Preparation method of lotus seed shell extract with anti-ultraviolet function, obtained product and application

Abstract

The invention belongs to the field of active ingredient extraction, and particularly relates to a preparation method of a lotus seed shell extract with an anti-ultraviolet function, an obtained product and application. The method comprises the following steps: inoculating lotus seed shell powder with a fermentation inoculant and betaine for fermentation; sterilizing and centrifuging the fermentation liquor, and respectively collecting supernate and precipitate; and carrying out ultrasonic extraction on the precipitate by adopting absolute ethyl alcohol, and concentrating to obtain the lotus seed shell ethanol extract. The extraction method provided by the invention is high in extraction efficiency, the free radical scavenging effect of the extract can be improved through fermentation treatment, and the extraction efficiency is high; the method can reduce the damage to compounds in the extraction process, the content of the extracted active ingredients is high, and the development of the biological functions of the lotus seeds is facilitated; according to the ultraviolet protection combined product prepared by the invention, through an internal and external combined protection mode, the external barrier of the skin to ultraviolet rays is improved, the internal antioxidant capacity of the skin is enhanced through oral administration, and comprehensive protection on the skin is realized.

Description

Technical Field

[0001] The invention belongs to the field of active ingredient extraction, and specifically relates to a preparation method of a lotus seed shell extract with anti-ultraviolet function, and the obtained product and application. Background Art

[0002] Ultraviolet radiation is the main cause of skin problems such as skin aging, pigmentation, and skin cancer. Sunscreen products currently on the market mainly form a protective layer on the skin surface through chemical or physical means, but these products often ignore the regulation of internal physiological mechanisms. In addition, many sunscreen ingredients have potential health risks and may have an impact on the environment. Chemical sunscreens widely used in existing sunscreen products, such as oxybenzone and avobenzone, may cause skin irritation or allergic reactions, and are even suspected of having endocrine disrupting effects. In addition, the accumulation of chemical sunscreens in water bodies may have a negative impact on aquatic ecosystems. Most products only provide protection on the skin surface, and do not consider enhancing the skin's natural protective ability through internal regulation, such as improving the skin's antioxidant capacity. They fail to meet the personalized needs of users with different skin types, living habits and environmental factors. Existing sunscreen products rarely consider skin nutrition and repair, and mainly focus on blocking ultraviolet rays rather than comprehensively improving skin health.

[0003] At present, there are many public patents designed and researched for sun protection, but there are still cases where the existing sunscreen product technology is insufficient, which exposes some common problems faced by sunscreen products on the current market. CN202410768850.X proposes a sunscreen composition, but it still relies on chemical sunscreens and may have problems with skin irritation and environmental impact. CN202211517793.5 describes a sunscreen lotion that lacks sufficient skin absorption and utilization efficiency, resulting in poor actual sunscreen effect. In addition, there are public patents for sunscreen products in various forms such as sunscreen spray, sunscreen gel, sunscreen oil, sunscreen stick, sunscreen foundation, sunscreen repair mask, etc. These technologies mainly focus on the innovation of product form, and there are still deficiencies in uniformity, usage experience, skin adaptability, convenience and repair ability.

[0004] In summary, the development of sunscreen products should reduce the use of chemical sunscreens to reduce the risk of skin irritation and allergic reactions; raw materials with high biodegradability and environmental compatibility should be used to reduce the negative impact on the ecosystem. Product formulas should be optimized to improve the skin permeability of active ingredients to ensure that the active ingredients can be fully absorbed and utilized by the skin. Antioxidants, moisturizers and repair ingredients should be combined to provide comprehensive care products that not only protect against the sun, but also nourish and repair the skin.

