Preparation method, application and product of anti-aging abalone extract

Abstract

The invention discloses a preparation method of an anti-aging abalone extract, application of the anti-aging abalone extract and a product, and the preparation method is characterized by comprising the following steps: uniformly mixing abalone powder with low-concentration ethanol to obtain a mixture, and carrying out enzymolysis on the mixture to obtain abalone enzymatic hydrolysate; separating and purifying the obtained abalone enzymatic hydrolysate to obtain enriched liquid; and concentrating the obtained enriched solution to obtain a concentrated solution. According to the method, a low-concentration ethanol enzymolysis technology is creatively used, and the abalone powder is subjected to enzymolysis in a low-concentration ethanol solution by utilizing alcohol-resistant neutral protease, so that effective active ingredients can be released, and impurities can be removed, thereby facilitating enrichment and purification of the active ingredients. According to the preparation method provided by the invention, the cost is reduced, the working section is reduced, and meanwhile, the preparation method is safer due to low-concentration ethanol and no need of adding an acid-base reagent in the working section to adjust the pH value.

Description

Technical Field

[0001] The present invention relates to the field of biological extracts, and in particular to a preparation method of an anti-aging abalone extract and its application and product. Background Art

[0002] Physiological aging is a series of natural and complex physiological changes that occur as the body ages. It is a gradual process. As time goes by, the physiological functions of the human body gradually decline. In addition to the epigenetic characteristics, human aging is also closely related to oxidative stress response, excessive cell apoptosis, telomere shortening, gene expression disorders, mitochondrial dysfunction, DNA damage accumulation, reduced protease activity, impaired immune function, changes in intercellular communication, inflammatory response, etc., and anti-oxidation is one of the most basic functions of anti-aging products.

[0003] Abalone, also known as mirror fish, nine-hole snail, eye-catching fish, and general cap, has more than 100 species of abalone in the world. It is widely loved for its rich nutritional value and delicious taste. It is known as "a mouthful of abalone is a mouthful of gold". According to analysis, the edible part of abalone contains 24% protein, 0.44% fat, and contains a variety of vitamins and some special trace components. It has high therapeutic effects and medicinal value, such as supplementing the protein and other nutrients needed by the human body. The active substances extracted from it also have the effects of anti-oxidation, enhancing immunity, protecting the liver and sobering up. The current methods for extracting active substances from abalone are as follows:

[0004] The invention patent with application number 201510626266.1 discloses a method for preparing abalone viscera antioxidant: using abalone viscera as raw material, abalone viscera antioxidant peptide powder is obtained through defatting, protein removal, hot water extraction, high-speed centrifugation and dialysis, and then mixed with sodium alginate and added to calcium chloride, and finally vacuum dried to obtain a safe, non-toxic, stable, and strong antioxidant activity abalone viscera protein peptide antioxidant. However, the production process of this method is complicated, and the product is mainly used for meat preservation and is not suitable for direct consumption as food.

[0005] The invention patent with application number 202010299549.0 discloses an abalone viscera extract and its application. The method comprises homogenizing, freeze-drying, ultrasound-assisted extraction, constant temperature extraction, centrifugation, reduced pressure concentration, and freeze-drying the abalone viscera to obtain an abalone viscera extract that protects the reproductive system of female mice. However, this method requires two freeze-drying steps, which is costly and not suitable for large-scale industrial production.

[0006] The invention patent with application number 202010740272.0 discloses an industrial abalone peptide extraction method, which is to make abalone into minced meat, add alkali to defat, wash, adjust pH, extract, separate oil, enzymatically hydrolyze, inactivate enzymes, deodorize and decolorize, nanofilter, concentrate, and spray dry to obtain abalone peptides. Among the abalone peptides obtained by this method, oligopeptides with a molecular weight of less than 1000Da account for 86%. Although the degree of enzymatic hydrolysis is high, the efficacy of the obtained small molecule abalone peptides has not been disclosed.

