Preparation method of extract with whitening effect, prepared extract and application thereof

Abstract

The invention discloses a preparation method of an extract with a whitening effect, the prepared extract and application of the extract. The preparation method of the extract with the whitening effect comprises the following steps: (1) crushing a hericium erinaceus sporocarp byproduct by using a crusher, sieving to obtain hericium erinaceus sporocarp byproduct powder, taking a proper amount of powder, soaking with an ethanol aqueous solution with the volume percentage content of 60%, and performing ultrasonic assistance during the period; centrifugally collecting filtrate, combining the filtrate, performing rotary evaporation to obtain a concentrated solution, and adding water to obtain a crude extract; and (2) freeze-drying the crude extract obtained in the step (1) to obtain an alcohol-extracted rotary evaporation substance which is an extract with a whitening effect. The preparation method disclosed by the invention is simple in process step, stable and controllable in process, green and environment-friendly, and beneficial for improving the safety and stability of the product, and the obtained extract has good antioxidant activity and tyrosinase inhibition activity.

Description

Technical Field

[0001] The invention relates to the technical field of cosmetic formulas, and in particular to a method for preparing an extract with whitening effect, the prepared extract with whitening effect and application thereof. Background Art

[0002] Hericium erinaceus belongs to the genus Hericium of the family Hericiaceae, and is mainly distributed in Northeast China, North China, and East China. Hericium erinaceus is not only a delicious edible mushroom, but also a Chinese herbal medicine with important medicinal value. It is rich in terpenoids (2.36 mg / g), flavonoids (0.05 mg / g), polyphenols (0.88 mg / g) and other bioactive substances. Modern medical research shows that Hericium erinaceus extract has significant medicinal value such as anti-oxidation and anti-aging.

[0003] In recent years, with the increase in edible fungi enterprises, more and more by-products are produced during the processing of Hericium erinaceus fruiting bodies. These by-products contain a large number of nutrients such as terpenoids and flavonoids, but most of them are discarded, causing environmental pollution and waste of resources. At the same time, with the rapid development of my country's agriculture and food industry, the requirements for resource utilization and environmental protection are increasing. If the by-products of Hericium erinaceus fruiting body processing can be deeply processed, it can not only reduce the pollution to the environment, but also improve the utilization of active ingredients of Hericium erinaceus fruiting body by-products to a certain extent, and realize the efficient utilization of resources.

[0004] Terpenoids are a class of compounds based on isoprene (C 5 H 8 ) as the basic compound or its derivatives, which are widely found in plants and fungi. Studies have shown that terpenoids have multiple pharmacological activities such as antioxidant, anti-inflammatory and enzyme inhibition. Terpenoids in Hericium erinaceus have been shown to have biological activities such as anti-inflammatory and neuroprotective. Although there are many active ingredients that can be used in whitening products, there are few with practical application value, such as kojic acid and β-arbutin. These inhibitors are not only expensive, but also have certain toxic side effects.

[0005] Studies have shown that terpenoids have been used in whitening products due to their unique whitening effects and pharmacological properties. Patent CN 117771138 A discloses a whitening facial mask of ginseng hydrolysate and its preparation method, wherein methacrylic anhydride-modified dopamine, acrylic acid and dimethylaminoethyl methacrylate are made into hydrogels, and ginsenoside hydrolysates are loaded, so that it has good biocompatibility, antioxidant and whitening effects, but the cost is relatively high. Patent CN 114712369 B discloses a whitening cosmetic active ingredient composition with Gynostemma pentaphyllum secondary saponin as the main active substance, which is safe and reliable for the human body, has no toxic side effects, is non-irritating to the mucous membrane, and has obvious whitening, anti-wrinkle and anti-inflammatory effects, but its preparation method is cumbersome and the whitening effect is not obvious. However, there is no application of Hericium erinaceus fruiting body terpenoids in whitening products. Summary of the invention

[0006] The present invention aims to solve the deficiencies of the prior art and provides a method for preparing an extract with whitening effect, the prepared extract with whitening effect and application thereof. The preparation method has simple process steps, a stable and controllable process, is green and environmentally friendly, helps to improve the safety and stability of the product, and the obtained extract has good antioxidant activity and inhibits tyrosinase activity.

