Anti-diabetic potato peptide nutritional supplement and preparation method thereof

Abstract

The invention provides an anti-diabetic potato peptide nutritional supplement and a preparation method thereof, and belongs to the technical field of functional foods. The preparation process comprises the following steps: preparing the potato peptide; preparing amino functionalized mesoporous silica; preparing modified mesoporous silica; preparing potato peptide loaded nanoparticles; preparing microspheres loaded with the mango leaf extract; and preparing the potato peptide nutritional supplement. The preparation method comprises the following steps: activating a taurocholic acid solution, mixing the taurocholic acid solution with an ethylene glycol chitosan solution, carrying out a coupling reaction to prepare a modifier, carrying out modification on amino-functionalized mesoporous silica by using the modifier, loading potato peptide by using the modified mesoporous silica, and preparing the modified potato peptide. The potato peptide can be protected from being damaged by an acid environment and protease in gastric juice, so that the bioavailability of the potato peptide is effectively improved.

Description

Technical Field

[0001] The invention relates to the technical field of functional foods, and in particular to an anti-diabetic potato peptide nutritional supplement and a preparation method thereof. Background Art

[0002] Potato peptides are made from wastewater from potato starch production through microbial fermentation and can be used as food, health products, and pharmaceutical additives. Potato peptides have higher biological activity and nutritional value than the same amount of amino acids. Potato peptides can be completely absorbed and utilized by epithelial cells, enter the blood, and then transported to various parts of the human body to realize their utilization value. Therefore, potato peptides are absorbed by the human body faster than the same amount of free amino acids and much faster than the same amount of protein. Therefore, applying potato peptides to food, health products, and pharmaceuticals can improve effects and reduce costs.

[0003] However, during the oral administration of potato peptides, the complex pH environment in the gastrointestinal tract and the presence of multiple proteases can cause the peptide bonds of potato peptides to break, causing changes in their structure and function, resulting in a loss of efficacy, which in turn leads to low bioavailability of potato peptides.

[0004] Therefore, it is necessary to provide an anti-diabetic potato peptide nutritional supplement with good stability and a preparation method thereof to improve the bioavailability of potato peptides. Summary of the invention

[0005] In view of the deficiencies in the prior art, the object of the present invention is to provide an anti-diabetic potato peptide nutritional supplement and a preparation method thereof.

[0006] A method for preparing an anti-diabetic potato peptide nutritional supplement comprises the following steps:

[0007] S1: Preparation of potato peptides

[0008] The waste water from potato starch production is pretreated to obtain pretreated waste, which is then dispersed in distilled water, enzymatically hydrolyzed, and then fermented to obtain potato peptides;

[0009] S2: Preparation of amino-functionalized mesoporous silica

[0010] Mesoporous silica nanoparticles are added to toluene at a solid-liquid ratio of 1 g: (30-40) mL, and ultrasonically dispersed for 20-30 min to obtain a mesoporous silica nanoparticle suspension, and then 3-aminopropyltriethoxysilane is added to the mesoporous silica nanoparticle suspension at a volume ratio of 1: (140-150), heated at 100-120° C., stirred and refluxed for reaction for 10-12 h, cooled, centrifuged, washed and dried to obtain amino-functionalized mesoporous silica;

[0011] S3: Preparation of modified mesoporous silica

[0012] Ethylene glycol chitosan and taurocholic acid are respectively prepared into an ethylene glycol chitosan solution and a taurocholic acid solution, then the taurocholic acid solution is activated, and then the activated taurocholic acid solution is added into the ethylene glycol chitosan solution for reaction to prepare a modifier, and finally, the amino-functionalized mesoporous silica is modified by the modifier to obtain modified mesoporous silica;

[0013] S4: Preparation of potato peptide-loaded nanoparticles

[0014] The potato peptide and the modified mesoporous silica are prepared into a potato peptide solution and a modified mesoporous silica suspension, and then the potato peptide solution is added into the modified mesoporous silica suspension for ultrasonic loading to obtain potato peptide-loaded nanoparticles;

[0015] S5: Preparation of mango leaf extract loaded microspheres

[0016] Lemongrass essential oil, coconut oil, mango leaf extract and Tween 80 are mixed to prepare a mixed oil liquid, then a carboxymethyl chitosan aqueous solution is mixed with the mixed oil liquid, natural vanillin is added to react, anhydrous ethanol is added, stirred to break the emulsion, centrifuged, washed and dried to obtain mango leaf extract loaded microspheres;

[0017] S6: Preparation of potato peptide nutritional supplements

[0018] The potato peptide-loaded nanoparticles and the mango leaf extract-loaded microspheres are fully and uniformly mixed at a mass ratio of (13-15):1 to obtain a potato peptide nutritional supplement.

