Extraction method and application of mango extract

Abstract

The invention discloses an extraction method of a mango extract, which comprises the following steps: 1) peeling and denucleating mangoes to obtain mango pulp, and pulping the mango pulp into pulp; 2) performing pulsed electric field extraction on the slurry, and centrifuging to obtain a first supernatant and a first precipitate; 3) performing centrifugal separation on the first precipitate enzymatic hydrolysate to obtain a second supernatant and a second precipitate; (4) mixing the first supernate and the second supernate, and performing ultrafiltration to remove protein with the protein molecular weight of 20-40Kda, so as to obtain a mixture; and 5) mixing and homogenizing the mixture and the second precipitate to obtain the mango extract. The mango extract disclosed by the invention is applied to preparation of the anti-alopecia cream, has a very good anti-alopecia effect, is free from allergic phenomena such as rash, red and swollen skin and blister, is safe and reliable, and has relatively high popularization and application values.

Description

Technical Field

[0001] The present invention relates to the technical field of mango processing and application, and more specifically, to a method for extracting mango extract and application thereof. Background Art

[0002] Mango (Mangifera indica L.) is an evergreen fruit tree of the Anacardiaceae family. It is known as the "King of Tropical Fruits". It is loved by people for its attractive aroma, rich taste and rich nutrition. It is one of the important specialty fruits in Guangxi Zhuang Autonomous Region. The main production area is Baise City. It is concentrated on the market from late June to late August, mainly for fresh consumption or processed dried fruits. The processed products are single and low-priced, and are prone to seasonal and regional sluggish sales.

[0003] Therefore, it is urgent to expand the processing and application market of mangoes and improve the economic benefits of the mango industry. Summary of the invention

[0004] An object of the present invention is to solve at least the above problems and to provide at least the advantages which will be described hereinafter.

[0005] The invention provides a method for extracting mango extract, aiming to obtain a large amount of low molecular weight hydrolases, polypeptides, oligosaccharides, monosaccharides and other active structural components in mango, retain its rich vitamins, minerals, active ingredients, etc., and remove proteins with a molecular weight of 20Kda to 40Kda. The extracted mango extract is used to prepare an anti-hair loss cream, which has a good anti-hair loss effect, has no allergic phenomena such as rashes, redness, swelling, blisters, etc. on the skin, is safe and reliable, and has a high value of promotion and application.

[0006] In order to achieve these purposes and other advantages of the present invention, a method for extracting mango extract is provided, which comprises the following steps:

[0007] 1) peeling and core-removing mangoes, and beating the obtained mango pulp into pulp;

[0008] 2) subjecting the slurry to pulse electric field extraction under the conditions of electric field strength of 10-30 kv / cm, flow rate of 30-40 mL / min, pulse frequency of 400-420 Hz, pulse width of 1-2 μs, pulse time of 450-500 μs, and 50-60 MPa, and centrifuging at 1500-2000 r / min for 2-5 min to obtain a first supernatant and a first precipitate;

[0009] 3) mixing the first precipitate with cellulase, chitin endonuclease, α-amylase and amyloglucosidase with deionized water and performing enzymolysis for 24 to 48 hours to obtain an enzymolyzate, adjusting the pH of the enzymolyzate to 8.0 to 8.5 and centrifuging at 1500 to 2000 r / min for 2 to 5 minutes to obtain a second supernatant and a second precipitate; wherein the weight ratio of the first precipitate to the cellulase, chitin endonuclease, α-amylase, amyloglucosidase and deionized water is 40 to 50:1 to 2:1 to 2:0.5:1:1 to 5, and the enzymolysis temperature is 40-50°C;

[0010] 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 5-6, then adding CaCl2, mixing, placing at 2-4°C for 1-2 hours, and removing proteins with a molecular weight of 20Kda to 40Kda by ultrafiltration to obtain a mixture; wherein the weight ratio of the mixture to CaCl2 is 17-20:1;

[0011] 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 40 to 50 MPa for 10 to 20 minutes to obtain a mango extract.

[0012] Preferably, in step 1), before peeling and pitting the mangoes, the process further comprises refrigerating the mangoes to 5-10° C., which is beneficial to improving the quality of the pulp and facilitating peeling and pitting.

[0013] Preferably, in step 2), the pulse electric field also includes ultrasonic waves with a frequency of 30-70 KHz and a power of 300-500 W.