[0007] The invention patent with application number 202310677452.2 discloses a uric acid-lowering abalone visceral polypeptide and its preparation method and application. The method uses abalone viscera as raw material, and obtains a uric acid-lowering abalone visceral polypeptide through homogenization, enzymatic hydrolysis under weak alkaline conditions, enzymatic hydrolysis under acidic conditions, centrifugation, ultrafiltration, and chromatography, and its amino acid sequence is LES. This method requires the pH to be adjusted to 1.0-3.0 during enzymatic hydrolysis under acidic conditions, which is not suitable for industrial production.

[0008] The existing technology for extracting abalone active substances is relatively complex and costly, and the application of abalone extracts is limited. Therefore, there is an urgent need for a low-cost, simple process preparation method for abalone extracts, and further exploration of the application of abalone extracts. Summary of the invention

[0009] The purpose of the present invention is to provide a simple, safe and industrially producible method for preparing an abalone extract with anti-aging efficacy and a product.

[0010] To achieve the above object, the first aspect of the present invention provides a method for preparing an anti-aging abalone extract, characterized in that it comprises:

[0011] S1, abalone powder and low concentration of ethanol are mixed to obtain a mixture, and the mixture is enzymolyzed to obtain an abalone hydrolyzate;

[0012] S2, separating and purifying the abalone hydrolysate obtained in S1 to obtain an enriched solution;

[0013] S3, concentrating the enriched solution obtained in S2 to obtain a concentrated solution.

[0014] The low-concentration ethanol described in the present invention is ethanol with a volume percentage of less than 40%, and the ethanol concentration is preferably 20-30% by volume.

[0015] Furthermore, the concentrated solution obtained in step S3 is further spray-dried to obtain abalone extract powder.

[0016] Furthermore, the enzymolysis in step S1 is performed by a neutral protease that is active in low-concentration ethanol; preferably, the low-concentration ethanol in step S1 is food-grade ethanol with a volume percentage of 20-30%. The neutral protease maintains 50%-70% of the original enzyme activity in 20%-30% ethanol, and the enzymolysis of abalone powder can release effective active ingredients. Ethanol can also be used to precipitate some soluble impurities so that impurities can be removed during separation and purification, thereby enriching and purifying active ingredients, so as to achieve the purpose of reducing steps.

[0017] Furthermore, in step S1, the mass ratio of abalone powder: low concentration ethanol: neutral protease is 1:3-5:0.005-0.01.

[0018] Furthermore, the abalone powder in step S1 is prepared by pre-treating abalone, drying and crushing the edible part of abalone to obtain abalone powder;

[0019] Preferably, the pretreatment is to shell the live abalone, take out the meat and viscera, and further wash, blanch, cool and drain to obtain the edible part of the abalone;

[0020] Preferably, the drying temperature is 45-55°C;

[0021] Preferably, the edible part of the abalone is dried until it can be crushed into powder; more preferably, the drying time is 40-48 hours;

[0022] Blanching can kill some spoilage bacteria and endogenous enzymes, and washing again with normal temperature water can quickly cool down the temperature, preventing the central temperature from being high during drying, thereby preventing the raw materials from deteriorating. At the same time, drying the edible part of abalone with a dryer can speed up the drying rate compared to sun exposure, preventing the edible part of abalone from deteriorating during long-term sun exposure, and drying at a lower temperature can also prevent the edible part of abalone from maturing. After drying the edible part of abalone, crushing it into abalone powder can allow the enzyme to fully contact with the protein, thereby improving the enzymolysis efficiency and product yield.

[0023] Furthermore, the temperature of enzymolysis in step S1 is 50-55° C.; preferably, the time of enzymolysis in step S2 is 1.5 h-2.5 h.