[0019] Furthermore, S1 specifically includes the following steps:

[0020] S1.1: placing the waste residue and wastewater from potato starch production in a centrifuge for centrifugal separation, removing the wastewater, and then washing the waste residue to obtain pre-treated waste residue;

[0021] S1.2: Add distilled water to the above pre-treated waste residue at a solid-liquid ratio of 1g: (8-10)mL, stir and disperse thoroughly, heat to 35-40°C and keep warm, then add composite protease, keep warm for 1-2h, inactivate the enzyme, and obtain the enzymatic pre-treatment stock solution;

[0022] S1.3: Inoculate the mixed strains into the above enzymatic pretreatment stock solution, ferment at 32-36°C for 16-18h, sterilize at high temperature, centrifuge, concentrate by membrane filtration and freeze-dry to obtain potato peptides.

[0023] Furthermore, S3 specifically includes the following steps:

[0024] S3.1: dissolving ethylene glycol chitosan and taurocholic acid in dimethyl sulfoxide at a solid-liquid ratio of 1 g:(40-50) mL, respectively, to obtain ethylene glycol chitosan solution and taurocholic acid solution;

[0025] S3.2: Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide to the above taurocholic acid solution at a solid-liquid ratio of 1 g:(1-2) g:(20-30) mL, and stir at a rate of 400-500 r / min for 2-3 h to activate and obtain an activated taurocholic acid solution;

[0026] S3.3: adding an equal volume of the above ethylene glycol chitosan solution to the above activated taurocholic acid solution, stirring and reacting for 6-8 hours, dialyzing and freeze-drying to obtain a modifier;

[0027] S3.4: The amino-functionalized mesoporous silica obtained in step S2 is uniformly dispersed in PBS buffer at a solid-liquid ratio of 1g:(35-40)mL, and then the above-mentioned modifier is added, stirred to react for 10-12h, and modified mesoporous silica is obtained by centrifugation, washing and drying, wherein the mass ratio of the modifier to the amino-functionalized mesoporous silica is 1:(115-125).

[0028] Furthermore, S4 specifically includes the following steps:

[0029] S4.1: dissolving the potato peptide prepared in step S1.3 in PBS buffer at a solid-liquid ratio of 1 g:(25-35) mL to obtain a potato peptide solution;

[0030] S4.2: Add the modified mesoporous silica to PBS buffer at a solid-liquid ratio of 1 g: (45-55) mL, and disperse by ultrasonic for 10-20 min to obtain a modified mesoporous silica suspension;

[0031] S4.3: Add the potato peptide solution to the modified mesoporous silica suspension in a volume ratio of (6-8): 1, stir for 10-12 hours, and then ultrasonically load for 20-30 minutes. After centrifugal separation, washing and drying, potato peptide-loaded nanoparticles are obtained.

[0032] Furthermore, S5 specifically includes the following steps:

[0033] S5.1: lemongrass essential oil and coconut oil are mixed in a volume ratio of 1:(26-30), and then mango leaf extract and Tween 80 are added, heated and stirred at 40-50° C. for 20-30 min, and mixed thoroughly to obtain a mixed oil;

[0034] S5.2: adding 1-2 wt % carboxymethyl chitosan aqueous solution to an equal volume of the above mixed oil, stirring for 10-20 min, adding natural vanillin, stirring and reacting for 2-3 h to obtain a mixed emulsion;

[0035] S5.3: Anhydrous ethanol was added to the mixed emulsion in a volume ratio of (3-4): 1, and the mixed emulsion was stirred and demulsified at a rate of 1500-1600 r / min in an ice water bath at 0°C for 20-30 min, and then centrifuged to retain the gel layer. After washing and drying, microspheres loaded with mango leaf extract were obtained.

[0036] Furthermore, the composite protease is prepared by mixing trypsin and pepsin in a mass ratio of (1-3):1, and the added amount of the composite protease is 3-5% of the mass of the pre-treated waste residue.

[0037] Furthermore, the mixed bacteria include lactic acid bacteria and yeast, and the added amount of the mixed bacteria is 10-12% of the enzymatic pretreatment stock solution.