[0014] Preferably, in step 3), the enzymatic activity of the cellulase is at least 2000 U / g, the enzymatic activity of the chitinase is at least 200 U / g, the enzymatic activity of the α-amylase is at least 3000 U / g, and the enzymatic activity of the amyloglucosidase is at least 1000 U / g.

[0015] Preferably, in step 4), proteins with a molecular weight of 20Kda to 40Kda are removed by ultrafiltration, specifically: first, centrifugal filtration is performed through an ultrafiltration tube with a molecular weight cutoff of 40kDa to obtain a first filtrate and a first concentrated solution, and then the first filtrate is centrifugal filtration through an ultrafiltration tube with a molecular weight cutoff of 20kDa to obtain a second filtrate and a second concentrated solution; the second concentrated solution is obtained by ultrafiltration to remove proteins with a molecular weight of 20Kda to 40Kda, and the second filtrate and the first concentrated solution are mixed to obtain the mixture.

[0016] The present invention also provides the application of the mango extract extracted by the above method. The mango extract obtained by the above extraction method is used to prepare an anti-hair loss cream.

[0017] The present invention has at least the following beneficial effects:

[0018] 1. The present invention uses a high-intensity pulsed electric field to rupture and extract the pulp cells of mango pulp, dissolve the rich nutrients of mango, obtain active structural components, separate the precipitate and perform mixed enzymolysis with cellulase, chitin endonuclease, α-amylase and amyloglucosidase to obtain a large amount of low molecular weight hydrolases, polypeptides, oligosaccharides and monosaccharides in mango, which is beneficial to the absorption of active components; centrifugal stratification, ultrafiltration to remove proteins with a molecular weight of 20Kda to 40Kda, retain its rich vitamins, minerals, active components, etc., so that the extracted mango extract is used to prepare an anti-hair loss cream, which has a good anti-hair loss effect and has no allergic phenomena such as rashes, redness, swelling, blisters, etc. on the skin. It is safe and reliable and has a high value for promotion and application.

[0019] 2. The present invention combines low-intensity pulsed electric field with ultrasonic cavitation to rupture and extract the pulp cells of mango pulp, thereby promoting the dissolution of rich nutrients in mango and obtaining more favorable active structural components, further improving the anti-hair loss effect.

[0020] Other advantages, objectives and features of the present invention will be embodied in part through the following description, and in part will be understood by those skilled in the art through study and practice of the present invention. DETAILED DESCRIPTION

[0021] The present invention is further described in detail below in conjunction with experiments, embodiments and comparative examples so that those skilled in the art can implement the invention with reference to the description.

[0022] It should be understood that the terms such as “having”, “including” and “comprising” used herein do not exclude the existence or addition of one or more other elements or combinations thereof.

[0023] It should be noted that the experimental methods described in the following embodiments are conventional methods unless otherwise specified, and the reagents and materials can be obtained from commercial channels unless otherwise specified.

[0024] <Example 1>

[0025] A method for extracting mango extract comprises the following steps:

[0026] 1) Refrigerate the mangoes to 10° C., take out the mangoes, peel and pit them, and blend the mango pulp into a slurry;

[0027] 2) subjecting the slurry to pulse electric field extraction under the conditions of electric field strength of 30 kv / cm, flow rate of 40 mL / min, pulse frequency of 420 Hz, pulse width of 2 μs, pulse time of 500 μs, and 60 MPa, and centrifuging at 2000 r / min for 5 min to obtain a first supernatant and a first precipitate;

[0028] 3) The first precipitate is mixed with cellulase, chitin endonuclease, α-amylase and amyloglucosidase and deionized water for uniform enzymolysis for 48 hours to obtain an enzymolyzate, and the enzymolyzate is adjusted to pH 8.5 by adding sodium hydroxide, and the enzymolyzate is centrifuged at 2000 r / min for 5 minutes to obtain a second supernatant and a second precipitate; wherein the weight ratio of the first precipitate to cellulase (enzyme activity 2000 U / g), chitin endonuclease (enzyme activity 200 U / g), α-amylase (enzyme activity 3000 U / g), amyloglucosidase (enzyme activity 1000 U / g), and deionized water is 50:2:2:0.5:1:5, and the enzymolysis temperature is 50°C; stirring is performed for 10 minutes every 2 hours during the enzymolysis process;

[0029] 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 6, then adding CaCl2, mixing, placing at 4°C for 2 hours, and removing proteins with a molecular weight of 20Kda to 40Kda by ultrafiltration to obtain a mixture; wherein the weight ratio of the mixture to CaCl2 is 20:1;

[0030] 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 50 MPa for 20 min to obtain a mango extract.