[0007] To achieve this object, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a method for preparing an extract having a whitening effect, comprising the following steps:

[0009] (1) crushing the Hericium erinaceus fruiting body byproduct with a pulverizer, sieving to obtain Hericium erinaceus fruiting body byproduct powder, taking an appropriate amount of the powder and soaking it in a 60% by volume ethanol aqueous solution, during which ultrasonic assistance is used, each ultrasonic temperature is (50-60)°C, and the ultrasonic time is (30-50) min; centrifuging and collecting the filtrate, combining the filtrate and obtaining a concentrate by rotary evaporation, and adding water to obtain a crude extract;

[0010] (2) freeze-drying the crude extract obtained in step (1) to obtain an alcohol-extracted rotary evaporation product.

[0011] Preferably, in step (1), the solid-liquid ratio of Hericium erinaceus fruiting body byproduct powder to ethanol aqueous solution is 1 g: (10-20) mL.

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

[0013] Preferably, the rotary evaporation temperature in step (1) is 55-65°C, more preferably 55-60°C.

[0014] Furthermore, the preparation method further comprises the following steps (3):

[0015] (3) Extracting the crude extract obtained in step (1) with petroleum ether, combining the petroleum ether extracts after sufficient extraction, and rotary evaporating until a paste is formed, and freeze-drying the paste to obtain a petroleum ether extract.

[0016] Preferably, in step (3), the crude extract obtained in step (1) is extracted with an equal volume of petroleum ether, the number of petroleum ether extractions is 3 times, and the petroleum ether extracts are combined after extraction.

[0017] Preferably, the rotary evaporation temperature in step (3) is 55-65°C, more preferably 55-60°C.

[0018] Furthermore, the preparation method further comprises the following steps (4):

[0019] (4) extracting the remaining substance obtained after extraction with petroleum ether in step (3) with ethyl acetate, and obtaining an ethyl acetate extract after sufficient extraction, rotary evaporating the extract until it becomes a paste, and freeze-drying the paste to obtain an ethyl acetate extract.

[0020] Preferably, in step (4), the residual substance obtained after extraction with petroleum ether in step (3) is extracted with an equal volume of ethyl acetate, the number of ethyl acetate extractions is 3, and the ethyl acetate extracts are combined after extraction.

[0021] Preferably, the rotary evaporation temperature in step (4) is 55-65°C, more preferably 55-60°C.

[0022] Furthermore, the preparation method further comprises step (5):

[0023] (5) extracting the residual material obtained after extraction with ethyl acetate in step (4) with water-saturated n-butanol to obtain a water-saturated n-butanol extract after sufficient extraction, rotary evaporating until it becomes a paste, and freeze-drying the paste to obtain a water-saturated n-butanol extract.

[0024] Preferably, in step (5), the aqueous layer obtained after extraction with ethyl acetate in step (4) is extracted with water-saturated n-butanol for 3 times, and the water-saturated n-butanol extracts are combined after extraction.

[0025] Preferably, the rotary evaporation temperature in step (5) is 55-65°C, more preferably 55-60°C.

[0026] Furthermore, the preparation method further comprises step (6):

[0027] (6) The residual material obtained after extraction with water-saturated n-butanol in step (5) is subjected to rotary evaporation until it becomes a paste, and the paste is freeze-dried to obtain an aqueous layer extract.

[0028] Preferably, the rotary evaporation temperature in step (6) is 55-65°C, more preferably 55-60°C.

[0029] In the present invention, the alcohol-extracted rotary evaporation product, petroleum ether extract, ethyl acetate extract, water-saturated n-butanol extract and aqueous layer extract are all extracts with whitening effect.

[0030] In a second aspect, the present invention provides an extract with whitening effect obtained according to the preparation method described in the first aspect.

[0031] In a third aspect, the present invention provides the use of the extract with whitening effect according to the second aspect as an active ingredient of whitening cosmetics.

[0032] Compared with the background technology, this technical solution has the following advantages:

[0033] (1) The present invention extracts the by-products of Hericium erinaceus fruiting bodies with ethanol-water solution, and then successively screens them using solvents of different polarities to obtain a variety of Hericium erinaceus fruiting body by-product extracts. The antioxidant activity (especially the DPPH free radical scavenging effect) and tyrosinase inhibition activity of these extracts are better than β-arbutin, and can be used as active ingredients in whitening cosmetics.

[0034] (2) The aqueous layer extract of the Hericium erinaceus fruiting body byproduct obtained by the present invention not only shows significant advantages in antioxidant activity and inhibition of tyrosinase activity, but also can significantly inhibit melanin deposition.

[0035] (3) The present invention utilizes the by-products of Hericium erinaceus fruiting bodies, has low production cost and high added value, is beneficial to efficient utilization of resources and environmental protection, and at the same time improves the economic value of the product. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0037] Figure 1 This is the separation process of extracts of different polarities from Hericium erinaceus fruiting bodies.

