Preparation of hericium erinaceus natural component with whitening effect and prepared hericium erinaceus natural component

Abstract

The invention discloses preparation of a hericium erinaceus natural component with a whitening effect and the prepared hericium erinaceus natural component. The preparation method comprises the following steps: (1) crushing byproducts generated in the processing process of hericium erinaceus sporocarp with a crusher, sieving to obtain powder, soaking a proper amount of powder with an ethanol aqueous solution, carrying out ultrasonic assistance during the period, centrifugally collecting filtrate, carrying out rotary evaporation on the filtrate to remove a solvent to obtain concentrate, and mixing the concentrate with pure water to obtain a mixed solution; a concentrated solution is obtained; (2) filling activated H103 macroporous adsorption resin into a chromatographic column, filling the concentrated solution into the H103 macroporous adsorption resin until the adsorption is saturated, and initially using purified water to flow through the column; and eluting with a low-grade ethanol aqueous solution, collecting a low-polarity eluent, carrying out rotary evaporation, and freeze-drying to obtain a low-polarity eluate. The process is simple and efficient in steps, low in cost and safe, and the obtained product has a good whitening effect.

Description

(I) Technical field

[0001] The invention relates to the technical field of cosmetic formulas, and in particular to a method for preparing a natural whitening active ingredient extracted from Hericium erinaceus and the prepared Hericium erinaceus natural ingredient. (II) Technical Background

[0002] Hericium erinaceus, also known as Hericium erinaceus, Lion's mane mushroom, Hedgehog mushroom, etc., is rich in nutrients and has the advantages of high protein and low fat. It mainly grows in Northeast China, North China and East China. As a precious edible mushroom with both medicinal and edible properties, Hericium erinaceus has a long history in China and is widely used in cooking and traditional Chinese medicine. However, it is currently mainly sold in the form of dried products on the market, and there is a lack of deep-processed products, which is not conducive to its development in the modern edible fungus industry. In order to promote the development of the edible fungus industry towards modernization, functionalization, convenience and personalization, it is necessary to strengthen the extraction of Hericium erinaceus functional substances and develop deep-processed products to improve the utilization efficiency and value of Hericium erinaceus.

[0003] In recent years, with the increase in edible fungi enterprises, a large number of by-products are produced in the processing of Hericium erinaceus fruiting bodies. These by-products are rich in nutrients such as terpenoids and flavonoids, but are often discarded, leading to increasingly serious environmental pollution and waste of resources. With the rapid development of my country's agriculture and food industry, higher requirements are placed on environmental protection and resource utilization. Through deep processing of Hericium erinaceus by-products, not only can environmental pollution be reduced, but the active ingredients in the Hericium erinaceus fruiting body by-products can be more fully utilized, achieving efficient resource utilization and promoting sustainable development of the industry.

[0004] Terpenoids are a class of compounds composed of isoprene (C5H8) and its derivatives, which are widely found in plants and fungi. Studies have shown that terpenoids have a variety of pharmacological activities such as antioxidant and anti-aging. Terpenoids in Hericium erinaceus have been shown to have antibacterial and enzyme inhibitory properties. Although there are many active ingredients that can be used in whitening products, there are relatively few with practical application value, such as hydroquinone and β-arbutin. These inhibitors are not only expensive, but also have certain toxic side effects.

[0005] Studies have shown that due to their significant whitening effects and pharmacological properties, terpenoids have been widely used in a variety of whitening product formulas. For example, patent CN202310730990.3 discloses a whitening and moisturizing plant essential oil composition containing various plant essential oils such as star anise essential oil and a preparation method thereof. The moisturizing essential oil is composed of star anise essential oil, galangal essential oil, ylang ylang essential oil, sandalwood essential oil, borage essential oil, pyridine and stearyl chloride. These plant essential oil ingredients improve the biocompatibility and moisturizing efficacy of the product, but their production cost is relatively high, the processing cost is high, and some raw materials are irritating, and long-term use may have adverse effects on consumers' skin.