[0024] Furthermore, in step S2, the abalone hydrolysate is centrifuged to obtain a supernatant, the supernatant is coarsely filtered to obtain a filtrate, the filtrate is ultrafiltered to obtain a permeate, and the permeate is nanofiltered to obtain an enriched solution containing active ingredients;

[0025] Preferably, the centrifugation is performed at 10000-12000 rpm for 25-30 min;

[0026] Preferably, the coarse filtration is performed using a filter cloth with a mesh size of 150-250;

[0027] Preferably, the ultrafiltration is to filter the filtrate using an ultrafiltration membrane separation device with a molecular weight cutoff of 3000Da;

[0028] Preferably, the nanofiltration is to use a nanofiltration membrane separation device with a molecular weight cutoff of 500Da to intercept the permeate.

[0029] Experiments have found that components with a molecular weight of 500-3000Da in abalone hydrolysate have better anti-aging effects than other components. Therefore, in production, precipitation and macromolecular substances are first removed by centrifugation and coarse filtration to prevent ultrafiltration membrane separation equipment from being easily blocked, slowing down the rate of flux decline of the membrane equipment and thus improving the efficiency of separation and purification; the filtrate is then subjected to ultrafiltration to remove substances with a molecular weight of >3000Da in the filtrate to obtain a permeate with a high content of active ingredients; the permeate is then subjected to nanofiltration to remove substances with a molecular weight of <500Da in the permeate, and substances with a molecular weight of >500Da in the permeate are retained to enrich the permeate to obtain an enriched solution containing a higher content of active ingredients.

[0030] Further, the concentration of the concentrated solution is 15-20°Bx; preferably, the enriched solution in step S3 is concentrated at 50-60°C to obtain the concentrated solution;

[0031] The abalone extract prepared by the preparation method provided in the first aspect of the present invention has anti-aging application.

[0032] The second aspect of the present invention provides a product containing an anti-aging abalone extract, wherein the abalone extract obtained by the preparation method provided in the first aspect of the present invention is prepared into one or more of capsules, tablets, granules, oral liquids or beverages.

[0033] The preparation method proposed in the present invention prepares abalone powder from pretreated abalone and mixes it with low-concentration ethanol, and enzymolyzes the abalone powder with neutral protease at natural pH, thereby reducing the introduction of ash and making the purity of the effective ingredients in the product higher.

[0034] The enzymatic hydrolysis and alcohol precipitation extraction in the prior art are combined together, and a neutral protease having 50%-70% of the original enzyme activity in ethanol is used to enzymatically hydrolyze abalone powder, and impurities can be removed while releasing effective active ingredients. The obtained extract has higher antioxidant activity than that of enzymatic hydrolysis without ethanol, and in subsequent treatment, only the abalone enzymatic hydrolyzate needs to be purified and concentrated to obtain an abalone extract containing active ingredients. The prepared abalone extract can be eaten directly without reprocessing, which reduces costs and saves work sections. At the same time, the low concentration of ethanol and the absence of adding acid-base reagents to adjust pH in the work section also make the preparation method proposed in the present invention safer.

[0035] The concentrate can be further spray-dried to prepare powdered abalone extract for storage, transportation and further processing. At the same time, in order to better adapt to the consumer market, the abalone extract containing active ingredients can be prepared into capsules, tablets, granules, oral liquids or beverages to cater to the preferences of different consumers.

[0036] The present invention creatively uses low-concentration ethanol enzymatic hydrolysis technology, and utilizes alcohol-resistant neutral protease to enzymatically hydrolyze abalone powder in a low-concentration ethanol solution, which can not only release effective active ingredients, but also utilize ethanol to precipitate some soluble impurities, making it easier to enrich the active ingredients through subsequent separation and purification, thereby specifically obtaining an abalone extract rich in active ingredients.