[0038] Figure 2 The scavenging ability of the products of Examples 1-5 and β-arbutin [positive control] on DPPH free radicals.

[0039] Figure 3 The scavenging ability of the products of Examples 1-5 and β-arbutin [positive control] on ABTS free radicals.

[0040] Figure 4 The inhibitory effects of the products of Examples 1-5 and β-arbutin [positive control] on tyrosine monophenolase activity.

[0041] Figure 5 The inhibitory effects of the products of Examples 1-5 and β-arbutin [positive control] on tyrosine diphenolase activity.

[0042] Figure 6 The effects of the products of Examples 1-5 and β-arbutin [positive control] on the viability of B16-F10 mouse melanoma cells.

[0043] Figure 7 The relative melanin content of the products of Examples 1-5 and β-arbutin [positive control] in B16-F10 mouse melanoma cells. DETAILED DESCRIPTION

[0044] 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.

[0045] Examples 1-5

[0046] See also Figure 1 , a method for preparing an extract with whitening effect, the specific steps are as follows:

[0047] Step 1. 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 an 80-mesh sieve and weigh an appropriate amount of powder.

[0048] Step 2. Ethanol soaking and ultrasonic-assisted 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 1g:15mL. Ultrasonic assistance is performed during the soaking process, and each ultrasonic temperature is 55°C and the ultrasonic time is 30min.

[0049] Step 3. Collect the filtrate by centrifugation: Centrifuge the soaked solution at a centrifugal rate of 12000 r / min for 20 min. Collect the filtrate, combine to obtain 900 mL of filtrate, and concentrate by rotary evaporation (the temperature of the constant temperature water bath is 60°C, the time is 60 min, the speed is 80 rpm, and the vacuum degree is -0.1 MPa), and add water at a solid-liquid ratio of V:V=1:3 to obtain a crude extract.

[0050] Step 4. Alcohol extraction and rotary evaporation: freeze-dry part of the crude extract in step 3 (cold trap temperature is -80°C, freezing chamber temperature is -45°C, vacuum degree is 6.8 Pa, and freeze-drying time is 2 days) to obtain alcohol extraction and rotary evaporation of Hericium erinaceus fruiting body [Example 1].

[0051] Step 5. Petroleum ether extract: The remaining crude extract in step 3 was extracted with an equal volume of petroleum ether for 3 times, and the petroleum ether extracts were combined and rotary evaporated (the temperature of the constant temperature water tank was 60°C, the time was 25 min, the speed was 80 rpm, and the vacuum degree was -0.1 MPa) until it became a paste. The paste was freeze-dried (the cold trap temperature was -80°C, the freezing chamber temperature was -45°C, the vacuum degree was 6.8 Pa, and the freeze-drying time was 2 days) to obtain the petroleum ether extract of Hericium erinaceus fruiting body [Example 2].

[0052] Step 6. Ethyl acetate extract: The residual material obtained after extraction with petroleum ether in step 5 was extracted with an equal volume of ethyl acetate, extracted 3 times, and the ethyl acetate extracts were combined and rotary evaporated (the temperature of the constant temperature water tank was 60°C, the time was 35min, the speed was 80rpm, and the vacuum degree was -0.1MPa) until it became a paste. The paste was freeze-dried (the cold trap temperature was -80°C, the freezing chamber temperature was -45°C, the vacuum degree was 6.8Pa, and the freeze-drying time was 2 days) to obtain the ethyl acetate extract of Hericium erinaceus fruiting body [Example 3].

[0053] Step 7. Water-saturated n-butanol extract: The residual material obtained after extraction with ethyl acetate in step 6 was extracted with an equal volume of water-saturated n-butanol for 3 times. The water-saturated n-butanol extracts were combined and evaporated by rotary evaporation (the temperature of the constant temperature water tank was 65°C, the time was 75min, the speed was 80rpm, and the vacuum degree was -0.1MPa) until it was a paste. The paste was freeze-dried (the cold trap temperature was -80°C, the freezing chamber temperature was -45°C, the vacuum degree was 6.8Pa, and the freeze-drying time was 2 days) to obtain the water-saturated n-butanol extract of Hericium erinaceus fruiting body [Example 4].