[0038] Furthermore, the solid-liquid ratio of mango leaf extract to coconut oil is 1 g: (60-70) mL, and the volume ratio of Tween 80 to coconut oil is 1: (20-30).

[0039] Furthermore, the solid-to-liquid ratio of natural vanillin to carboxymethyl chitosan aqueous solution is 1 g: (25-30) mL.

[0040] An anti-diabetic potato peptide nutritional supplement is prepared by any of the above-mentioned methods for preparing an anti-diabetic potato peptide nutritional supplement.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] 1. The present invention first activates a taurocholic acid solution by using 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide, and then mixes the taurocholic acid solution with an ethylene glycol chitosan solution for coupling reaction to prepare a modifier, and then uses the modifier to modify amino-functionalized mesoporous silica. After the modified mesoporous silica is used to load potato peptides, on the one hand, the modifier can provide more active sites, thereby increasing the loading rate of the mesoporous silica for the potato peptides. On the other hand, due to the loading effect of the modified mesoporous silica, the potato peptides can be protected from the acidic environment and protease in the gastric juice, thereby effectively improving the bioavailability of the potato peptides.

[0043] 2. The present invention prepares a mixed oil by first dissolving the mango leaf extract in a mixture of lemongrass essential oil and coconut oil, then fully mixing the mixed oil with a carboxymethyl chitosan aqueous solution, and then adding natural vanillin for cross-linking reaction to form a hydrogel to prepare gel microspheres loaded with the mango leaf extract. Since the carboxyl groups in the carboxymethyl chitosan molecules are deprotonated in an alkaline medium, the molecular chains are extended, and protonated under acidic conditions, the molecular chains shrink to maintain the stability of the gel, thereby protecting the mango leaf extract from being degraded by gastric juice, maintaining the activity of the mango leaf extract, and further improving its bioavailability.

[0044] 3. When the potato peptide-loaded nanoparticles and mango leaf extract-loaded microspheres are mixed to prepare the potato peptide nutritional supplement, since the potato peptides and mango leaf extracts can inhibit the activity of α-glucosidase through different mechanisms, and the potato peptides can improve insulin sensitivity, while the mango leaf extract can promote insulin secretion, the combination of the two can interact with each other and synergistically promote the anti-diabetic effect of the nutritional supplement to more comprehensively regulate blood sugar levels. DETAILED DESCRIPTION

[0045] The following is a detailed description of an anti-diabetic potato peptide nutritional supplement and a preparation method thereof provided by the present invention in conjunction with specific embodiments. At the same time, it is explained here that in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art may also adopt other alternative methods to implement some known technologies.

[0046] Example 1

[0047] A method for preparing an anti-diabetic potato peptide nutritional supplement comprises the following steps:

[0048] S1: Preparation of potato peptides

[0049] S1.1: placing the waste residue and wastewater from potato starch production in a centrifuge for centrifugal separation, removing the wastewater, and then washing the waste residue to obtain pre-treated waste residue;

[0050] S1.2: Add distilled water to the above-mentioned pre-treated waste residue at a solid-liquid ratio of 1g:8mL, stir and disperse thoroughly, heat to 35°C and keep warm, then add composite protease, keep warm for 1h, and after inactivating the enzyme, obtain enzymatic pre-treatment stock solution, wherein the composite protease is a mixture of trypsin and pepsin at a mass ratio of 1:1, and the amount of composite protease added is 3% of the mass of the pre-treated waste residue;

[0051] S1.3: inoculating the mixed strains into the above enzymatic pretreatment stock solution, fermenting at 32°C for 16 hours, sterilizing at high temperature, centrifuging, concentrating by membrane filtration and freeze drying to obtain potato peptides, wherein the mixed strains include lactic acid bacteria and yeast, and the amount of the mixed strains added is 10% of the enzymatic pretreatment stock solution;

[0052] S2: Preparation of amino-functionalized mesoporous silica

[0053] Mesoporous silica nanoparticles were added to toluene at a solid-liquid ratio of 1 g:30 mL, and ultrasonically dispersed for 20 min to obtain a mesoporous silica nanoparticle suspension. 3-aminopropyltriethoxysilane was then added to the mesoporous silica nanoparticle suspension at a volume ratio of 1:140, and the mixture was heated at 100° C. and stirred under reflux for 10 h. After cooling, the mixture was centrifuged, washed, and dried to obtain amino-functionalized mesoporous silica.