[0037] In vitro experiments have found that when the concentration of the abalone extract powder obtained by the preparation method proposed in the present invention is 10 mg / mL, the scavenging rate of the free radical DPPH· is 77.5%-87.4%. Therefore, the abalone extract of the present invention has anti-aging uses. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The figure is a graph showing the determination result of DPPH clearance rate of components with different molecular weights in Example 1 of the present invention;

[0039] Figure 2 This is a graph showing the measurement results of the DPPH scavenging effect of the products containing the anti-aging abalone extract in Example 1, Comparative Example 1, Comparative Example 2, and Comparative Example 3 of the present invention. DETAILED DESCRIPTION

[0040] Embodiments of the present invention are described in detail below, and the examples of the embodiments are intended to be used to explain the present invention, and should not be construed as limiting the present invention. In the embodiments, if specific techniques or conditions are not indicated, the techniques or conditions described in the literature in this area or the product specifications are carried out. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained commercially. In the following embodiments, if not clearly stated, "%" refers to weight percentage.

[0041] 1. The neutral protease used in the present invention is a neutral protease produced by Nanning Pangbo Bioengineering Co., Ltd., and the enzyme activity is 150000U / g.

[0042] 2. The abalone used in the embodiment of the present invention is any one of Haliotis discus hannai, Haliotis discus, Haliotis discus, Haliotis discus, and Haliotis discus that are fresh and have no obvious fishy smell.

[0043] 3. The preparation method of the anti-aging abalone extract of the present invention is as follows:

[0044] S1, abalone powder and low concentration of ethanol are mixed to obtain mixture, and the mixture is hydrolyzed to obtain abalone hydrolyzate;

[0045] S2, separating and purifying the abalone hydrolysate obtained in S1 to obtain an enriched solution;

[0046] S3, concentrating the enriched solution obtained in S2 to obtain a concentrated solution.

[0047] Preferably, the concentrated solution obtained in step S3 is further spray-dried to obtain abalone extract powder.

[0048] Preferably, the enzymatic hydrolysis in step S1 is performed by a neutral protease that is active in low concentration ethanol; more preferably, the ethanol in step S1 is food grade ethanol with a volume percentage of 20-30%.

[0049] Preferably, in step S1, the mass ratio of abalone powder: low concentration ethanol: neutral protease is 1:3-5:0.005-0.01.

[0050] Preferably, the abalone powder in step S1 is obtained by pre-treating abalone, drying and crushing the edible part of abalone to obtain abalone powder;

[0051] More preferably, the pretreatment is to shell the live abalone, take out the meat and viscera, and further wash, blanch, cool and drain to obtain the edible part of the abalone;

[0052] More preferably, the drying temperature is 45-55°C;

[0053] More preferably, the edible part of the abalone is dried until it can be crushed into powder; more preferably, the drying time is 40-48 hours.

[0054] Preferably, the temperature of enzymolysis in step S1 is 50-55° C.; more preferably, the time of enzymolysis in step S2 is 1.5 h-2.5 h.

[0055] Preferably, step S2 is to centrifuge the abalone hydrolysate to obtain a supernatant, perform coarse filtration on the supernatant to obtain a filtrate, perform ultrafiltration on the filtrate to obtain a permeate, and perform nanofiltration on the permeate to obtain an enriched solution rich in active ingredients;

[0056] More preferably, the centrifugation is performed at 10000-12000 rpm for 25-30 min;

[0057] More preferably, the coarse filtration is performed using a filter cloth with a mesh size of 150-250;

[0058] More preferably, the ultrafiltration is to filter the filtrate using an ultrafiltration membrane separation device with a molecular weight cutoff of 3000Da;

[0059] More preferably, the nanofiltration is to use a nanofiltration membrane separation device with a molecular weight cutoff of 500Da to intercept the permeate.

[0060] Preferably, the concentration of the concentrated solution is 15-20° Bx; preferably, the enriched solution in step S3 is concentrated at 50-60° C. to obtain the concentrated solution.

[0061] Preferably, the concentrated solution obtained in step S3 is further spray-dried to obtain abalone extract powder.