[0005] Lotus seed shell is the seed shell of lotus, which is traditionally used as a herbal medicine and food in Asia. In recent years, with the deepening of scientific research, people's understanding of lotus seed shell and its active ingredients has continued to deepen. A variety of active ingredients have been identified in lotus seed shell, including polyphenols, flavonoids, alkaloids and polysaccharides. These ingredients give lotus seed shell a variety of biological activities. Lotus seed shell extract exhibits significant antioxidant capacity due to its rich polyphenols, which can scavenge free radicals and reduce oxidative stress. Studies have shown that the active ingredients in lotus seed shell can inhibit the release of inflammatory mediators, have anti-inflammatory effects, and may be beneficial for the treatment of inflammatory diseases. CN202410541497.1 discloses a method for separating and purifying proanthocyanidins in lotus seed shells, but the adaptability of the application of proanthocyanidins is not fully elaborated. CN202311473866.X discloses a lotus seed shell polyphenol extract with hypoglycemic effect and its preparation method and application. In the prior art, there are few reports on lotus seed shell extracts. Summary of the invention

[0006] In view of the problems existing in the prior art, the present invention provides a method for preparing a lotus seed shell extract with anti-ultraviolet function.

[0007] The invention also provides lotus seed shell extract obtained by the preparation method.

[0008] Another object of the present invention is to provide the use of the lotus seed shell extract in the preparation of UV protection products, which can be used in cosmetics as a natural UV filter.

[0009] The technical solution adopted by the present invention to achieve the above-mentioned purpose is: The present invention provides a method for preparing a lotus seed shell extract with anti-ultraviolet function, comprising the following steps: (1) Fermentation treatment: Mix lotus seed shell powder with water, inoculate fermentation agent, add a small amount of betaine, stir evenly and ferment to obtain fermentation liquid; (2) Post-fermentation treatment: sterilize and cool the fermentation broth, then homogenize and centrifuge it, collect the supernatant and precipitate respectively; dry the precipitate for later use; (3) Precipitation extraction: Add 8-10 times the mass of anhydrous ethanol to the dried precipitate and perform ultrasonic extraction. After the extraction, concentrate the precipitate to obtain the lotus seed shell ethanol extract.

[0010] Furthermore, in step (1), the mass ratio of the lotus seed shell powder to water is 1:10-1:15; the fermentation agent is composed of Aspergillus oryzae and Bacillus pumilus in a mass ratio of 2:1; the inoculation amount of the fermentation agent accounts for 2%-3% of the total amount of the lotus seed shell powder; and the added amount of betaine accounts for 0.1%-0.2% of the total amount of the lotus seed shell powder.

[0011] Furthermore, in step (1), the fermentation is carried out at a temperature of 28-30° C. for 60-72 hours.

[0012] Furthermore, in step (2), the homogenization speed is 10000-12000 rpm for 1 min, which is repeated 5-6 times; the centrifugation speed is 6000-8000 rpm, and the centrifugation time is 30 min.

[0013] Furthermore, in step (3), the mass ratio of the precipitate to anhydrous ethanol is 1:8-10; the conditions of the ultrasonic extraction are: ultrasonic frequency 40 kHz, power 100 W, extraction time 30 min, extraction temperature is room temperature; and the extraction is concentrated to 1 / 7~1 / 8 of the original volume.

[0014] The present invention also provides an application of the lotus seed shell extract prepared by the preparation method in preparing an anti-ultraviolet product. The anti-ultraviolet product comprises a beverage containing a supernatant and an emulsion containing the lotus seed shell ethanol extract.

[0015] The beverage containing supernatant provided by the present invention is prepared by the following method: adding the supernatant to purified water in a ratio of 2:1 to obtain a mixed solution; then adding 0.5% ascorbic acid, 0.01% sodium selenate, 0.05% zinc sulfate and 1% stevioside accounting for the total weight of the mixed solution, mixing evenly, filtering, homogenizing, degassing and sterilizing to obtain the beverage.