[0054] S3: Preparation of modified mesoporous silica

[0055] S3.1: dissolving ethylene glycol chitosan and taurocholic acid in dimethyl sulfoxide at a solid-liquid ratio of 1 g:40 mL, respectively, to obtain ethylene glycol chitosan solution and taurocholic acid solution;

[0056] S3.2: Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide to the above taurocholic acid solution at a solid-liquid ratio of 1g:1g:20mL, and stir at a rate of 400r / min for 2h to activate, thereby obtaining an activated taurocholic acid solution;

[0057] S3.3: adding an equal volume of the above ethylene glycol chitosan solution to the above activated taurocholic acid solution, stirring and reacting for 6-8 hours, dialyzing and freeze-drying to obtain a modifier;

[0058] S3.4: The amino-functionalized mesoporous silica prepared in step S2 is uniformly dispersed in PBS buffer at a solid-liquid ratio of 1 g:35 mL, and then the above-mentioned modifier is added, stirred for reaction for 10 hours, and then centrifuged, washed and dried to obtain modified mesoporous silica, wherein the mass ratio of the modifier to the amino-functionalized mesoporous silica is 1:115;

[0059] S4: Preparation of potato peptide-loaded nanoparticles

[0060] S4.1: dissolving the potato peptide prepared in step S1.3 in PBS buffer at a solid-liquid ratio of 1 g:25 mL to obtain a potato peptide solution;

[0061] S4.2: Add the modified mesoporous silica into PBS buffer at a solid-liquid ratio of 1 g:45 mL, and disperse by ultrasonic for 10 min to obtain a modified mesoporous silica suspension;

[0062] S4.3: adding the potato peptide solution to the modified mesoporous silica suspension at a volume ratio of 6:1, stirring for 10 h, and then ultrasonically loading for 20 min, centrifugally separating, washing and drying to obtain potato peptide-loaded nanoparticles;

[0063] S5: Preparation of mango leaf extract loaded microspheres

[0064] S5.1: lemongrass essential oil and coconut oil are mixed in a volume ratio of 1:26, and then mango leaf extract and Tween 80 are added, heated and stirred at 40° C. for 20 min, and fully mixed to obtain a mixed oil liquid, wherein the solid-liquid ratio of mango leaf extract to coconut oil is 1 g:60 mL, and the volume ratio of Tween 80 to coconut oil is 1:20;

[0065] S5.2: Add 1 wt% carboxymethyl chitosan aqueous solution to an equal volume of the above mixed oil, stir for 10 min, add natural vanillin, stir and react for 2 h to obtain a mixed emulsion, wherein the solid-liquid ratio of natural vanillin to carboxymethyl chitosan aqueous solution is 1 g:25 mL;

[0066] S5.3: adding anhydrous ethanol to the mixed emulsion at a volume ratio of 3:1, and stirring and demulsifying the mixture at a rate of 1500 r / min for 20 min in a 0°C ice water bath, and then centrifuging to retain the gel layer, and obtaining mango leaf extract-loaded microspheres after washing and drying;

[0067] S6: Preparation of potato peptide nutritional supplements

[0068] The potato peptide-loaded nanoparticles and the mango leaf extract-loaded microspheres were fully and evenly mixed at a mass ratio of 13:1 to obtain a potato peptide nutritional supplement.

[0069] Example 2

[0070] A method for preparing an anti-diabetic potato peptide nutritional supplement comprises the following steps:

[0071] S1: Preparation of potato peptides

[0072] S1.1: placing the waste residue and wastewater from potato starch production in a centrifuge for centrifugal separation, removing the wastewater, and then washing the waste residue to obtain pre-treated waste residue;

[0073] S1.2: Add distilled water to the above-mentioned pre-treated waste residue at a solid-liquid ratio of 1g:9mL, stir and disperse thoroughly, heat to 38°C and keep warm, then add composite protease, keep warm for 1.5h, and after inactivating the enzyme, obtain enzymatic pre-treatment stock solution, wherein the composite protease is a mixture of trypsin and pepsin at a mass ratio of 2:1, and the amount of composite protease added is 4% of the mass of the pre-treated waste residue;

[0074] S1.3: inoculating the mixed strains into the above enzymatic pretreatment stock solution, fermenting at 34°C for 17 hours, sterilizing at high temperature, centrifuging, concentrating by membrane filtration and freeze-drying to obtain potato peptides, wherein the mixed strains include lactic acid bacteria and yeast, and the amount of the mixed strains added is 11% of the enzymatic pretreatment stock solution;

[0075] S2: Preparation of amino-functionalized mesoporous silica

[0076] Mesoporous silica nanoparticles were added to toluene at a solid-liquid ratio of 1 g:35 mL, and ultrasonically dispersed for 25 min to obtain a mesoporous silica nanoparticle suspension. 3-aminopropyltriethoxysilane was then added to the mesoporous silica nanoparticle suspension at a volume ratio of 1:145, and heated and stirred at 110° C. for reflux reaction for 11 h. After cooling, the mixture was centrifuged, washed and dried to obtain amino-functionalized mesoporous silica.