[0062] Embodiment 1:

[0063] Take 50 kg of fresh abalone raw material, remove the shell, take out the meat and viscera, wash once with normal temperature water, blanch with boiling water for 5 minutes, wash once with normal temperature water, drain, dry at 50 ° C for 48 hours, crush with an ultrafine grinder and sieve to obtain abalone powder with a particle size of less than 120 mesh.

[0064] S1, add 25% food-grade ethanol aqueous solution with a volume percentage of 5 times its weight to abalone powder, after mixing, warm up to 55 DEG C, add 500g neutral protease, enzymolysis 2h under natural pH condition, constantly stir during enzymolysis, enzymolysis ends, and obtains abalone enzymolysis solution.

[0065] S2, abalone enzymolysis liquid is centrifuged at 12000rpm for 30min, supernatant is taken, filtrate is obtained after coarse filtration with 200 mesh filter cloths, then filtrate is carried out ultrafiltration by the ultrafiltration membrane separation device that molecular weight cutoff is 3000Da to obtain permeate, then permeate is carried out nanofiltration by the nano-membrane separation device that molecular weight cutoff is 500Da to obtain enriched liquid.

[0066] S3. The obtained enriched liquid is passed into a vacuum concentration tank, and concentrated under reduced pressure to 15°Bx at 60°C to obtain a concentrated liquid.

[0067] The concentrated solution is spray dried at 180°C to make powder, and then packaged to obtain a product containing anti-aging abalone extract. Test Example 1:

[0068] Test on the scavenging effect of components with different molecular weights on DPPH·:

[0069] Pretreatment: The filtrate (obtained by coarse filtration through filter cloth), the retentate (components with a molecular weight of >3000Da obtained by ultrafiltration), the enriched solution (components with a molecular weight between 500-3000Da obtained by nanofiltration) and the waste liquid (components with a molecular weight of <500Da obtained by nanofiltration) in Example 1 were freeze-dried and then ground to obtain samples to be tested.

[0070] Preparation of test solution: Take 1.00g of the sample to be tested and dissolve it in 100mL of deionized water to prepare a test solution with a concentration of 10mg / mL.

[0071] Add 500μL of the test solution and 500μL of 0.2mmol / L DPPH·anhydrous ethanol solution to a 2.0mL centrifuge tube, mix thoroughly and react for 30 minutes in the dark. After the reaction, centrifuge at 5000rpm for 10 minutes, take the supernatant and measure the absorbance at 517nm. The control group uses deionized water instead of the sample, the blank group uses anhydrous ethanol instead of DPPH anhydrous ethanol solution, and Vc is used as a positive control.

[0072] The calculation method of DPPH clearance rate is as follows:

[0073] DPPH·scavenging rate (%) = [1-(A 样 -A 空 ) / A 对 ]×100

[0074] Where: A 样 is the absorbance value measured for the sample group;

[0075] A 空 is the absorbance value measured for the blank group;

[0076] A 对 This is the absorbance value measured for the control group.

[0077] The results of DPPH clearance were as follows: Figure 1 As shown:

[0078] Among the components with different molecular weights, the enriched solution obtained by nanofiltration (molecular weight range of 500-3000Da) has the highest DPPH clearance rate, so this component is selected for the preparation of anti-aging products.

[0079] Comparative Example 1:

[0080] Take 50 kg of fresh abalone raw material, remove the shell, take out the meat and viscera, wash once with normal temperature water, blanch with boiling water for 5 minutes, wash once with normal temperature water, drain, dry at 50 ° C for 48 hours, crush with an ultrafine grinder and sieve to obtain abalone powder with a particle size of less than 120 mesh.

[0081] S1, in abalone powder, add 5 times its weight of water, after mixing, be warming up to 55 DEG C, add the neutral protease of 500g, enzymolysis 2h under natural pH condition, constantly stir during enzymolysis, enzymolysis terminates, and obtains abalone enzymolysis solution.

