difference compared with the NC group (p<0.05), indicating that the aloe vera photolysis essence sample can inhibit the secretion of inflammatory factors TNF-α and IL-6 in macrophage RAW264.7 and has a soothing effect. In addition, compared with the aloe vera gel solution, at the same concentration, the aloe vera photolysis sample has a slightly better effect.

[0068] 2. Moisturizing test

[0069] The test item is the moisturizing effect test - keratinocyte hydrated protein (AQP3) content. The experimental design is as shown in Table 5:

[0070] Table 5

[0071]

[0072] The test steps are as follows:

[0073] 1) Cell seeding: Cells were seeded into a 24-well plate at a seeding density of 1×10 5 cells / well and incubated overnight in an incubator (37° C., 5% CO 2 ).

[0074] 2) Solution preparation: Use HaCaT cell culture medium to prepare three concentrations of the test substance working solutions according to the concentrations listed in the experimental design (Table 3).

[0075] 3) Drug administration: According to the experimental design table, when the cell plating rate in the 24-well plate reaches 40% to 60%, the original cell culture solution is aspirated and then the drug administration is carried out in groups. In the experimental group, 1.0 mL of sample working solution is added to each well, and 3 replicate wells are set for each concentration of working solution. In the control group (BC group), 1.0 mL of cell culture solution is added to each well, and 3 replicate wells are set. Place in the incubator (37°C, 5% CO2) and continue to culture for 24 hours.

[0076] 4) Sample collection: discard the supernatant and rinse the cells 3 times with PBS.

[0077] 5) Immunofluorescence staining: Perform conventional immunofluorescence staining process. The main steps are: fixation, blocking, adding primary antibody, adding secondary antibody, DAPI counterstaining, and then taking pictures using a fluorescence microscope.

[0078] 6) Result analysis: The fluorescence intensity of AQP3 was quantitatively analyzed using ImageProPlus software.

[0079] The results of AQP3 immunofluorescence staining of the samples are as follows Figure 5As shown in Table 6 (green fluorescence represents AQP3, blue fluorescence represents the cell nucleus), the relative IOD values ​​are shown in Table 6 and Figure 6 shown.

[0080] Table 6

[0081] Sample name Relative IOD average SD p-value Blank control (BC) 1.00 0.02 / Aloe vera stock solution - 0.313% 1.09 0.05 0.012 Aloe Photolysis Essence - 0.078% 1.02 0.07 0.660 Aloe Photolysis Extract - 0.157% 1.08 0.03 0.016* Aloe Photolysis Essence - 0.313% 1.10 0.00 0.007**

[0082] Note: When using the t-test two-tailed test method for statistical analysis, the significance of the sample group compared with the BC group is indicated by *, p-value < 0.05 is indicated by *, and p-value < 0.01 is indicated by **.

[0083] Compared with the control BC, the fluorescence intensity of AQP3 protein was significantly enhanced in the sample aloe vera stock solution at a concentration of 0.313%, and in the sample aloe vera photolysis essence at a concentration of 0.157% and 0.313% (V / V) (p < 0.05), indicating that the sample aloe vera photolysis essence can promote the expression of AQP3 in keratinocytes. In addition, compared with the aloe vera gel stock solution, at the same concentration, the aloe vera photolysis sample has a slightly better effect.

[0084] 3. Exfoliation test

[0085] The test items are in vitro exfoliation efficacy - exfoliated keratin total protein BCA method and cell counting method. The experimental design is as shown in Table 7:

[0086] Table 7

[0087]

[0088]

[0089] The test steps are as follows:

[0090] 1) Model preparation: The ex vivo suckling pig skin was taken out from the -20°C freezing condition, placed in 0.9% saline for 30 min of resuscitation, and then the pig skin was taken out with tweezers and dried at room temperature;

[0091] 2) Grouped sampling: According to the experimental design, take equal amounts of samples and evenly apply them on the surface of pig skin and leave them for 10 minutes;

[0092] 3) Patch sampling: Place the D-Squame stratum corneum patch on the surface of the pig skin. The sampler holds the pressure device and presses one end of the pressure device against the patch with a fixed pressure. The patch is applied for 30 seconds under the fixed pressure. After the patch is removed, it is clamped into a 1.5 mL centrifuge tube without contamination.

[0093] 4) Extraction of detached keratin: add 1.5 mL of saline to soak the stratum corneum patch, and then ultrasonicate for 15 min;

[0094] 5) BCA protein concentration test: Take the keratinocyte extracts shed from the stratum corneum patches of each group after ultrasound, and perform quantitative test of BCA protein concentration of stratum corneum shed protein according to the instructions of the kit;

[0095] The test results are as follows:

[0096] Based on the above experimental method, the detached keratin extract was collected and the BCA protein concentration was tested. The test results are shown in Table 8. The change trend is as follows Figure 7 shown.

[0097] Table 8

[0098]

[0099] Note: When the t-test method was used for statistical analysis, the significance of the PC group and the sample group compared with the BC group was indicated by *, p-value < 0.05 was indicated by *, and p-value < 0.01 was indicated by **.

[0100] Compared with the BC group, the protein concentration in the PC group increased significantly (p < 0.01), indicating that the positive control test was effective.

[0101] Compared with the BC group, the protein content of the sample aloe vera photolysis essence increased significantly (p < 0.01), indicating that the sample has an exfoliating effect. However, the sample aloe vera stock solution, at the same concentration, did not have a significant increase in protein content, indicating that it did not have an exfoliating effect.

Claims

1. A method for preparing an aloe photolysis product, characterized in that: At least one of aloe leaf powder and aloe gel is subjected to photolysis reaction in the presence of hydrogen peroxide to obtain the aloe photolysis product.

2. The preparation method according to claim 1, characterized in that: Include at least one of the following technical features: A1. The photolysis is carried out using ultraviolet light in the 225-365nm band at 20-30°C; A2, the mass concentration of hydrogen peroxide is 20-50%; A3, the aloe is selected from one or more of Aloe vera, Aloe antonii, Aloe vera, Aloe arborescens, and Aloe arborescens; A4. The reaction time of the photolysis reaction is 5-20h.

3. The preparation method according to claim 1, characterized in that: The amount of 35wt.% hydrogen peroxide added to the aloe leaf powder is 10-30% of the weight of the aloe leaf powder; the amount of 35wt.% hydrogen peroxide added to the aloe gel is 0.5-2% of the weight of the aloe gel.

4. The preparation method according to claim 1 or 3, characterized in that: Aloe leaf powder and aloe gel were pretreated with hydrogen peroxide respectively and then mixed to carry out photolysis reaction.

5. The preparation method according to claim 4, characterized in that: The weight ratio of the aloe leaf powder to the aloe gel is 1:1.5-1:2.

5.

6. The preparation method according to claim 1, characterized in that: After the photolysis reaction is completed, a decolorization post-treatment step is also included.

7. An aloe photolysis product prepared by the method according to any one of claims 1 to 6.

8. Use of the aloe photolysis product according to claim 7 in pharmaceutical products or cosmetics.

9. A cosmetic comprising the aloe photolysis product prepared by the method according to any one of claims 1 to 6.

10. The cosmetic according to claim 9, characterized in that: The cosmetics are cosmetics having at least one of the functions of moisturizing, anti-inflammatory and exfoliating.

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