[0016] The emulsion containing the lotus seed shell ethanol extract provided by the present invention is prepared by the following method: (a) Preparation of emulsion oil phase: Add tanaka ethanol extract, angelica ethanol extract and fangfeng ethanol extract to lotus seed shell ethanol extract, stir evenly to obtain a mixed extract; add olive oil, glycerin and butylene glycol, stir and mix evenly; (b) Preparation of emulsion aqueous phase: stirring and mixing purified water, soybean lecithin, carbomer and glycerin; (c) Preparation of emulsion: slowly add the emulsion oil phase to the emulsion water phase under stirring to form an emulsion; after emulsification, cool the emulsion and adjust the pH value to obtain an emulsion.

[0017] Preferably, in the oil phase of the emulsion, the volume ratio of lotus seed shell ethanol extract, tanaka ethanol extract, angelica ethanol extract and saposhnikovia ethanol extract is 10-8:5-4:2-1:1; the added amounts of olive oil, glycerol and butylene glycol are 2 times, 1 times and 0.5 times the mass of the mixed extract, respectively; in the water phase of the emulsion, purified water, soybean lecithin, carbomer and glycerol are 10 times, 40%, 10% and 1 times the mass of the mixed extract, respectively.

[0018] Preferably, in steps (a) to (c), the stirring temperature is 70° C.; the emulsion is cooled to 40-45° C., and triethanolamine is added to adjust the pH value of the emulsion to 5.5-6.0.

[0019] The present invention uses lotus seed shell as the main raw material to develop a UV protection combination product combined with natural lotus seed shell extract. The product not only contains an external emulsion, but also innovatively introduces the concept of oral sunscreen beverage. This internal and external combined protection method not only improves the skin's external barrier to ultraviolet rays, but also enhances the skin's internal antioxidant capacity through oral administration, achieving comprehensive protection of the skin.

[0020] The beneficial effects of the present invention are: (1) The extraction method provided by the present invention has high extraction efficiency, and through fermentation treatment, the free radical scavenging effect of the extract can be improved, and the extraction efficiency is high; and the method can reduce the damage to the compounds in the extraction process, and the content of the extracted active ingredients is high, which is conducive to the effect of lotus seeds; (2) The method provided by the present invention is simple and easy to operate, and can be widely applied in industry. The prepared supernatant and ethanol extract have significant anti-ultraviolet effects and can scavenge a variety of free radicals. (3) The UV protection combination product prepared by the present invention not only improves the skin's external barrier to UV rays through a combination of internal and external protection, but also enhances the skin's intrinsic antioxidant capacity through oral administration, thereby achieving comprehensive protection for the skin. DETAILED DESCRIPTION

[0021] The technical solution of the present invention is further explained and illustrated by means of specific embodiments below.

[0022] The tanaka ethanol extract, angelica ethanol extract and saposhnikovia ethanol extract used in the present invention are prepared by the following method: adding anhydrous ethanol to tanaka, angelica or saposhnikovia at a mass ratio of 1:10, ultrasonically extracting for 30 minutes at an ultrasonic frequency of 40kHz, a power of 100W and room temperature, and then concentrating to 1 / 7 of the original volume to obtain the product.

[0023] Example 1 (1) Raw material processing: select fresh lotus seed shells, wash and remove impurities, dry the lotus seed shells, and then use a grinder to grind them into a particle size of 100 mesh to obtain lotus seed shell powder; (2) Fermentation treatment: Mix lotus seed shell powder and water in a ratio of 1:12, then inoculate Aspergillus oryzae and Bacillus pumilus in a ratio of 2:1, and the total inoculation amount is 3% of the total amount of lotus seed shell. After stirring evenly, add betaine accounting for 0.15% of the total amount of lotus seed shell powder, stir evenly, and ferment at a fermentation temperature of 28-30°C and a fermentation time of 72 hours to obtain a fermentation liquid; (3) Post-fermentation treatment: Sterilize the fermentation broth at 121°C for 20 min. After cooling, homogenize it in a homogenizer at 10,000-12,000 rpm for 1 min. Repeat 5 times.