[0082] S2, abalone enzymolysis liquid is centrifuged at 12000rpm for 30min, supernatant is taken, filtrate is obtained after coarse filtration with 200 mesh filter cloth, then filtrate is carried out ultrafiltration by the ultrafiltration membrane separation equipment with a molecular weight cut-off being 3000Da to obtain permeate, then permeate is carried out nanofiltration by the nanofiltration membrane separation equipment with a molecular weight cut-off being 500Da to obtain enriched liquid.

[0083] S3. The enriched liquid is passed into a vacuum concentration tank, and concentrated under reduced pressure to 15°Bx at 60°C to obtain a concentrated liquid.

[0084] The concentrated solution was spray-dried at 180°C to make powder, and then packaged to obtain a product containing anti-aging abalone extract.

[0085] Take 50 kg of fresh abalone raw material, remove the shell, take out the meat and viscera, wash once with normal temperature water, blanch with boiling water for 5 minutes, wash once with normal temperature water, drain, dry at 50 ° C for 48 hours, crush with an ultrafine grinder and sieve to obtain abalone powder with a particle size of less than 120 mesh.

[0086] S1, add 15% food-grade ethanol aqueous solution with a volume percentage of 5 times its weight to abalone powder, after mixing, warm up to 55 DEG C, add 500g neutral protease, enzymolysis 2h under natural pH condition, constantly stir during enzymolysis, enzymolysis ends, and obtains abalone enzymolysis solution.

[0087] S2, abalone enzymolysis liquid is centrifuged at 12000rpm for 30min, supernatant is taken, filtrate is obtained after coarse filtration with 200 mesh filter cloth, then filtrate is carried out ultrafiltration by the ultrafiltration membrane separation equipment with a molecular weight cut-off being 3000Da to obtain permeate, then permeate is carried out nanofiltration by the nanofiltration membrane separation equipment with a molecular weight cut-off being 500Da to obtain enriched liquid.

[0088] S3. The enriched liquid is passed into a vacuum concentration tank, and concentrated under reduced pressure to 15°Bx at 60°C to obtain a concentrated liquid.

[0089] The concentrated solution was spray-dried at 180°C to make powder, and then packaged to obtain a product containing anti-aging abalone extract.

[0090] Take 50 kg of fresh abalone raw material, remove the shell, take out the meat and viscera, wash once with normal temperature water, blanch with boiling water for 5 minutes, wash once with normal temperature water, drain, dry at 50 ° C for 48 hours, crush with an ultrafine grinder and sieve to obtain abalone powder with a particle size of less than 120 mesh.

[0091] S1, add 35% food-grade ethanol aqueous solution with a volume percentage of 5 times its weight to abalone powder, after mixing, warm up to 55 DEG C, add 500g neutral protease, enzymolysis 2h under natural pH condition, constantly stir during enzymolysis, enzymolysis ends, and obtains abalone enzymolysis solution.

[0092] S2, abalone enzymolysis liquid is centrifuged under 12000rpm condition with centrifuge, take supernatant, obtain filtrate after coarse filtration with 200 mesh filter cloth, then filtrate is carried out ultrafiltration by the ultrafiltration membrane separation equipment that molecular weight cut-off is 3000Da to obtain permeate, then permeate is carried out nanofiltration by the nanofiltration membrane separation equipment that molecular weight cut-off is 500Da to obtain enriched liquid.

[0093] S3. The enriched liquid is passed into a vacuum concentration tank, and concentrated under reduced pressure to 15°Bx at 60°C to obtain a concentrated liquid.

[0094] The concentrated solution was spray dried at 180°C to make powder, and the product containing anti-aging abalone extract was obtained after packaging. Test Example 2:

[0095] (1) Flux test during separation and purification using ultrafiltration membrane separation equipment:

[0096] The filtrate obtained by coarse filtration after centrifugation was passed into an ultrafiltration membrane separation device with a molecular weight cutoff of 3000Da. The inlet and outlet pressures of the device were controlled to be 2Mpa, and the flux of the permeate within 1 hour was measured.