[0054] Step 8. Aqueous layer extract: After the above-mentioned petroleum ether, ethyl acetate and water-saturated n-butanol are extracted step by step, the remaining substances have greater polarity and are not extracted by other organic solvents, so these remaining substances are named aqueous layer extracts. The aqueous layer extract is evaporated by rotary evaporation (the temperature of the constant temperature water tank is 65°C, the time is 45min, the speed is 80rpm, and the vacuum degree is -0.1MPa) until it becomes a paste. The paste is freeze-dried (the cold trap temperature is -80°C, the freezing chamber temperature is -45°C, the vacuum degree is 6.8Pa, and the freeze-drying time is 2 days) to obtain the aqueous layer extract of Hericium erinaceus fruiting body [Example 5].

[0055] The extract prepared by the above steps of the present invention has good antioxidant activity and tyrosinase inhibition activity, can significantly inhibit B16-F10 melanin deposition, and is suitable for the preparation of whitening products and other related applications.

[0056] Embodiment 6:

[0057] (1) The products of Examples 1-5 were respectively dissolved in phosphate buffered saline (PBS): dimethyl sulfoxide (DMSO) (9:1, v / v) for subsequent application research. The absorbance at 547 nm was determined according to the glacial acetic acid 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 1-5. The sample solutions to be tested were obtained by diluting the terpenoid compounds in the products of Examples 1-5 according to the concentration gradient.

[0058] (2) Determination of the DPPH free radical scavenging ability of the products of Examples 1-5: First, prepare a 0.05 mg / mL DPPH ethanol solution; 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.

[0059] (3) Determination of ABTS free radical scavenging ability of the products of Examples 1-5: 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 a detection value of about 0.7 at 734 nm on an ELISA reader; 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.

[0060] (4) Determination of the ability of the products of Examples 1-5 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 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-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.

[0061] (5) Determination of the in vitro tyrosine diphenolase inhibitory ability of the products of Examples 1-5: 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. Next, 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 fully react at 37°C for 10 min, and the absorbance was measured at 475 nm.

[0062] (6) CCK8 method was used to determine the effect of the products of Example 1-5 on the viability of B16-F10 cells: First, a complete culture medium was prepared according to a volume ratio of DMEM low sodium bicarbonate culture medium: fetal bovine serum of 9:1. B16-F10 cells in the logarithmic growth phase were taken, and the cell count was performed to adjust the cell concentration to 4×10 3 / well were inoculated into 96-well plates and incubated in 5% CO 2, and cultured overnight in a 37°C constant temperature incubator. The next day, after the cells adhered, the culture medium was removed, and 100uL of the sample solution to be tested in step (1) was added to each well, and the culture was continued for 48h. After the culture was completed, each well was washed three times with PBS, and a culture medium containing 10% CCK-8 was added at 120μL / well, and 5% CO 2 , incubate in a 37°C constant temperature incubator for 2 h. Take 100 μL / well of the supernatant and transfer it to a new 96-well plate, and measure the absorbance at 450 nm with an ELISA reader.

[0063] (7) Determination of the effect of the product of Example 1-5 on the melanin inhibition of B16-F10 mouse melanoma cells: B16-F10 cells in the logarithmic growth phase were taken, and the cell concentration was adjusted to 5×10 5 / dish was inoculated into 100 mm culture dishes and incubated in 5% CO 2 , and cultured overnight in a 37°C constant temperature incubator. Select specific components and concentration samples based on the cytotoxicity results, add 10 mL of the sample solution to each dish, and test the melanin content after 48 hours of culture. Aspirate the supernatant, wash once with PBS solution, collect the cells in a centrifuge tube after trypsin digestion, centrifuge at 1500 rpm for 10 minutes, and discard the supernatant. Add 1 mL of 1 mol·L containing 10% DMSO -1 NaOH solution, incubated in a water bath at 80°C for 60 min, and then transferred to 96-well culture plates, and the absorbance at 475 nm was detected by a microplate reader. The calculation formula for melanin content is as follows: melanin relative content (%) = (OD value of experimental well - OD value of blank well) / (OD value of control well - OD value of blank well) × 100%.

[0064] Comparative Example 1:

[0065] Take β-arbutin as positive control: accurately weigh 200 mg of β-arbutin, and dilute to volume with 100 mL PBS: DMSO (9:1, v / v) to obtain 2 mg / mL β-arbutin [positive control], and gradient dilute to a specific concentration to obtain β-arbutin [positive control] [Comparative Example 1]. Refer to Example 6, except that the experimental sample is replaced with β-arbutin [positive control], and the effect of the sample on the viability of B16-F10 cells at different concentrations.