[0006] Patent CN 202210081172.0 discloses a gel-type whitening mask based on Ophiopogon japonicus and its preparation method, which uses Ophiopogon japonicus high isoflavone extract, sodium hyaluronate, allantoin, etc. as raw materials. This gel can effectively inhibit tyrosinase activity and reduce dark spots caused by melasma and acne, but its preparation process is complicated, and a large number of active ingredients may be secreted, decomposed or unstable after encapsulation. (III) Summary of the invention

[0007] The present invention aims to overcome the defects in the prior art and proposes a method for preparing a natural component of Hericium erinaceus with whitening effect and the prepared natural component of Hericium erinaceus with whitening effect. The method has the advantages of simple process steps, stable and controllable process, green and environmental protection, etc., improves the safety and stability of the product, and provides practical support for the resource utilization of Hericium erinaceus, further enhancing its economic value.

[0008] To achieve this object, the present invention adopts the following technical solutions:

[0009] In a first aspect, the present invention provides a method for preparing a natural component of Hericium erinaceus having whitening effect, comprising the following steps:

[0010] (1) crushing the byproducts generated during the processing of Hericium erinaceus fruiting bodies with a pulverizer, sieving to obtain powder, taking an appropriate amount of the powder and soaking it in a 60% by volume ethanol aqueous solution, during which ultrasonic assistance is provided, each ultrasonic temperature is 50 to 55° C., and the ultrasonic time is 30 to 50 min; collecting the filtrate by centrifugation, taking the filtrate and performing rotary evaporation to remove the solvent to obtain a concentrate, taking the concentrate and mixing it with pure water to obtain a concentrated solution;

[0011] (2) Loading the activated H103 macroporous adsorption resin into a chromatography column, and filling the H103 macroporous adsorption resin with the concentrated solution obtained in step (1) until adsorption is saturated, and initially using pure water to flow through the column to remove non-specifically adsorbed impurities; then using a 20% by volume low-polarity ethanol aqueous solution for elution, collecting an equal volume of low-polarity eluate based on the volume of the elution reagent, and performing rotary evaporation until it becomes a paste, and freeze-drying the paste to obtain a low-polarity eluate, which is a natural ingredient of Hericium erinaceus with whitening effect.

[0012] Preferably, in step (1), the centrifugal speed is 8000-12000 r / min, and the centrifugal time is 10-20 min.

[0013] Preferably, in step (1), the solid-liquid ratio of the powder to 60% by volume ethanol aqueous solution is 1 g: (10-20) mL.

[0014] Preferably, in step (1), the concentrate is mixed with pure water in a mass ratio of 1:(3-5).

[0015] Preferably, in step (2), the activation method of the H103 macroporous adsorption resin is as follows: soak the H103 macroporous adsorption resin in 2 volumes of 95% ethanol solution for 18 to 24 hours, then wash with distilled water until there is no alcohol smell; soak it in 2 volumes of 5% to 10% sodium hydroxide aqueous solution by mass for 1 to 2 hours, then adjust to neutral with distilled water; finally treat it with 5% to 10% hydrochloric acid solution by mass for 1 to 2 hours, and wash it to neutral.

[0016] Preferably, in step (2), 3 to 5 BV of pure water is initially passed through the column to remove non-specifically adsorbed impurities.

[0017] Preferably, in step (2), the volume of the 20% by volume low-grade ethanol aqueous solution is 2 to 3 times that of the concentrated solution, more preferably 2 times.

[0018] Preferably, the rotary evaporation temperature in step (2) is 55-65°C, more preferably 55-60°C.

[0019] Preferably, the preparation method further comprises the following step (3):

[0020] (3) After all the low-polarity eluent in step (2) is collected, it is immediately replaced with a 40% by volume medium-polarity ethanol aqueous solution to continue elution. An equal volume of the medium-polarity eluent is collected based on the volume of the elution reagent, and rotary evaporated until it becomes a paste. The paste is freeze-dried to obtain a medium-polarity eluate, which is a natural ingredient of Hericium erinaceus with whitening effect.

[0021] As further preferred, in step (3), the volume usage of the 40% by volume aqueous solution of neutral polar ethanol is 2 to 3 times that of the concentrated solution, and more preferably 2 times.

[0022] Preferably, the rotary evaporation temperature in step (3) is 55-65°C, more preferably 55-60°C.