[0097] Table 1 Changes in membrane flux during ultrafiltration

[0098]

[0099] From the above results, it can be seen that the membrane flux of Comparative Example 1 decreases faster than that of Example 1, the membrane flux changes of Comparative Example 2 and Example 1 are similar, and the membrane flux of Comparative Example 3 decreases rapidly and is obviously seriously blocked. It shows that enzymatic hydrolysis of abalone in an ethanol solution of suitable concentration is helpful to remove impurities, slow down the decline of membrane flux caused by impurities, and thus accelerate the efficiency of ultrafiltration membrane separation and purification.

[0100] (2) Test on DPPH removal effect:

[0101] The DPPH scavenging effect of the products containing the anti-aging abalone extract in Example 1, Comparative Example 1, Comparative Example 2, and Comparative Example 3 was tested using the same testing method as in Test Example 1. The test results are shown in FIG. Figure 2 As shown:

[0102] The above experiment found that at the same concentration, the product obtained in Example 1 had the highest scavenging rate for DPPH·, which was 87.4%, while that in Comparative Example 1 was 53.5%, that in Comparative Example 2 was 57.1%, and that in Comparative Example 3 was 10.7%, indicating that the product obtained by enzymatic hydrolysis of abalone powder with ethanol at an appropriate concentration has a stronger anti-aging ability.

[0103] Embodiment 2:

[0104] Take 50 kg of fresh abalone raw material, remove the shell, take out the meat and viscera, wash once with normal temperature water, blanch with boiling water for 5 minutes, wash once with normal temperature water, drain, dry at 45 ° C for 45 hours, crush with an ultrafine grinder and sieve to obtain abalone powder with a particle size of less than 120 mesh.

[0105] S1, add 20% food-grade ethanol aqueous solution whose volume percentage of 3 times its weight to abalone powder, after mixing, warm up to 50 DEG C, add 250g neutral protease, enzymolysis 2.5h under natural pH condition, constantly stir during enzymolysis, enzymolysis terminates, and obtains abalone enzymolysis solution.

[0106] S2, abalone enzymolysis liquid is centrifuged at 12000rpm for 30min, supernatant is taken, filtrate is obtained after coarse filtration with 150 mesh filter cloths, then filtrate is carried out ultrafiltration by the ultrafiltration membrane separation equipment with a molecular weight cut-off being 3000Da to obtain permeate, then permeate is carried out nanofiltration by the nanofiltration membrane separation equipment with a molecular weight cut-off being 500Da to obtain enriched liquid.

[0107] S3. The enriched liquid is passed into a vacuum concentration tank, and concentrated under reduced pressure to 20°Bx at 60°C to obtain a concentrated liquid.

[0108] The concentrated liquid is spray-dried and powdered at 180° C., and a product containing the anti-aging abalone extract is obtained after packaging.

[0109] When the concentration of the product containing the anti-aging abalone extract obtained in this example is 10 mg / mL, the scavenging rate of DPPH· is 77.5%.

[0110] Embodiment 3:

[0111] Take 50 kg of fresh abalone raw material, remove the shell, take out the meat and viscera, wash once with normal temperature water, blanch with boiling water for 5 minutes, wash once with normal temperature water, drain, dry at 55 ° C for 40 hours, crush with an ultrafine grinder and sieve to obtain abalone powder with a particle size of less than 120 mesh.

[0112] S1, abalone powder is added with 4 times the volume percentage of 30% food-grade ethanol aqueous solution, after mixing, the temperature is raised to 53 ° C, 300g of neutral protease is added, and enzymolysis is carried out under natural pH conditions for 1.5h, and continuous stirring is performed during the enzymolysis. The enzymolysis ends to obtain an abalone enzymolysate.

[0113] S2, abalone enzymolysis liquid is centrifuged at 12000rpm for 30min, supernatant is taken, filtrate is obtained after coarse filtration with 200 mesh filter cloth, then filtrate is carried out ultrafiltration by the ultrafiltration membrane separation equipment with a molecular weight cut-off being 3000Da to obtain permeate, then permeate is carried out nanofiltration by the nanofiltration membrane separation equipment with a molecular weight cut-off being 500Da to obtain enriched liquid.