[0077] S3: Preparation of modified mesoporous silica

[0078] S3.1: dissolving ethylene glycol chitosan and taurocholic acid in dimethyl sulfoxide at a solid-liquid ratio of 1 g:45 mL, respectively, to obtain ethylene glycol chitosan solution and taurocholic acid solution;

[0079] S3.2: Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide to the above taurocholic acid solution at a solid-liquid ratio of 1 g:1.5 g:25 mL, and stir at a rate of 450 r / min for 2.5 h to activate and obtain an activated taurocholic acid solution;

[0080] S3.3: adding an equal volume of the above-mentioned ethylene glycol chitosan solution to the above-mentioned activated taurocholic acid solution, stirring and reacting for 7 hours, dialyzing and freeze-drying to obtain a modifier;

[0081] S3.4: The amino-functionalized mesoporous silica prepared in step S2 is uniformly dispersed in PBS buffer at a solid-liquid ratio of 1 g:38 mL, and the above-mentioned modifier is added, stirred for reaction for 11 hours, and then centrifuged, washed and dried to obtain modified mesoporous silica, wherein the mass ratio of the modifier to the amino-functionalized mesoporous silica is 1:120;

[0082] S4: Preparation of potato peptide-loaded nanoparticles

[0083] S4.1: dissolving the potato peptide prepared in step S1.3 in PBS buffer at a solid-liquid ratio of 1 g:30 mL to obtain a potato peptide solution;

[0084] S4.2: Add the modified mesoporous silica into PBS buffer at a solid-liquid ratio of 1 g:50 mL, and disperse by ultrasonic for 15 min to obtain a modified mesoporous silica suspension;

[0085] S4.3: adding the potato peptide solution to the modified mesoporous silica suspension at a volume ratio of 7:1, stirring for 11 hours, and then ultrasonically loading for 25 minutes, centrifugally separating, washing and drying to obtain potato peptide-loaded nanoparticles;

[0086] S5: Preparation of mango leaf extract loaded microspheres

[0087] S5.1: lemongrass essential oil and coconut oil are mixed in a volume ratio of 1:28, and then mango leaf extract and Tween 80 are added, heated and stirred at 45° C. for 25 minutes, and fully mixed to obtain a mixed oil liquid, wherein the solid-liquid ratio of mango leaf extract to coconut oil is 1 g:65 mL, and the volume ratio of Tween 80 to coconut oil is 1:25;

[0088] S5.2: adding 1.5 wt% carboxymethyl chitosan aqueous solution to an equal volume of the above mixed oil, stirring for 15 min, adding natural vanillin, stirring and reacting for 2.5 h, to obtain a mixed emulsion, wherein the solid-liquid ratio of natural vanillin to carboxymethyl chitosan aqueous solution is 1 g:28 mL;

[0089] S5.3: adding anhydrous ethanol to the mixed emulsion at a volume ratio of 3.5:1, and stirring and demulsifying the mixture at a rate of 1550 r / min for 25 min in a 0°C ice-water bath, and then centrifuging the mixture to retain the gel layer, and obtaining mango leaf extract-loaded microspheres after washing and drying;

[0090] S6: Preparation of potato peptide nutritional supplements

[0091] The potato peptide-loaded nanoparticles and the mango leaf extract-loaded microspheres were fully and evenly mixed at a mass ratio of 14:1 to obtain a potato peptide nutritional supplement.