[0023] As a further preference, the preparation method further comprises step (4):

[0024] (4) After all the intermediate eluent is collected, it is immediately replaced with a 60% high-polarity ethanol aqueous solution to continue elution. An equal volume of high-polarity eluent is collected based on the volume of the elution reagent, and rotary evaporated until it becomes a paste. The paste is freeze-dried to obtain a high-polarity eluate, which is a natural ingredient of Hericium erinaceus with whitening effect.

[0025] Preferably, in step (4), the volume of the 60% by volume high-polarity ethanol aqueous solution is 2 to 3 times that of the concentrated solution, more preferably 2 times.

[0026] Preferably, the rotary evaporation temperature in step (4) is 55-65°C, more preferably 55-60°C.

[0027] In a second aspect, the present invention provides a natural ingredient of Hericium erinaceus with whitening effect prepared according to the preparation method described in the first aspect.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] (1) The process steps are simple and efficient: The preparation method of the present invention extracts valuable components from the by-products of fresh Hericium erinaceus and purifies them by eluting with ethanol solutions of different polarities, showing the characteristics of simple process and high efficiency.

[0030] Specifically, compared with conventional technical processes, the present invention adopts gradient ethanol elution technology, which can efficiently obtain the bioactive components in Hericium erinaceus. And the conventional technical process requires a large number of instruments and equipment, such as semi-preparative liquid chromatography and other instruments, the operation is relatively complicated, and the time required for industrial production is usually 1 week or more. The process flow of the present invention is relatively simple, and the time required for the method of the present invention to obtain a medium-polarity eluate under laboratory conditions is only 24 hours, and the efficiency is improved by 85.71%. See the process flow comparison diagram Figure 1 .

[0031] (2) Multifunctionality: The polar eluate obtained by the present invention not only has a whitening effect that is superior to the commercially available whitening active substance β-arbutin, but also has significantly better antioxidant properties than β-arbutin, and can provide all-round health support for the fields of skin care products and health care products. Experimental data show that the medium-polarity eluate has strong antioxidant and tyrosinase inhibition activity and has the best effect in inhibiting tyrosinase activity, and can be used as a potential main active ingredient of whitening cosmetics.

[0032] (3) Low cost and safety: The Hericium erinaceus extraction process of the present invention exhibits significant advantages in terms of cost and safety, forming a complementary system, which is an important factor in improving market competitiveness.

[0033] Specifically, compared with the conventional process, this method optimizes the extraction steps, mainly using ethanol and aqueous solution, and the cost of extracting each gram of active ingredient is controlled below RMB 5, a reduction of 50%. It only takes 24 hours to obtain a medium-polarity eluate, which greatly improves work efficiency and reduces worker costs. In addition, the raw material price of Hericium erinaceus is low, and the commercial price of Hericium erinaceus is between RMB 20 and 30. In terms of safety, this method is based on natural materials and does not add any high-risk chemical reagents, ensuring product safety and meeting green cosmetics standards. This comprehensive advantage will promote the market circulation of cost-effective extracts and bring good economic benefits. (IV) Description of the drawings

[0034] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0035] Figure 1 The invention is an implementation method of the separation and purification process of Hericium erinaceus fruiting bodies by eluting with ethanol of different volume percentages.

[0036] Figure 2 The scavenging ability of the products of Examples 1-4 and β-arbutin on DPPH free radicals.

[0037] Figure 3 The scavenging ability of the products of Examples 1-4 and β-arbutin on ABTS free radicals.

[0038] Figure 4 The inhibitory effects of the products of Examples 1-4 and β-arbutin on the activity of tyrosine monophenolase are shown.

[0039] Figure 5 The inhibitory effects of the products of Examples 1-4 and β-arbutin on the activity of tyrosine diphenolase are shown. (V) Specific implementation methods

[0040] The present invention is further explained below in conjunction with specific examples. The present invention is implemented according to the technical solution of the invention, and detailed implementation methods and operating steps are given, but the protection scope of the present invention is not limited to the following examples. The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions.

[0041] Example 1

[0042] See also Figure 1 , a method for preparing a natural whitening active ingredient extracted from Hericium erinaceus, the specific steps are as follows:

[0043] Step 1: preparing ethanol aqueous solution: prepare ethanol aqueous solutions of different polarities (volume percentage 20%, 40%, 60%, 95%) for subsequent steps.