[0066] Test results

[0067] In vitro antioxidant activity test: The antioxidant activity of the gradient concentration samples prepared in Example 6 and Comparative Example 1 was studied, and the results are as follows: Figure 2 and Figure 3The results of the DPPH method and the ABTS method are both expressed in terms of antioxidant rate (%). Combining the results of the two methods, it can be seen that the antioxidant activity of different extracts of Hericium erinaceus fruiting bodies determined by the DPPH method is in the following order from large to small: aqueous layer extract > petroleum ether extract > ethyl acetate extract > water-saturated n-butanol extract > alcohol rotary evaporation liquid > β-arbutin [positive control]. Among them, the aqueous layer extract of Hericium erinaceus fruiting body has the highest antioxidant rate among these extracts, and the water-saturated n-butanol extract has the lowest antioxidant rate. In addition, the DPPH self- The free radical scavenging effect of Hericium erinaceus fruiting body is better than that of β-arbutin [positive control]; the antioxidant activity of different extracts of Hericium erinaceus fruiting body determined by ABTS method is in the following order from small to large: β-arbutin [positive control] > water-saturated n-butanol extract > water layer extract > petroleum ether extract > ethyl acetate extract > alcohol extraction rotary distillation liquid, among which, the water-saturated n-butanol extract of Hericium erinaceus fruiting body has the highest antioxidant rate among these extracts, while the ethyl acetate extract of Hericium erinaceus fruiting body has the lowest antioxidant rate, and the ABTS free radical scavenging effect of all extracts is better than that of alcohol extraction rotary distillation liquid. Combining the results measured by the above two methods, the water layer extract of Hericium erinaceus fruiting body has the best antioxidant effect.

[0068] In vitro tyrosinase activity inhibition: The gradient concentration samples prepared in Example 6 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 phenolase and tyrosine diphenolase are both expressed in terms of enzyme activity inhibition rate (%). Combining the results of the above two methods, it can be seen that for the results of in vitro inhibition of tyrosine phenolase, the inhibition effects of different extracts from Hericium erinaceus fruiting bodies on tyrosine phenolase activity are in the following order from large to small: ethyl acetate extract > alcohol rotary evaporation liquid > water layer extract > water-saturated n-butanol extract > petroleum ether extract > β-arbutin [positive control]. Among them, the ethyl acetate extract of Hericium erinaceus fruiting body has the highest inhibition rate on tyrosine phenolase among these extracts, and the petroleum ether extract The inhibitory effect of all extracts on tyrosine diphenolase activity was the smallest, and the inhibitory effect of all extracts on tyrosine monophenolase activity was better than that of β-arbutin [positive control]; the inhibitory effect of different extracts of Hericium erinaceus fruiting body on tyrosine diphenolase activity in vitro was measured in the following order from large to small: aqueous layer extract > water-saturated n-butanol extract > ethyl acetate extract > alcohol extraction rotary evaporation liquid > petroleum ether extract, among which, the inhibitory rate of tyrosine diphenolase of the aqueous layer extract of Hericium erinaceus fruiting body was the largest among these extracts, while the petroleum ether extract of Hericium erinaceus fruiting body was the smallest, and β-arbutin [positive control] did not have an inhibitory effect on tyrosinase activity. Combining the results measured by the above two methods, the inhibitory effect of the aqueous layer extract of Hericium erinaceus fruiting body on tyrosinase activity in vitro was the best.

[0069] B16-F10 cytotoxicity experiment: The gradient concentration samples prepared in Example 6 and Comparative Example 1 were tested for their effects on B16-F10 cell viability. The results are as follows: Figure 6 As shown. Petroleum ether extract, ethyl acetate extract, n-butanol extract, water layer extract, alcohol extraction rotary evaporation liquid and β-arbutin (positive control) have different degrees of inhibitory effect on cell viability, and this inhibitory effect becomes more obvious with the increase of concentration. The cytotoxic effects of different extracts of Hericium erinaceus fruiting bodies on B16-F10 cells are measured in the following order from large to small: petroleum ether extract > ethyl acetate extract > alcohol extraction rotary evaporation liquid > water-saturated n-butanol extract > water layer extract > β-arbutin (positive control). Among them, when the concentration is lower than 150μg / mL, after the water layer extract acts on B16-F10 cells, the cell survival rate is above 60%, and most of the cells are in a normal growth state through observation under a microscope, indicating that when the concentration of the water layer extract of Hericium erinaceus fruiting body is lower than 150μg / mL, the toxic effect on B16-F10 cells is small.