[0031] Among them, the mango adopts Tainong No. 1, and the mango is refrigerated to 10℃, specifically, the core temperature of the mango reaches 10℃. According to the actual operation, the mango can be placed in an environment of 10℃ and refrigerated for 10 hours;

[0032] Wherein, in step 4), proteins with a molecular weight of 20Kda to 40Kda are removed by ultrafiltration, specifically: first, centrifugal filtration is performed through an ultrafiltration tube with a molecular weight cutoff of 40kDa to obtain a first filtrate and a first concentrated solution, and then the first filtrate is centrifugal filtration through an ultrafiltration tube with a molecular weight cutoff of 20kDa to obtain a second filtrate and a second concentrated solution; the second concentrated solution is the protein with a molecular weight of 20Kda to 40Kda removed by ultrafiltration, and the second filtrate and the first concentrated solution are mixed to obtain the mixture.

[0033] <Example 2>

[0034] A method for extracting mango extract comprises the following steps:

[0035] 1) Refrigerate the mangoes to 7° C., remove the skin and core, and blend the mango pulp into a slurry;

[0036] 2) subjecting the slurry to pulse electric field extraction under the conditions of electric field strength of 20 kv / cm, flow rate of 35 mL / min, pulse frequency of 410 Hz, pulse width of 1.5 μs, pulse time of 470 μs, and 55 MPa, and centrifuging at 1700 r / min for 3 min to obtain a first supernatant and a first precipitate;

[0037] 3) the first precipitate is mixed with cellulase, chitin endonuclease, α-amylase and amyloglucosidase and deionized water for uniform enzymolysis for 36 hours to obtain an enzymolyzate, sodium hydroxide is added to adjust the pH of the enzymolyzate to 8.0, and the enzymolyzate is centrifuged at 1700r / min for 3min to obtain a second supernatant and a second precipitate; wherein the weight ratio of the first precipitate to cellulase (enzyme activity 2000U / g), chitin endonuclease (enzyme activity 200U / g), α-amylase (enzyme activity 3000U / g), amyloglucosidase (enzyme activity 1000U / g), and deionized water is 45:1.5:1.5:0.5:1:4, and the enzymolysis temperature is 45°C; stirring is performed for 10 minutes every 2 hours during the enzymolysis process;

[0038] 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 5.5, then adding CaCl2, mixing and placing at 3°C ​​for 1.5 hours, removing proteins with a molecular weight of 20Kda to 40Kda by ultrafiltration, and obtaining a mixture; wherein the weight ratio of the mixture to CaCl2 is 18:1;

[0039] 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 45 MPa for 15 min to obtain a mango extract.

[0040] Among them, the mangoes are Tainong No. 1, and the mangoes are refrigerated to 7°C, specifically, the core temperature of the mangoes reaches 7°C. According to actual operation, the mangoes can be placed in a 7°C environment and refrigerated for 8 hours;

[0041] Wherein, in step 4), proteins with a molecular weight of 20Kda to 40Kda are removed by ultrafiltration, specifically: first, centrifugal filtration is performed through an ultrafiltration tube with a molecular weight cutoff of 40kDa to obtain a first filtrate and a first concentrated solution, and then the first filtrate is centrifugal filtration through an ultrafiltration tube with a molecular weight cutoff of 20kDa to obtain a second filtrate and a second concentrated solution; the second concentrated solution is the protein with a molecular weight of 20Kda to 40Kda removed by ultrafiltration, and the second filtrate and the first concentrated solution are mixed to obtain the mixture.

[0042] <Example 3>

[0043] A method for extracting mango extract comprises the following steps:

[0044] 1) Refrigerate the mangoes to 5° C., take out the mangoes, peel and pit them, and blend the mango pulp into a slurry;

[0045] 2) subjecting the slurry to pulse electric field extraction under the conditions of electric field strength of 10 kv / cm, flow rate of 30 mL / min, pulse frequency of 400 Hz, pulse width of 1 μs, pulse time of 450 μs, and 50 MPa, and then centrifuging at 1500 r / min for 2 min to obtain a first supernatant and a first precipitate;