[0044] Step 2. Grinding and sieving Hericium erinaceus: Take the remaining byproducts produced during the processing of Hericium erinaceus fruiting bodies and grind them with a grinder. Pass the crushed Hericium erinaceus byproducts through a 100-mesh sieve and weigh an appropriate amount of powder.

[0045] Step 3. Ultrasonic-assisted ethanol extraction: The weighed Hericium erinaceus powder is soaked in an ethanol-water solution with a volume percentage of 60%, and the solid-liquid ratio is 1 g: 15 mL. Ultrasonic assistance is performed during the soaking process, and each ultrasonic temperature is 55° C. and the ultrasonic time is 30 minutes.

[0046] Step 4: Centrifugal extraction: After the ultrasonic extraction, the mixed solution was centrifuged at a rate of 12000 r / min for 10 min. The filtrate was collected and combined to obtain 600 mL of the filtrate, and the clear upper liquid was obtained and refrigerated at 4°C for later use.

[0047] Step 5: Rotary evaporation: Take the supernatant of step 4 and operate it in a rotary evaporator at 55°C until the ethanol is completely evaporated to obtain the Hericium erinaceus concentrate. Mix the concentrate with pure water in a ratio of 1:3 to obtain 900 mL of concentrated solution, and refrigerate at 4°C for later use.

[0048] Step 6: Hericium erinaceus concentrate: Take part of the crude extract of step 5 and freeze-dry (the cold trap temperature is -80°C, the freezing chamber temperature is -45°C, the vacuum degree is 6.8 Pa, and the freeze-drying time is 2 days) to obtain Hericium erinaceus concentrate.

[0049] Example 2

[0050] Step 1-5: Same as Example 1.

[0051] Step 6: Activation treatment of H103 macroporous resin: Weigh 350g of H103 macroporous resin, soak it in 95% ethanol solution for 24h, then wash it with distilled water until there is no alcohol smell; then soak it in 5% sodium hydroxide solution for 12h, then adjust it to neutral with distilled water; finally treat it with 5% hydrochloric acid for 12h and wash it to neutral.

[0052] Step 7: Resin adsorption and elution: The activated resin was loaded into a chromatography column, and about 500 mL of the Hericium erinaceus concentrate obtained in step 5 was filled into the H103 macroporous resin at a flow rate of 3 BV / h until adsorption saturation was reached.

[0053] Initial elution: Initially use 5 BV of pure water to flow through the column to remove non-specifically adsorbed impurities.

[0054] The first stage of low-grade elution: gradient elution is performed, and the first elution is performed using a low-grade ethanol solution with a volume of 1 L and a volume percentage of 20%.

[0055] Collect the eluent: Collect an equal volume of low polarity eluent based on the eluent volume.

[0056] Step 8: Rotary evaporation: Take the low-polarity eluent obtained in step 7, and evaporate it by rotary evaporation (the temperature of the constant temperature water tank is 60°C, the time is 25 min, the speed is 80 rpm, and the vacuum degree is -0.1 MPa) until it becomes a paste, and freeze-dry the paste (the temperature of the cold trap is -80°C, the temperature of the freezing chamber is -45°C, the vacuum degree is 6.8 Pa, and the freeze-drying time is 2 days) to obtain a low-polarity eluate.

[0057] Example 3

[0058] Step 1-7: Same as Example 2.

[0059] Step 8: Secondary intermediate elution: In step 7, after all the low-polarity eluent is collected, it is immediately replaced with a 1 L, 40% by volume intermediate ethanol solution to continue elution.

[0060] Collect the eluent: Collect equal volumes of the neutral polarity eluent based on the eluent volume.

[0061] Step 9: Rotary evaporation: Take the intermediate polar eluent of step 8, and evaporate it by rotary evaporation (the temperature of the constant temperature water bath is 60°C, the time is 25 min, the speed is 80 rpm, and the vacuum degree is -0.1 MPa) until it becomes a paste, and freeze-dry the paste (the temperature of the cold trap is -80°C, the temperature of the freezing chamber is -45°C, the vacuum degree is 6.8 Pa, and the freeze-drying time is 2 days) to obtain the intermediate polar eluate.

[0062] Example 4

[0063] Step 1-8: Same as Example 3.

[0064] Step 9: Three-stage high-polarity elution: In step 8, after all the intermediate eluent is collected, it is immediately replaced with a 1L, 60% by volume high-polarity ethanol solution to continue elution.