[0070] B16-F10 melanin deposition inhibition experiment: The gradient concentration samples prepared in Example 6 and Comparative Example 1 were used to test the effect of inhibiting melanin deposition in B16-F10 cells. The results are shown in the figure. Figure 7 As shown. The aqueous layer extract has a significant inhibitory effect on melanin deposition. When its concentration is 100μg / mL, the cell survival rate is about 70%, and the relative content of melanin is less than about 50%. When the concentration of the alcohol extraction rotary evaporation liquid is 10μg / mL, the cell survival rate is also about 70%, and the relative content of melanin is about 80%. Therefore, it is speculated that the different extracts of Hericium erinaceus fruiting body have poor melanin deposition effects, but the aqueous layer extract has a certain inhibitory activity on melanin deposition.

[0071] 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 an extract having a whitening effect, characterized in that: The preparation method comprises the following steps: (1) crushing the Hericium erinaceus fruiting body byproduct with a pulverizer, sieving to obtain Hericium erinaceus fruiting body byproduct powder, taking an appropriate amount of the powder and soaking it in a 60% by volume ethanol aqueous solution, during which ultrasonic assistance is used, each ultrasonic temperature is (50-60)°C, and the ultrasonic time is (30-50) min; centrifuging and collecting the filtrate, combining the filtrate and obtaining a concentrate by rotary evaporation, and adding water to obtain a crude extract; (2) freeze-drying the crude extract obtained in step (1) to obtain an alcohol-extracted rotary evaporation product, wherein the alcohol-extracted rotary evaporation product is an extract with whitening effect.

2. The preparation method according to claim 1, characterized in that: In step (1), the solid-liquid ratio of Hericium erinaceus fruiting body byproduct powder to ethanol aqueous solution is 1 g: (10-20) mL.

3. The preparation method according to claim 1 or 2, characterized in that: The preparation method further comprises the following steps (3): (3) Extracting the crude extract obtained in step (1) with petroleum ether, combining the petroleum ether extracts after sufficient extraction, and performing rotary evaporation until the paste is formed, freeze-drying the paste to obtain a petroleum ether extract, wherein the petroleum ether extract is an extract having a whitening effect.

4. The preparation method according to claim 3, characterized in that: The preparation method further comprises the following steps (4): (4) extracting the remaining substance obtained after extraction with petroleum ether in step (3) with ethyl acetate, obtaining an ethyl acetate extract after sufficient extraction, performing rotary evaporation until it becomes a paste, freeze-drying the paste, and obtaining an ethyl acetate extract, wherein the ethyl acetate extract is an extract having a whitening effect.

5. The preparation method according to claim 4, characterized in that: The preparation method further comprises step (5): (5) extracting the remaining substance obtained after extraction with ethyl acetate in step (4) with water-saturated n-butanol to obtain a water-saturated n-butanol extract after sufficient extraction, performing rotary evaporation until it becomes a paste, freeze-drying the paste, and obtaining a water-saturated n-butanol extract, wherein the water-saturated n-butanol extract is an extract having a whitening effect.

6. The preparation method according to claim 5, characterized in that: The preparation method further comprises step (6): (6) The residual material obtained after extraction with water-saturated n-butanol in step (5) is subjected to rotary evaporation until it becomes a paste, and the paste is freeze-dried to obtain an aqueous layer extract, wherein the aqueous layer extract is an extract having a whitening effect.

7. The preparation method according to any one of claims 3 to 6, characterized in that: In step (3), the crude extract obtained in step (1) is extracted with an equal volume of petroleum ether, the number of petroleum ether extractions is 3 times, and the petroleum ether extracts are combined after extraction; In step (4), the residual substance obtained after extraction with petroleum ether in step (3) is extracted with an equal volume of ethyl acetate, the number of extractions with ethyl acetate is 3 times, and the ethyl acetate extracts are combined after extraction; In step (5), the aqueous layer obtained after extraction with ethyl acetate in step (4) is extracted with water-saturated n-butanol for 3 times, and the water-saturated n-butanol extracts are combined after extraction.

8. The preparation method according to any one of claims 1 to 6, characterized in that: The preparation method according to any one of claims 3 to 6, characterized in that the rotary evaporation temperature in steps (1), (3), (4), (5) or (6) is 55 to 65°C.

9. An extract with whitening effect obtained according to the preparation method according to any one of claims 1 to 8.

10. Use of the extract with whitening effect as claimed in claim 9 as an active ingredient of whitening cosmetics.

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