[0046] 3) The first precipitate is mixed with cellulase, chitin endonuclease, α-amylase and amyloglucosidase and deionized water for uniform enzymolysis for 24 hours to obtain an enzymolyzate, and the enzymolyzate is adjusted to pH 8.0 by adding sodium hydroxide, centrifuged at 1500r / min for 2min to obtain a second supernatant and a second precipitate; wherein the weight ratio of the first precipitate to cellulase (enzyme activity 2000U / g), chitin endonuclease (enzyme activity 200U / g), α-amylase (enzyme activity 3000U / g), amyloglucosidase (enzyme activity 1000U / g), and deionized water is 40:1:1:0.5:1:3, and the enzymolysis temperature is 40°C; stirring for 10 minutes every 2 hours during the enzymolysis process;

[0047] 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 5, then adding CaCl2, mixing and placing at 2°C for 1 hour, removing proteins with a molecular weight of 20Kda to 40Kda by ultrafiltration, and obtaining a mixture; wherein the weight ratio of the mixture to CaCl2 is 17:1;

[0048] 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 40 MPa for 10 min to obtain a mango extract.

[0049] Among them, the mangoes are made of Tainong No. 1, and the mangoes are refrigerated to 5℃, specifically, the core temperature of the mangoes reaches 5℃. According to the actual operation, the mangoes can be placed in a 5℃ environment and refrigerated for 6 hours;

[0050] Wherein, in step 4), proteins with a molecular weight of 20Kda to 40Kda are removed by ultrafiltration, specifically: first, centrifugal filtration is performed through an ultrafiltration tube with a molecular weight cutoff of 40kDa to obtain a first filtrate and a first concentrated solution, and then the first filtrate is centrifugal filtration through an ultrafiltration tube with a molecular weight cutoff of 20kDa to obtain a second filtrate and a second concentrated solution; the second concentrated solution is the protein with a molecular weight of 20Kda to 40Kda removed by ultrafiltration, and the second filtrate and the first concentrated solution are mixed to obtain the mixture.

[0051] <Example 4>

[0052] A method for extracting mango extract comprises the following steps:

[0053] 1) Refrigerate the mangoes to 5° C., take out the mangoes, peel and pit them, and blend the mango pulp into a slurry;

[0054] 2) subjecting the slurry to ultrasonic pulse electric field extraction under the conditions of electric field strength of 10 kv / cm, flow rate of 30 mL / min, pulse frequency of 400 Hz, pulse width of 1 μs, pulse time of 450 μs, 50 MPa, ultrasonic frequency of 30 KHz, and ultrasonic power of 300 W, and then centrifuging at 1500 r / min for 2 min to obtain a first supernatant and a first precipitate;

[0055] 3) The first precipitate is mixed with cellulase, chitin endonuclease, α-amylase and amyloglucosidase and deionized water for uniform enzymolysis for 24 hours to obtain an enzymolyzate, and the enzymolyzate is adjusted to pH 8.0 by adding sodium hydroxide, centrifuged at 1500r / min for 2min to obtain a second supernatant and a second precipitate; wherein the weight ratio of the first precipitate to cellulase (enzyme activity 2000U / g), chitin endonuclease (enzyme activity 200U / g), α-amylase (enzyme activity 3000U / g), amyloglucosidase (enzyme activity 1000U / g), and deionized water is 40:1:1:0.5:1:3, and the enzymolysis temperature is 40°C; stirring for 10 minutes every 2 hours during the enzymolysis process;

[0056] 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 5, then adding CaCl2, mixing and placing at 2°C for 1 hour, removing proteins with a molecular weight of 20Kda to 40Kda by ultrafiltration, and obtaining a mixture; wherein the weight ratio of the mixture to CaCl2 is 17:1;

[0057] 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 40 MPa for 10 min to obtain a mango extract.

[0058] Among them, the mangoes are made of Tainong No. 1, and the mangoes are refrigerated to 5℃, specifically, the core temperature of the mangoes reaches 5℃. According to the actual operation, the mangoes can be placed in a 5℃ environment and refrigerated for 6 hours;

[0059] Wherein, in step 4), proteins with a molecular weight of 20Kda to 40Kda are removed by ultrafiltration, specifically: first, centrifugal filtration is performed through an ultrafiltration tube with a molecular weight cutoff of 40kDa to obtain a first filtrate and a first concentrated solution, and then the first filtrate is centrifugal filtration through an ultrafiltration tube with a molecular weight cutoff of 20kDa to obtain a second filtrate and a second concentrated solution; the second concentrated solution is the protein with a molecular weight of 20Kda to 40Kda removed by ultrafiltration, and the second filtrate and the first concentrated solution are mixed to obtain the mixture.