[0065] Collect the eluent: Collect an equal volume of the high polarity eluent according to the eluent volume.

[0066] Step 10: Rotary evaporation: Take the high-polarity eluent of step 9, and evaporate it by rotary evaporation (the temperature of the constant temperature water bath is 60°C, the time is 25 min, the speed is 80 rpm, and the vacuum degree is -0.1 MPa) until it becomes a paste, and freeze-dry the paste (the temperature of the cold trap is -80°C, the temperature of the freezing chamber is -45°C, the vacuum degree is 6.8 Pa, and the freeze-drying time is 2 days) to obtain a high-polarity eluate.

[0067] The eluate prepared by the above steps has good antioxidant activity and tyrosinase inhibition activity, and can be suitable for the preparation of whitening products and other related applications.

[0068] Embodiment 5:

[0069] (1) The products of Examples 1-4 were dissolved in PBS:DMSO (9:1 (v / v)) for subsequent application research. The absorbance at 547 nm was determined according to the vanillin-perchloric acid method, and a standard curve was prepared using oleanolic acid solutions of different mass concentrations to determine the content of terpenoid compounds in the products of Examples 1-4. The sample solutions to be tested were obtained by gradient dilution according to the concentration of terpenoid compounds in the products of Examples 1-4.

[0070] (2) Determination of the DPPH free radical scavenging ability of the products of Examples 1-4: First, prepare 0.05 mg / mL DPPH; mix 100 μL of the sample solution of step (1) with different gradient concentrations and 100 μL of DPPH ethanol solution in a 96-well plate, and then measure the absorbance at a wavelength of 517 nm using an ELISA reader.

[0071] (3) Determination of the ABTS free radical scavenging ability of the products of Examples 1-4: First, prepare 7.4 mmol / L ABTS diammonium salt stock solution and 2.6 mmol / L potassium persulfate stock solution, mix the two solutions in equal volumes, and place them at room temperature in a dark environment for 12 hours, dilute them with pH 7.4 PBS:DMSO (9:1 (v / v)) at a ratio of 1:20 (v:v) to obtain ABTS working solution, the detection value of which at 734 nm on an ELISA reader is about 0.7; 100 μL of the sample solution to be tested in step (1) with different gradient concentrations and 100 μL ABTS working solution are mixed in a 96-well plate, and then the absorbance is measured at a wavelength of 734 nm on an ELISA reader.

[0072] (4) Determine the ability of the products of Examples 1-4 to inhibit the in vitro activity of tyrosine monophenolase: First, prepare a 0.5 g / L L-tyrosine solution. Accurately weigh 0.5 g L-tyrosine, add the prepared PBS buffer (50 mmol / L, pH 6.8) to dissolve, and dilute to 1000 mL; secondly, prepare a 100 U / mL tyrosinase solution. Accurately weigh 1000 U tyrosinase, add the prepared PBS buffer (50 mmol / L, pH 6.8) to dissolve, and dilute to 1000 mL. Store the prepared L-tyrosine solution at 4°C away from light for future use. 80 μL of the sample solution of step (1) with different gradient concentrations was added to a 96-well ELISA plate, and then 100 μL of L-tyrosine solution pre-warmed at 37°C was added, and the mixture was shaken and mixed. After being placed in a 37°C constant temperature box for 10 min, 50 μL of tyrosinase solution was added, and the mixture was mixed. The mixture was allowed to react fully at 37°C for 10 min, and the absorbance was measured at 475 nm.

[0073] (5) Determination of the in vitro tyrosine diphenolase inhibitory ability of the products of Examples 1-4: First, prepare a 1.0 g / L L-dopa (L-DOPA) solution. Accurately weigh 1.0 g L-DOPA, add the prepared PBS buffer (50 mmol / L, pH 6.8) to dissolve, and dilute to 1000 mL; secondly, prepare a 100 U / mL tyrosinase solution. Accurately weigh 1000 U tyrosinase, add the prepared PBS buffer (50 mmol / L, pH 6.8) to dissolve, and dilute to 1000 mL. Store the prepared L-DOPA solution at 4°C in the dark for future use. 80 μL of the sample solution of step (1) with different gradient concentrations was added to a 96-well ELISA plate, and then 100 μL of L-DOPA solution pre-warmed at 37°C was added, and the mixture was shaken and mixed. After being placed in a 37°C constant temperature box for 10 min, 50 μL of tyrosinase solution was added, and the mixture was mixed. The mixture was allowed to react fully at 37°C for 10 min, and the absorbance was measured at 475 nm.