[0024] (4) Centrifugation: After homogenization, centrifuge the sample at 6000 rpm for 30 min, and collect the supernatant and precipitate respectively; (5) Precipitation extraction: The precipitate obtained in step (4) was dried, and anhydrous ethanol 10 times its mass was added for ultrasonic-assisted extraction. The ultrasonic frequency was 40 kHz, the power was 100 W, the extraction time was 30 min, and the extraction temperature was room temperature. After the extraction was completed, the precipitate was concentrated to 1 / 8 of the original volume to obtain the lotus seed shell ethanol extract.

[0025] Comparative Example 1 (1) Raw material processing: select fresh lotus seed shells, wash and remove impurities, dry the lotus seed shells, and then use a grinder to grind them into a particle size of 100 mesh to obtain lotus seed shell powder; (2) Fermentation treatment: Mix lotus seed shell powder and water in a ratio of 1:12, then inoculate Aspergillus oryzae and Bacillus pumilus in a ratio of 2:1, and the total inoculation amount is 3% of the total amount of lotus seed shells. After stirring evenly, fermentation is carried out: the fermentation temperature is 28-30°C, the fermentation time is 72h, and the fermentation liquid is obtained; The other steps are the same as in Example 1.

[0026] Comparative Example 2 (1) Raw material processing: select fresh lotus seed shells, wash and remove impurities, dry the lotus seed shells, and then use a grinder to grind them into a particle size of 100 mesh to obtain lotus seed shell powder; (2) Precipitation extraction: Add 10 times the mass of anhydrous ethanol to the lotus seed shell powder and perform ultrasonic-assisted extraction. The ultrasonic frequency is 40 kHz, the power is 100 W, the extraction time is 30 min, and the extraction temperature is room temperature. After the extraction is completed, the lotus seed shell is concentrated to 1 / 8 of the original volume to obtain the lotus seed shell ethanol extract.

[0027] Example 2 Preparation of emulsion (1) Preparation of emulsion oil phase: Add tanaka ethanol extract, angelica dahurica ethanol extract and saposhnikovia divaricata ethanol extract to the lotus seed shell ethanol extract in a ratio of 10:5:2:1, add 2 times the weight of the mixed extract of olive oil, 1 times the weight of glycerin, and 50% of the weight of butylene glycol, and stir and mix evenly at 70°C.

[0028] (2) Preparation of emulsion aqueous phase: 10 times the mass of pure water of the mixed extract, 40% the mass of soybean lecithin of the mixed extract, 10% of carbomer of the mixed extract and 1 times the mass of glycerol were stirred and mixed at 70°C.

[0029] (3) Preparation of emulsion: The heated oil phase in step 7 is slowly added to the aqueous phase in step 8 under stirring at 800-1000 rpm to form an emulsion. After emulsification, the emulsion is cooled to 45° C. Triethanolamine is added to adjust the pH value of the emulsion to 6.0, and an emulsion containing the ethanol extract of lotus seed shell is obtained.

[0030] Example 3 Preparation of Beverages The supernatant is added to purified water at a ratio of 2:1, and then 0.5% ascorbic acid, 0.01% sodium selenate, 0.05% zinc sulfate, and 1% stevioside are added to the total volume of the mixture. After evenly mixing, the mixture is filtered, homogenized, degassed, sterilized, and other steps to form a beverage containing the supernatant.

[0031] Effect Example 1 (1) Preparation of DPPH reagent Weigh DPPH powder, place it in a 250mL volumetric flask, dissolve it in 95% ethanol to make up to 250mL, and obtain a DPPH reagent with a concentration of 0.06mmol / L.

[0032] (2) Preparation of test samples The lotus seed shell ethanol extract prepared in Example 1 and Comparative Examples 1-2 was added with 95% ethanol to prepare a 0.01 g / mL test solution.