[0092] Example 3

[0093] A method for preparing an anti-diabetic potato peptide nutritional supplement comprises the following steps:

[0094] S1: Preparation of potato peptides

[0095] S1.1: placing the waste residue and wastewater from potato starch production in a centrifuge for centrifugal separation, removing the wastewater, and then washing the waste residue to obtain pre-treated waste residue;

[0096] S1.2: Add distilled water to the above-mentioned pre-treated waste residue at a solid-liquid ratio of 1g:10mL, stir and disperse thoroughly, heat to 40°C and keep warm, then add composite protease, keep warm for 2h, and after inactivating the enzyme, obtain enzymatic pre-treatment stock solution, wherein the composite protease is a mixture of trypsin and pepsin at a mass ratio of 3:1, and the amount of composite protease added is 5% of the mass of the pre-treated waste residue;

[0097] S1.3: inoculating the mixed strains into the above enzymatic pretreatment stock solution, fermenting at 36°C for 18 hours, sterilizing at high temperature, centrifuging, concentrating by membrane filtration and freeze-drying to obtain potato peptides, wherein the mixed strains include lactic acid bacteria and yeast, and the amount of the mixed strains added is 12% of the enzymatic pretreatment stock solution;

[0098] S2: Preparation of amino-functionalized mesoporous silica

[0099] Mesoporous silica nanoparticles were added to toluene at a solid-liquid ratio of 1 g:40 mL, and ultrasonically dispersed for 30 min to obtain a mesoporous silica nanoparticle suspension. 3-aminopropyltriethoxysilane was then added to the mesoporous silica nanoparticle suspension at a volume ratio of 1:150, and heated and stirred at 120° C. for reflux reaction for 12 h. After cooling, the mixture was centrifuged, washed and dried to obtain amino-functionalized mesoporous silica.

[0100] S3: Preparation of modified mesoporous silica

[0101] S3.1: dissolving ethylene glycol chitosan and taurocholic acid in dimethyl sulfoxide at a solid-liquid ratio of 1 g:50 mL, respectively, to obtain ethylene glycol chitosan solution and taurocholic acid solution;

[0102] S3.2: Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide to the above taurocholic acid solution at a solid-liquid ratio of 1g:2g:30mL, and stir at a rate of 500r / min for 3h to activate, thereby obtaining an activated taurocholic acid solution;

[0103] S3.3: adding an equal volume of the above ethylene glycol chitosan solution to the above activated taurocholic acid solution, stirring and reacting for 8 hours, dialyzing and freeze-drying to obtain a modifier;

[0104] S3.4: The amino-functionalized mesoporous silica prepared in step S2 is uniformly dispersed in PBS buffer at a solid-liquid ratio of 1 g:40 mL, and then the above-mentioned modifier is added, stirred for reaction for 12 hours, and then centrifuged, washed and dried to obtain modified mesoporous silica, wherein the mass ratio of the modifier to the amino-functionalized mesoporous silica is 1:125;

[0105] S4: Preparation of potato peptide-loaded nanoparticles

[0106] S4.1: dissolving the potato peptide prepared in step S1.3 in PBS buffer at a solid-liquid ratio of 1 g:35 mL to obtain a potato peptide solution;

[0107] S4.2: Add the modified mesoporous silica into PBS buffer at a solid-liquid ratio of 1 g:55 mL, and disperse by ultrasonic for 20 min to obtain a modified mesoporous silica suspension;

[0108] S4.3: adding the potato peptide solution to the modified mesoporous silica suspension at a volume ratio of 8:1, stirring for 12 hours, and then ultrasonically loading for 30 minutes, centrifugally separating, washing and drying to obtain potato peptide-loaded nanoparticles;

[0109] S5: Preparation of mango leaf extract loaded microspheres

[0110] S5.1: lemongrass essential oil and coconut oil are mixed in a volume ratio of 1:30, and then mango leaf extract and Tween 80 are added, heated and stirred at 50° C. for 30 min, and fully mixed to obtain a mixed oil liquid, wherein the solid-liquid ratio of mango leaf extract to coconut oil is 1 g:70 mL, and the volume ratio of Tween 80 to coconut oil is 1:30;

[0111] S5.2: adding 2 wt% carboxymethyl chitosan aqueous solution to an equal volume of the above mixed oil, stirring for 20 min, adding natural vanillin, stirring and reacting for 3 h to obtain a mixed emulsion, wherein the solid-liquid ratio of natural vanillin to carboxymethyl chitosan aqueous solution is 1 g:30 mL;

[0112] S5.3: adding anhydrous ethanol to the mixed emulsion at a volume ratio of 4:1, and stirring and demulsifying the mixture at a rate of 1600 r / min for 30 min in a 0°C ice water bath, and then centrifuging to retain the gel layer, and obtaining mango leaf extract-loaded microspheres after washing and drying;

[0113] S6: Preparation of potato peptide nutritional supplements

[0114] The potato peptide-loaded nanoparticles and the mango leaf extract-loaded microspheres were fully and uniformly mixed at a mass ratio of 15:1 to obtain a potato peptide nutritional supplement.