[0074] Comparative Example 1:

[0075] Take β-arbutin as positive control: accurately weigh 200 mg of β-arbutin, and dilute to volume with 100 mL of PBS:DMSO (9:1, v / v) to obtain 2 mg / mL β-arbutin, and then gradient dilute to a specific concentration. Perform performance test according to Example 5, except that the experimental sample is replaced by the β-arbutin sample.

[0076] The test results are as follows:

[0077] In vitro antioxidant activity test: The antioxidant activity of the gradient concentration samples prepared in Example 5 and Comparative Example 1 was studied, and the results are as follows: Figure 2 and Figure 3 As shown. The results of the DPPH method and the ABTS method are both expressed in terms of antioxidant rate (%). Combining the results of the above two methods, it can be seen that the antioxidant activities of different examples determined by the DPPH method are in the following order from large to small: medium polarity eluate [Example 3] > low polarity eluate [Example 2] > high polarity eluate [Example 4] > Hericium erinaceus concentrate [Example 1] > β-arbutin [Example 5]. Among them, the medium polarity eluate [Example 3] has the largest antioxidant rate among these eluates, and the high polarity eluate [Example 4] has the smallest antioxidant rate, and all eluates have better DPPH free radical scavenging effects than β-arbutin. Specifically, the IC value of the medium polarity eluate for DPPH free radical scavenging rate is 50 The value was 64.77 μg / mL, which was significantly better than β-arbutin (IC 50 The value is 156.50 μg / mL). The antioxidant activities of different examples determined by the ABTS method are in the following order from small to large: β-arbutin [Example 5] > medium polarity eluate [Example 3] > low polarity eluate [Example 2] > high polarity eluate [Example 4] > Hericium erinaceus concentrate [Example 1]. Among them, the medium polarity eluate [Example 3] has the largest antioxidant rate among these eluates, while the high polarity eluate [Example 4] has the smallest antioxidant rate. Moreover, the ABTS free radical scavenging effect of all eluates is better than that of the alcohol extraction rotary evaporation liquid. Specifically, the IC value of the medium polarity eluate on the ABTS free radical scavenging rate is 50 The value was 0.66 μg / mL, which was significantly better than β-arbutin (IC 50 The value is 0.86 μg / mL). Combining the results measured by the above two methods, the anti-oxidation effect of the medium polarity eluate [Example 3] is the best.

[0078] In vitro tyrosinase activity inhibition: The gradient concentration samples prepared in Example 5 and Comparative Example 1 were subjected to in vitro tyrosinase activity inhibition studies. The results are as follows: Figure 4 and Figure 5 As shown. The results of in vitro inhibition of tyrosine monophenolase activity and tyrosine diphenolase activity are both expressed in terms of enzyme activity inhibition rate (%). Combining the results measured by the above two methods, it can be seen that for the results of in vitro inhibition of tyrosine monophenolase activity, the order of the inhibition effects of tyrosine monophenolase activity of different embodiments measured is from large to small: β-arbutin [Example 5] > medium polarity eluate [Example 3] > Hericium erinaceus concentrate [Example 1] > high polarity eluate [Example 4] > low polarity eluate [Example 2], among which the medium polarity eluate [Example 3] has the largest inhibition rate for tyrosine monophenolase among these embodiments, and the low polarity eluate [Example 2] has the smallest inhibition rate. Specifically, the IC value of the medium polarity eluate on the inhibition rate of tyrosine monophenolase activity 50The value was 0.87 μg / mL, which was significantly better than β-arbutin (IC 50 The value is 0.71 μg / mL). The results of the inhibition of tyrosine diphenolase in vitro activity showed that the inhibition effects of different examples on tyrosine diphenolase activity were in the following order from large to small: medium polarity eluate [Example 3] > high polarity eluate [Example 4] > Hericium erinaceus concentrate [Example 1] > low polarity eluate [Example 2] > β-arbutin [Example 5]. Among them, the medium polarity eluate [Example 3] had the largest inhibition rate on tyrosine diphenolase among these extracts, while the low polarity eluate [Example 2] had the smallest inhibition rate, and β-arbutin did not have an inhibitory effect on the activity of tyrosine diphenolase. Specifically, the IC value of the medium polarity eluate on the inhibition rate of tyrosine diphenolase activity was 0. 50 The value is 0.40 μg / mL, which is significantly better than β-arbutin (which does not have the ability to inhibit tyrosine diphenolase). Combining the results measured by the above two methods, the medium polarity eluate [Example 3] has the best inhibitory effect on in vitro tyrosinase activity.