[0060] Comparative Example 1

[0061] On the basis of Example 4, step 2) does not involve pulse electric field extraction, and the rest is the same as Example 4. Specifically:

[0062] A method for extracting mango extract comprises the following steps:

[0063] 1) Refrigerate the mangoes to 5° C., take out the mangoes, peel and pit them, and blend the mango pulp into a slurry;

[0064] 2) subjecting the slurry to ultrasonic extraction at 50 MPa, an ultrasonic frequency of 30 KHz, and an ultrasonic power of 300 W, and then centrifuging at 1500 r / min for 2 min to obtain a first supernatant and a first precipitate;

[0065] 3) The first precipitate is mixed with cellulase, chitin endonuclease, α-amylase and amyloglucosidase and deionized water for uniform enzymolysis for 24 hours to obtain an enzymolyzate, and the enzymolyzate is adjusted to pH 8.0 by adding sodium hydroxide, centrifuged at 1500r / min for 2min to obtain a second supernatant and a second precipitate; wherein the weight ratio of the first precipitate to cellulase (enzyme activity 2000U / g), chitin endonuclease (enzyme activity 200U / g), α-amylase (enzyme activity 3000U / g), amyloglucosidase (enzyme activity 1000U / g), and deionized water is 40:1:1:0.5:1:3, and the enzymolysis temperature is 40°C; stirring for 10 minutes every 2 hours during the enzymolysis process;

[0066] 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 5, then adding CaCl2, mixing and placing at 2°C for 1 hour, removing proteins with a molecular weight of 20Kda to 40Kda by ultrafiltration, and obtaining a mixture; wherein the weight ratio of the mixture to CaCl2 is 17:1;

[0067] 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 40 MPa for 10 min to obtain a mango extract.

[0068] Among them, the mangoes are made of Tainong No. 1, and the mangoes are refrigerated to 5℃, specifically, the core temperature of the mangoes reaches 5℃. According to the actual operation, the mangoes can be placed in a 5℃ environment and refrigerated for 6 hours;

[0069] Wherein, in step 4), proteins with a molecular weight of 20Kda to 40Kda are removed by ultrafiltration, specifically: first, centrifugal filtration is performed through an ultrafiltration tube with a molecular weight cutoff of 40kDa to obtain a first filtrate and a first concentrated solution, and then the first filtrate is centrifugal filtration through an ultrafiltration tube with a molecular weight cutoff of 20kDa to obtain a second filtrate and a second concentrated solution; the second concentrated solution is the protein with a molecular weight of 20Kda to 40Kda removed by ultrafiltration, and the second filtrate and the first concentrated solution are mixed to obtain the mixture.

[0070] Comparative Example 2

[0071] On the basis of Example 4, step 3) does not include enzymolysis, and the rest is the same as Example 4. Specifically:

[0072] A method for extracting mango extract comprises the following steps:

[0073] 1) Refrigerate the mangoes to 5° C., take out the mangoes, peel and pit them, and blend the mango pulp into a slurry;

[0074] 2) subjecting the slurry to ultrasonic pulse electric field extraction under the conditions of electric field strength of 10 kv / cm, flow rate of 30 mL / min, pulse frequency of 400 Hz, pulse width of 1 μs, pulse time of 450 μs, 50 MPa, ultrasonic frequency of 30 KHz, and ultrasonic power of 300 W, and then centrifuging at 1500 r / min for 2 min to obtain a first supernatant and a first precipitate;

[0075] 3) adjusting the pH of the first precipitate to 8.0 by sodium hydroxide, and then centrifuging at 1500 r / min for 2 min to obtain a second supernatant and a second precipitate;

[0076] 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 5, then adding CaCl2, mixing and placing at 2°C for 1 hour, removing proteins with a molecular weight of 20Kda to 40Kda by ultrafiltration, and obtaining a mixture; wherein the weight ratio of the mixture to CaCl2 is 17:1;

[0077] 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 40 MPa for 10 min to obtain a mango extract.

[0078] Among them, the mangoes are made of Tainong No. 1, and the mangoes are refrigerated to 5℃, specifically, the core temperature of the mangoes reaches 5℃. According to the actual operation, the mangoes can be placed in a 5℃ environment and refrigerated for 6 hours;

[0079] Wherein, in step 4), proteins with a molecular weight of 20Kda to 40Kda are removed by ultrafiltration, specifically: first, centrifugal filtration is performed through an ultrafiltration tube with a molecular weight cutoff of 40kDa to obtain a first filtrate and a first concentrated solution, and then the first filtrate is centrifugal filtration through an ultrafiltration tube with a molecular weight cutoff of 20kDa to obtain a second filtrate and a second concentrated solution; the second concentrated solution is the protein with a molecular weight of 20Kda to 40Kda removed by ultrafiltration, and the second filtrate and the first concentrated solution are mixed to obtain the mixture.