[0033] The same method was used to prepare a 1 mg / mL vitamin E control solution.

[0034] (3) Determination of antioxidant capacity a. Add 4.0mL of DPPH solution and 1.0mL of 95% ethanol to a test tube, mix and shake well, and react for 30 minutes in the dark. After stabilization, measure the absorbance at 517nm with 95% ethanol as a reference, and record it as A0.

[0035] b. Add 4.0mL DPPH solution and 1.0mL test solution into the test tube, mix and shake well, and react for 30 minutes in the dark. After stabilization, measure the absorbance at 517nm with 95% ethanol as reference, and record it as A. r .

[0036] c. Add 4.0mL of 95% ethanol solution and 1.0mL of the sample solution to be tested into the test tube, mix and shake well, and react for 30 minutes in the dark. After stabilization, measure the absorbance at 517nm with 95% ethanol as the reference, and record it as A. s .

[0037] d. The scavenging rate of the sample on DPPH free radicals is calculated according to the following formula: The test results are shown in Table 1 below.

[0038] Table 1 Effect Example 2 Test method: The lotus seed shell ethanol extract and supernatant prepared in Example 1 and Comparative Examples 1-2 were prepared into 600 mg / mL and 800 mg / mL test concentration groups using aquaculture water. One parallel group was set for each concentration, and another blank control group of aquaculture water was set.

[0039] During the experiment, healthy and active adult zebrafish were selected and put into the relevant groups. Ten zebrafish were randomly put into each beaker. No food was fed during the whole experiment to avoid the influence of feed. The laboratory temperature was controlled at (24±1)℃, and the medicine was changed once every 24 hours. After the experiment started, continuous observation was carried out. If the fish died, they were fished out in time and the experiment was recorded. The observation was ended at 96 hours, and data statistics and analysis were performed.

[0040] It was found through experiments that in the embodiments and comparative examples, the lotus seed shell ethanol extract and the supernatant prepared by the preparation method provided by the present invention had no obvious effect on zebrafish, indicating that the extract provided by the present invention had no acute toxic reaction to zebrafish and had good safety.

[0041] Effect Example 3 Establishment of NIH3T3 photoaging model: NIH3T3 cells were cultured in DMEM medium containing 1% double antibody and 10% fetal bovine serum. When the cells grew to the logarithmic growth phase, 2×10 4 The cells were inoculated into culture dishes and six-well plates, respectively, and cultured at 37°C and 5% CO2 for 24 h before being irradiated with different doses of UVA. Aluminum foil was used to wrap the UVA irradiator and the cells (irradiation dose = irradiation intensity × irradiation time; irradiation intensity was 0.25 J / cm 2 ) The control group was not irradiated. After irradiation, the cells were washed once with physiological saline and cultured in DMEM containing 1% fetal bovine serum. After 24 hours, 10% CCK-8 reagent was added to measure the cell survival rate in the dark. The UVA irradiation dose with a cell survival rate of about 50% of that of cells without UVA irradiation was selected for subsequent experiments.

[0042] The CCK-8 method was used to detect the effect of lotus seed shell ethanol extract on the proliferation rate of NIH3T3 cells. During the continuous culture of cells irradiated by UVA, 50 μg / mL of extracts (Example 1 and Comparative Examples 1 and 2) were added respectively, and the results were analyzed after 24 hours of action. The specific results are shown in Table 2 below.

[0043] Table 2

Claims

1. A method for preparing a lotus seed shell extract having an anti-ultraviolet function, characterized in that: The following steps are involved: (1) Fermentation treatment: Mix lotus seed shell powder with water, inoculate fermentation agent, add a small amount of betaine, stir evenly and ferment to obtain fermentation liquid; (2) Post-fermentation treatment: sterilize and cool the fermentation broth, then homogenize and centrifuge it, collect the supernatant and precipitate respectively; dry the precipitate for later use; (3) Precipitation extraction: Add 8-10 times the mass of anhydrous ethanol to the dried precipitate and perform ultrasonic extraction. After the extraction, concentrate the precipitate to obtain the lotus seed shell ethanol extract.