[0079] The above description is only a preferred embodiment of the present invention, and therefore cannot be used to limit the scope of the present invention. That is, equivalent changes and modifications made according to the patent scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. A method for preparing a natural ingredient of Hericium erinaceus with whitening effect, characterized in that: The preparation method comprises the following steps: (1) crushing the byproducts generated during the processing of Hericium erinaceus fruiting bodies with a pulverizer, sieving to obtain powder, taking an appropriate amount of the powder and soaking it in a 60% by volume ethanol aqueous solution, during which ultrasonic assistance is provided, each ultrasonic temperature is 50 to 55° C., and the ultrasonic time is 30 to 50 min; collecting the filtrate by centrifugation, taking the filtrate and performing rotary evaporation to remove the solvent to obtain a concentrate, taking the concentrate and mixing it with pure water to obtain a concentrated solution; (2) Loading the activated H103 macroporous adsorption resin into a chromatography column, and filling the H103 macroporous adsorption resin with the concentrated solution obtained in step (1) until adsorption is saturated, and initially using pure water to flow through the column to remove non-specifically adsorbed impurities; then using a 20% by volume low-polarity ethanol aqueous solution for elution, collecting an equal volume of low-polarity eluate based on the volume of the elution reagent, and performing rotary evaporation until it becomes a paste, and freeze-drying the paste to obtain a low-polarity eluate, which is a natural ingredient of Hericium erinaceus with whitening effect.

2. The preparation method according to claim 1, characterized in that: The preparation method further comprises the following step (3): (3) After all the low-polarity eluent in step (2) is collected, it is immediately replaced with a 40% by volume medium-polarity ethanol aqueous solution to continue elution. An equal volume of the medium-polarity eluent is collected based on the volume of the elution reagent, and rotary evaporated until it becomes a paste. The paste is freeze-dried to obtain a medium-polarity eluate, which is a natural ingredient of Hericium erinaceus with whitening effect.

3. The preparation method according to claim 2, characterized in that: The preparation method further comprises step (4): (4) After all the intermediate eluent is collected, it is immediately replaced with a 60% high-polarity ethanol aqueous solution to continue elution. An equal volume of high-polarity eluent is collected based on the volume of the elution reagent, and rotary evaporated until it becomes a paste. The paste is freeze-dried to obtain a high-polarity eluate, which is a natural ingredient of Hericium erinaceus with whitening effect.

4. The preparation method according to any one of claims 1 to 3, characterized in that: In step (1), the solid-liquid ratio of the powder to the 60% by volume ethanol aqueous solution is 1 g: (10-20) mL.

5. The preparation method according to any one of claims 1 to 3, characterized in that: In step (1), the concentrate is mixed with pure water in a mass ratio of 1:(3-5).

6. The preparation method according to any one of claims 1 to 3, characterized in that: In step (2), the volume usage of the 20% by volume low-grade ethanol aqueous solution is 2 to 3 times, preferably 2 times, that of the concentrated solution.

7. The preparation method according to claim 2, characterized in that: In step (3), the volume usage of the 40% by volume aqueous solution of medium-polar ethanol is 2 to 3 times, preferably 2 times, that of the concentrated solution.

8. The preparation method according to claim 3, characterized in that: In step (4), the volume usage of the 60% by volume high-polarity ethanol aqueous solution is 2 to 3 times, preferably 2 times, that of the concentrated solution.

9. The preparation method according to any one of claims 1 to 3, characterized in that: The rotary evaporation temperature in step (2), (3) or (4) is 55 to 65°C, preferably 55 to 60°C.

10. A natural ingredient of Hericium erinaceus with whitening effect obtained according to the preparation method of any one of claims 1 to 9.

Images