[0080] Comparative Example 3

[0081] On the basis of Example 4, step 4) does not include ultrafiltration to remove proteins with a molecular weight of 20Kda to 40Kda, and the rest is the same as Example 4. Specifically:

[0082] A method for extracting mango extract comprises the following steps:

[0083] 1) Refrigerate the mangoes to 5° C., take out the mangoes, peel and pit them, and blend the mango pulp into a slurry;

[0084] 2) subjecting the slurry to ultrasonic pulse electric field extraction under the conditions of electric field strength of 10 kv / cm, flow rate of 30 mL / min, pulse frequency of 400 Hz, pulse width of 1 μs, pulse time of 450 μs, 50 MPa, ultrasonic frequency of 30 KHz, and ultrasonic power of 300 W, and then centrifuging at 1500 r / min for 2 min to obtain a first supernatant and a first precipitate;

[0085] 3) The first precipitate is mixed with cellulase, chitin endonuclease, α-amylase and amyloglucosidase and deionized water for uniform enzymolysis for 24 hours to obtain an enzymolyzate, and the enzymolyzate is adjusted to pH 8.0 by adding sodium hydroxide, centrifuged at 1500r / min for 2min to obtain a second supernatant and a second precipitate; wherein the weight ratio of the first precipitate to cellulase (enzyme activity 2000U / g), chitin endonuclease (enzyme activity 200U / g), α-amylase (enzyme activity 3000U / g), amyloglucosidase (enzyme activity 1000U / g), and deionized water is 40:1:1:0.5:1:3, and the enzymolysis temperature is 40°C; stirring for 10 minutes every 2 hours during the enzymolysis process;

[0086] 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 5, then adding CaCl2, mixing well, placing at 2°C and standing for 1 hour to obtain a mixture; wherein the weight ratio of the mixture to CaCl2 is 17:1;

[0087] 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 40 MPa for 10 min to obtain a mango extract.

[0088] Among them, the mango uses Tainong No. 1, and the mango is refrigerated to 5℃, specifically, the center temperature of the mango reaches 5℃. According to actual operation, the mango can be placed in an environment of 5℃ and refrigerated for 6 hours.

[0089] Comparative Example 4

[0090] On the basis of Example 4, in step 3), chitin endo-nuclease, α-amylase and amyloglucosidase are replaced by chitin exo-nuclease, β-amylase and glucose isomerase respectively, and the rest is the same as Example 4.

[0091] In order to further illustrate the extraction method of the mango extract of the present invention and the technical effects of its application, the present invention conducted experiments on preparing creams with the mango extracts extracted from the above Examples 1-4 and Comparative Examples 1-4.

[0092] 1. Cream preparation

[0093] The creams of Examples 1-4 and Comparative Examples 1-4 are prepared as follows: the mango extracts of Example 1, Example 2, Example 3, Example 4, Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4 are respectively placed in a colloid mill and homogenized for 5 minutes at 40 MPa, wherein the weight ratio of mango extract to monostearate, stearic acid, castor oil, and glycerol is 20:3:1:4:2. The creams of Example 1, Example 2, Example 3, Example 4, Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4 are obtained by homogenization.

[0094] 2. Animal Experiment Materials

[0095] 110 SPF healthy male BALB / C mice were from the same batch of animals with basically the same body shape and age. The experimental animals were kept in a controlled environment with a room temperature of 18-24°C and a humidity of 40%-50%. During the experiment, the animals were free to eat and drink, and their circadian rhythm was normal.

[0096] 3. Main reagent materials and configuration

[0097] Minoxidil ointment (active ingredient content 2%); testosterone propionate injection; soybean oil for injection.

[0098] Preparation method of testosterone propionate solution: dilute testosterone propionate injection and soybean oil for injection at a ratio of 1:5 (V / V) to prepare a testosterone propionate solution with a concentration of 5g / L for use.

[0099] 4. Main instruments and equipment

[0100] 1 / 1000 precision balance.