[0115] Comparative Example 1

[0116] The difference between Comparative Example 1 and Example 1 is that steps S2-S4 are removed, and the potato peptide-loaded nanoparticles in step S6 are replaced with an equal amount of potato peptide.

[0117] Comparative Example 2

[0118] The difference between Comparative Example 2 and Example 1 is that step S3 is removed, and the modified mesoporous silica in step S4.1 is replaced by an equal amount of amino-functionalized mesoporous silica.

[0119] Comparative Example 3

[0120] The difference between Comparative Example 3 and Example 1 is that step S5 is removed, and the mango leaf extract-loaded microspheres in step S6 are replaced with an equal amount of mango leaf extract.

[0121] Comparative Example 4

[0122] The difference between this comparative example 4 and example 1 is that step S5 is removed, and the mango leaf extract-loaded microspheres in step S6 are replaced by an equal amount of potato peptide-loaded nanoparticles.

[0123] Comparative Example 5

[0124] The difference between this comparative example 5 and example 1 is that steps S1-S4 are removed, and the potato peptide-loaded nanoparticles in step S6 are replaced by an equal amount of mango leaf extract-loaded microspheres.

[0125] Performance Testing

[0126] Test 1: The mass concentration of potato peptides in the filtrate after centrifugation in step S4.3 of Examples 1-3 and Comparative Example 2 was detected by high performance liquid chromatography. The loading rate of potato peptides on modified mesoporous silica was calculated as follows:

[0127] Loading rate (%) = [(m 0 -C×V) / m 0 ]×100%;

[0128] Wherein, R is the loading rate, C is the mass concentration of potato peptide in the filtrate, V is the volume of the filtrate, and m0 is the initial mass of potato peptide. The results are shown in Table 1.

[0129] Table 1: Comparison of the loading rates of potato peptides on modified mesoporous silica in Examples 1-3 and Comparative Example 2

[0130] Example 1 Example 2 Example 3 Comparative Example 2 Load rate (%) 78.6 78.8 79.1 63.2

[0131] Test 2: 25 SD rats were randomly divided into 5 groups, 5 rats in each group, and the potato peptide nutritional supplements prepared in Examples 1-3 and Comparative Examples 1 and 3 were respectively gavaged. The rats were fasted for 12 hours before gavage, and sufficient water was ensured. The gavage concentration of the 5 groups of rats was 50 mg / kg. Recording began after oral administration. Blood samples were obtained by orbital venous sinus puncture at 0.5h, 1.0h, 2.0h, 4.0h, 6.0h, 8.0h, 10.0h, 12.0h, and 24.0h, respectively. The blood samples were immediately centrifuged at 3000r / min for 10min. Then, 20μL of serum sample was accurately drawn and extracted with 100μL of anhydrous ethanol. The extraction system was shaken on a vortex agitator for 1min and then centrifuged at 10000r / min for 10min. Finally, the supernatant was taken for HPLC detection to determine the total concentration of potato peptides and mango leaf extracts in each sample. The blood drug concentration data were model fitted and analyzed by software DAS2.0, and the area under the drug-time curve was used as the bioavailability parameter. The results are shown in Table 2.

[0132] Table 2: Comparison of the area under the drug-time curve of Examples 1-3 and Comparative Examples 1 and 3

[0133] Area under the drug-time curve Example 1 123.86 Example 2 124.21 Example 3 123.79 Comparative Example 1 63.27 Comparative Example 3 109.34

[0134] It can be seen from Table 1 and Table 2 that when the modified mesoporous silica is not used to load the potato peptide in Comparative Example 1, the area under the drug-time curve of the potato peptide nutritional supplement prepared is much lower than that in Example 1, that is, the bioavailability of the potato peptide nutritional supplement prepared in Comparative Example 1 is lower than that in Example 1, and when the mesoporous silica is not modified in Comparative Example 2, the loading rate of the modified mesoporous silica for potato peptide is about 63.2%, which is much lower than that in Example 1. It can be seen that by first using 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide The taurocholic acid solution is activated by imide, and then mixed with ethylene glycol chitosan solution for coupling reaction to prepare a modifier, which is then used to modify amino-functionalized mesoporous silica. After the modified mesoporous silica is used to load potato peptides, on the one hand, the modifier can provide more active sites, thereby increasing the loading rate of mesoporous silica for potato peptides. On the other hand, due to the loading effect of the modified mesoporous silica, the potato peptides can be protected from the acidic environment and protease in gastric juice, thereby effectively improving the bioavailability of the potato peptides.