2. The preparation method according to claim 1, characterized in that: In step (1), the mass ratio of the lotus seed shell powder to water is 1:10-1:15; the fermentation agent is composed of Aspergillus oryzae and Bacillus pumilus in a mass ratio of 2:1; the inoculation amount of the fermentation agent accounts for 2%-3% of the total amount of the lotus seed shell powder; and the added amount of betaine accounts for 0.1%-0.2% of the total amount of the lotus seed shell powder.

3. The preparation method according to claim 1 or 2, characterized in that: In step (1), the fermentation is carried out at a temperature of 28-30° C. and a fermentation time of 60-72 hours.

4. The preparation method according to claim 1, characterized in that: In step (2), the speed of the homogenizer is 10000-12000 rpm for 1 min, which is repeated 5-6 times; the speed of the centrifuge is 6000-8000 rpm, and the centrifugation time is 30 min.

5. The preparation method according to claim 1 or 4, characterized in that: In step (3), the mass ratio of the precipitate to anhydrous ethanol is 1:8-10; the conditions of the ultrasonic extraction are: ultrasonic frequency 40 kHz, power 100 W, extraction time 30 min, extraction temperature is room temperature; and the extract is concentrated to 1 / 7-1 / 8 of the original volume.

6. Use of the lotus seed shell extract prepared by the preparation method according to any one of claims 1 to 5 in the preparation of anti-ultraviolet products, characterized in that: The anti-ultraviolet product comprises a beverage containing a supernatant and an emulsion containing an ethanol extract of lotus seed shell.

7. The use according to claim 6, characterized in that: The beverage containing the supernatant is prepared by the following method: adding the supernatant to purified water at a ratio of 2:1 to obtain a mixed solution; then adding 0.5% ascorbic acid, 0.01% sodium selenate, 0.05% zinc sulfate, and 1% stevioside, accounting for the total weight of the mixed solution, mixing evenly, filtering, homogenizing, degassing, and sterilizing to obtain the beverage.

8. The use according to claim 6, characterized in that: The emulsion containing the lotus seed shell ethanol extract is prepared by the following method: (a) Preparation of the emulsion oil phase: Add the ethanol extract of tanaka, the ethanol extract of angelica and the ethanol extract of fangfeng to the ethanol extract of lotus seed shell, stir evenly to obtain a mixed extract; add olive oil, glycerin and butylene glycol, stir and mix evenly; (b) Preparation of the aqueous phase of the emulsion: 10 times the mass of pure water of the mixed extract, 40% the mass of soybean lecithin of the mixed extract, 10% of carbomer of the mixed extract and 1 times the mass of glycerol are stirred and mixed uniformly; (c) Preparation of emulsion: Slowly add the emulsion oil phase into the emulsion water phase under stirring, stir and emulsify, cool the emulsion and adjust the pH value to obtain the emulsion.

9. The use according to claim 8, characterized in that: In the oil phase of the emulsion, the volume ratio of lotus seed shell ethanol extract, tanaka ethanol extract, angelica ethanol extract and saposhnikovia ethanol extract is 10-8:5-4:2-1:1; the added amounts of olive oil, glycerol and butylene glycol are 2 times, 1 time and 0.5 times the mass of the mixed extract respectively; in the water phase of the emulsion, the purified water, soybean lecithin, carbomer and glycerol are 10 times, 40%, 10% and 1 time the mass of the mixed extract respectively.

10. Use according to claims 8 and 9, characterized in that In steps (a) to (c), the stirring temperature is 70° C.; the emulsion is cooled to 40-45° C., and triethanolamine is added to adjust the pH value of the emulsion to 5.5-6.0.

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