[0101] 5. Animal Modeling and Drug Administration

[0102] 5.1 Testosterone propionate-induced hair loss model in mice

[0103] 110 male BALB / C mice were adaptively fed for 1 week and randomly divided into 11 groups, 10 mice in each group: blank control group, model control group, minoxidil ointment positive drug group (100 mg / mouse / time), cream group of Example 1 (100 mg / mouse / time), cream group of Example 2 (100 mg / mouse / time), cream group of Example 3 (100 mg / mouse / time), cream group of Example 4 (100 mg / mouse / time), cream group of Comparative Example 1 (100 mg / mouse / time), cream group of Comparative Example 2 (100 mg / mouse / time), cream group of Comparative Example 3 (100 mg / mouse / time), cream group of Comparative Example 4 (100 mg / mouse / time). Before the experiment, testosterone propionate injection and soybean oil for injection were diluted at 1:5 (V / V) to prepare a 5g / L testosterone propionate solution for standby use. Mice in the blank control group were subcutaneously injected with soybean oil for injection on the back of the body once a day; the rest of the modeling groups (model control group, minoxidil liniment positive drug group, Example 1 cream group, Example 2 cream group, Example 3 cream group, Example 4 cream group, Comparative Example 1 cream group, Comparative Example 2 cream group, Comparative Example 3 cream group, Comparative Example 4 cream group) were subcutaneously injected with testosterone propionate solution on the back of the mice for modeling, with a dose of 5 mg / kg, once a day. At the same time, the corresponding drugs were applied to the back hair and hair root skin of each group of mice once a day. The minoxidil ointment positive drug group was given minoxidil ointment 100 mg / mouse / time; the cream group of Example 1, the cream group of Example 2, the cream group of Example 3, the cream group of Example 4, the cream group of Comparative Example 1, the cream group of Comparative Example 2, the cream group of Comparative Example 3, and the cream group of Comparative Example 4 were respectively given the corresponding cream 100 mg / mouse / time; the blank control group and the model control group were given blank matrix 100 mg / mouse / time, and the injection of testosterone propionate solution and the application were continued for 10 weeks.

[0104] 5.2 Detection indicators

[0105] (1) Observe the normal condition of each group of mice and the skin condition of the application site;

[0106] (2) From the fifth week to the end of the experiment, the shed hair of each group of mice was collected every day, and the hair on the back of each mouse was combed at a fixed time period. The shed hair was collected with tweezers and weighed to investigate the degree of hair loss in the mice;

[0107] 6. Experimental Results

[0108] 6.1 Anti-hair loss results and skin conditions in mice

[0109] Model control group: After 2 weeks, the smoothness of the mice's hair decreased; after 4-5 weeks, the hair on the back of the mice gradually became dry and lost its luster; after 7 weeks, the hair on the back of the mice gradually fell off.

[0110] Cream groups of Examples 1-4: All of them can keep hair smooth and improve its glossiness, and significantly improve the hair loss of mice, especially the cream group of Example 4, which has the most significant effect.

[0111] Comparative Example 1-4 cream group: The hair smoothness and glossiness were improved compared with the model control group, but the anti-hair loss effect was worse than that of the Example 1-4 cream group, and skin rashes, redness, swelling, blisters and other phenomena were more common, especially the comparative example 3 cream group had the most obvious skin allergy. The hair loss weight and skin condition of each group of mice are shown in Table 1.

[0112] Table 1

[0113]

[0114]

[0115] It can be seen from the experimental data in Table 1 that in Example 1 and Example 2, the pulp cells of mango pulp are broken by high-intensity pulsed electric field, the rich nutrients of mango are dissolved, the active structural components are obtained, and the separated precipitate is mixed with cellulase, chitin endonuclease, α-amylase and amyloglucosidase for enzymatic hydrolysis to obtain a large amount of low molecular weight hydrolases, polypeptides, oligosaccharides and monosaccharides in mango, which is conducive to the absorption of active ingredients; centrifugal stratification, ultrafiltration to remove proteins with a molecular weight of 20Kda to 40Kda, retaining its rich vitamins, minerals, active ingredients, etc., so that the extracted mango extract is used to prepare anti-hair loss cream, which has a good anti-hair loss effect, and has no allergic phenomena such as rash, redness, and blisters on the skin, which is safe and reliable, and has a high value for promotion and application. Example 3 extracts the pulp cells of mango pulp by low-intensity pulsed electric field, affects the acquisition of active structural components, affects the components of mango extract, and the anti-hair loss effect and skin condition are worse than those of Example 1 and Example 2. Example 4 introduces ultrasonic cavitation on the basis of the low-intensity pulsed electric field in Example 3, which promotes the dissolution of rich nutrients in mango and obtains more favorable active structural components, and can reduce the loss of vitamins, minerals, active structural components and other substances. It not only obviously overcomes the adverse effects of the low-intensity pulsed electric field in Example 3, but also significantly improves the anti-hair loss effect.