[0114] S3. The enriched liquid is passed into a vacuum concentration tank, and concentrated under reduced pressure to 20°Bx at 60°C to obtain a concentrated liquid.

[0115] The concentrated liquid is spray-dried and powdered at 180° C., and a product containing the anti-aging abalone extract is obtained after packaging.

[0116] When the concentration of the product containing the anti-aging abalone extract obtained in this example is 10 mg / mL, the scavenging rate of DPPH· is 81.6%.

[0117] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention without departing from the principles and intent of the present invention.

Claims

1. A method for preparing an anti-aging abalone extract, characterized in that: include: S1, abalone powder and low concentration of ethanol are mixed to obtain mixture, and the mixture is hydrolyzed to obtain abalone hydrolyzate; S2, separating and purifying the abalone hydrolysate obtained in S1 to obtain an enriched solution; S3, concentrating the enriched solution obtained in S2 to obtain a concentrated solution.

2. The method for preparing the anti-aging abalone extract according to claim 1, characterized in that: The concentrated solution obtained in step S3 is further spray-dried to obtain abalone extract powder.

3. The method for preparing the anti-aging abalone extract according to claim 1 or 2, characterized in that: The enzymatic hydrolysis in step S1 is performed by a neutral protease that is active in low-concentration ethanol; preferably, the ethanol in step S1 is food-grade ethanol with a volume percentage of 20-30%.

4. The method for preparing the anti-aging abalone extract according to claim 1 or 2, characterized in that: In the step S1, the mass ratio of abalone powder: low concentration ethanol: neutral protease is 1:3-5:0.005-0.

01.

5. The method for preparing the anti-aging abalone extract according to claim 1 or 2, characterized in that: The abalone powder in step S1 is obtained by pre-treating abalone, drying and crushing the edible part of abalone to obtain abalone powder; Preferably, the pretreatment is to shell the live abalone, take out the meat and viscera, and further wash, blanch, cool and drain to obtain the edible part of the abalone; Preferably, the drying temperature is 45-55°C; Preferably, the edible part of the abalone is dried until it can be crushed into powder; more preferably, the drying time is 40-48 hours.

6. The method for preparing the anti-aging abalone extract according to claim 1 or 2, characterized in that: The temperature of enzymolysis in step S1 is 50-55° C.; preferably, the time of enzymolysis in step S2 is 1.5 h-2.5 h.

7. The method for preparing the anti-aging abalone extract according to claim 1 or 2, characterized in that: The step S2 is to centrifuge the abalone hydrolysate to obtain a supernatant, perform coarse filtration on the supernatant to obtain a filtrate, perform ultrafiltration on the filtrate to obtain a permeate, and perform nanofiltration on the permeate to obtain an enriched solution containing active ingredients; Preferably, the centrifugation is performed at 10000-12000 rpm for 25-30 min; Preferably, the coarse filtration is performed using a filter cloth with a mesh size of 150-250; Preferably, the ultrafiltration is to filter the filtrate using an ultrafiltration membrane separation device with a molecular weight cutoff of 3000Da; Preferably, the nanofiltration is to use a nanofiltration membrane separation device with a molecular weight cutoff of 500Da to intercept the permeate.

8. The method for preparing the anti-aging abalone extract according to claim 1 or 2, characterized in that: The concentration of the concentrated solution is 15-20° Bx; preferably, the enriched solution in step S3 is concentrated at 50-60° C. to obtain the concentrated solution.

9. The abalone extract prepared according to the preparation method according to any one of claims 1 to 8 has anti-aging uses.

10. A product containing an anti-aging abalone extract, characterized in that: The abalone extract prepared by the preparation method according to any one of claims 1 to 8 is prepared into one or more of capsules, tablets, granules, oral liquids or beverages.

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