[0116] Comparative Example 1 does not have the penetration and oscillating potential of the pulsed electric field, which affects the extraction of the active ingredient structure and the composition of the mango extract, and the anti-hair loss effect is significantly worse than that of Example 4. Comparative Example 2 does not enzymatically hydrolyze the separated precipitate, which affects the absorption and utilization of the active ingredient and the anti-hair loss effect. The product component structure obtained by enzymatic hydrolysis of different composite enzyme combinations in Comparative Example 4 is different, which affects the anti-hair loss effect. Comparative Example 3 failed to remove the adverse protein components with a protein molecular weight of 20Kda to 40Kda, causing serious allergic phenomena such as skin rashes, redness, swelling, and blisters, severe restlessness and fighting, and increased weight of hair loss.

[0117] The number of devices and processing scales described here are used to simplify the description of the present invention. Applications, modifications and variations of the present invention will be obvious to those skilled in the art.

[0118] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes, and they can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to the specific details and the examples shown and described herein.

Claims

1. A method for extracting mango extract, characterized in that: The following steps are involved: 1) peeling and core-removing mangoes, and beating the obtained mango pulp into pulp; 2) subjecting the slurry to pulse electric field extraction under the conditions of electric field strength of 10-30 kv / cm, flow rate of 30-40 mL / min, pulse frequency of 400-420 Hz, pulse width of 1-2 μs, pulse time of 450-500 μs, and 50-60 MPa, and centrifuging at 1500-2000 r / min for 2-5 min to obtain a first supernatant and a first precipitate; 3) mixing the first precipitate with cellulase, chitin endonuclease, α-amylase and amyloglucosidase with deionized water and performing enzymolysis for 24 to 48 hours to obtain an enzymolyzate, adjusting the pH of the enzymolyzate to 8.0 to 8.5 and centrifuging at 1500 to 2000 r / min for 2 to 5 minutes to obtain a second supernatant and a second precipitate; wherein the weight ratio of the first precipitate to the cellulase, chitin endonuclease, α-amylase, amyloglucosidase and deionized water is 40 to 50:1 to 2:1 to 2:0.5:1:1 to 5, and the enzymolysis temperature is 40-50°C; 4) adding hydrochloric acid to the mixture of the first supernatant and the second supernatant to adjust the pH to 5-6, then adding CaCl2, mixing, placing at 2-4°C for 1-2 hours, and removing proteins with a molecular weight of 20Kda to 40Kda by ultrafiltration to obtain a mixture; wherein the weight ratio of the mixture to CaCl2 is 17-20:1; 5) mixing the mixture with the second precipitate, adjusting the pH to neutral, and homogenizing with a colloid mill at 40 to 50 MPa for 10 to 20 minutes to obtain a mango extract.

2. The method for extracting mango extract according to claim 1, characterized in that: In step 1), before peeling and core removal of mangoes, the mangoes are refrigerated at 5-10°C.

3. The method for extracting mango extract according to claim 1, characterized in that: In step 2), the pulse electric field also includes ultrasonic waves with a frequency of 30-70 KHz and an ultrasonic power of 300-500 W.

4. The method for extracting mango extract according to claim 1, characterized in that: In step 3), the enzymatic activity of the cellulase is at least 2000 U / g, the enzymatic activity of the chitinase is at least 200 U / g, the enzymatic activity of the α-amylase is at least 3000 U / g, and the enzymatic activity of the amyloglucosidase is at least 1000 U / g.

5. The method for extracting mango extract according to claim 1, characterized in that: In step 4), proteins with a molecular weight of 20Kda to 40Kda are removed by ultrafiltration, specifically: first, centrifugal filtration is performed through an ultrafiltration tube with a molecular weight cutoff of 40kDa to obtain a first filtrate and a first concentrated solution, and then the first filtrate is centrifugal filtration through an ultrafiltration tube with a molecular weight cutoff of 20kDa to obtain a second filtrate and a second concentrated solution; the second concentrated solution is obtained by ultrafiltration to remove proteins with a molecular weight of 20Kda to 40Kda, and the second filtrate and the first concentrated solution are mixed to obtain the mixture.

6. The use of mango extract, characterized in that: The mango extract obtained by the extraction method according to any one of claims 1 to 5 is used to prepare an anti-hair loss cream.

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