[0135] In addition, when the mango leaf extract is not loaded in Comparative Example 3, the area under the drug-time curve of the prepared potato peptide nutritional supplement is lower than that in Example 1, that is, the bioavailability of the potato peptide nutritional supplement prepared in Comparative Example 3 is lower than that in Example 1. It can be seen that by first dissolving the mango leaf extract in a mixture of lemongrass essential oil and coconut oil to prepare a mixed oil, and then fully mixing the mixed oil with a carboxymethyl chitosan aqueous solution, and then adding natural vanillin for cross-linking reaction to form a hydrogel, gel microspheres loaded with mango leaf extract are prepared. Since the carboxyl groups in the carboxymethyl chitosan molecules are deprotonated in an alkaline medium, the molecular chains are extended, and protonated under acidic conditions, the molecular chains shrink to maintain the stability of the gel, thereby protecting the mango leaf extract from gastric juice degradation, maintaining the activity of the mango leaf extract, and thereby improving its bioavailability.

[0136] Test 3: 50 μg of the potato peptide nutritional supplement prepared in Examples 1-3 and Comparative Examples 4-5 and 50 μL of α-glucosidase solution were mixed on a 96-well ELISA plate, reacted at room temperature for 30 minutes, and then 50 μL of 5 mmol / L p-nitrophenyl-D-pyranoglucoside solution was added. After reacting for 30 minutes, 100 μL of 0.2 mol / L sodium carbonate solution was added to terminate the reaction, and the absorbance was measured at 405 nm. The blank control group was replaced with distilled water instead of the α-glucosidase solution, and the control group was not added with the potato peptide nutritional supplement sample. The calculation formula for the α-glucosidase inhibition rate is as follows:

[0137] Inhibition rate (%) = [1-(potato peptide nutritional supplement group-blank group) / control group] × 100%. The results are shown in Table 3.

[0138] Table 3: Comparison of calculation results of α-glucosidase inhibition rate in Examples 1-3 and Comparative Examples 4-5

[0139] Inhibition rate (%) Example 1 86.7 Example 2 86.5 Example 3 86.9 Comparative Example 4 82.3 Comparative Example 5 78.6

[0140] As can be seen from Table 3, when the nutritional supplements prepared by using single potato peptide nanoparticles and single mango leaf extract microspheres in Comparative Examples 4-5 were used, the inhibition rates of α-glucosidase were 82.3% and 78.6%, respectively, which were both lower than those in Example 1. It can be seen that when potato peptide-loaded nanoparticles and mango leaf extract-loaded microspheres were mixed to prepare potato peptide nutritional supplements, since potato peptides and mango leaf extracts can inhibit the activity of α-glucosidase through different mechanisms, and potato peptides can improve insulin sensitivity, while mango leaf extract can promote insulin secretion, the combination of the two can interact with each other and synergistically promote the anti-diabetic effect of the nutritional supplement to more comprehensively regulate blood sugar levels.

[0141] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A method for preparing an anti-diabetic potato peptide nutritional supplement, characterized in that: The steps include: S1: Preparation of potato peptides The waste water from potato starch production is pretreated to obtain pretreated waste, which is then dispersed in distilled water, enzymatically hydrolyzed, and then fermented to obtain potato peptides; S2: Preparation of amino-functionalized mesoporous silica Mesoporous silica nanoparticles are added to toluene at a solid-liquid ratio of 1 g: (30-40) mL, and ultrasonically dispersed for 20-30 min to obtain a mesoporous silica nanoparticle suspension, and then 3-aminopropyltriethoxysilane is added to the mesoporous silica nanoparticle suspension at a volume ratio of 1: (140-150), heated at 100-120° C. with stirring and reflux reaction for 10-12 h, cooled, centrifuged, washed and dried to obtain amino-functionalized mesoporous silica; S3: Preparation of modified mesoporous silica Ethylene glycol chitosan and taurocholic acid are respectively prepared into an ethylene glycol chitosan solution and a taurocholic acid solution, then the taurocholic acid solution is activated, and then the activated taurocholic acid solution is added into the ethylene glycol chitosan solution for reaction to prepare a modifier, and finally, the amino-functionalized mesoporous silica is modified by the modifier to obtain modified mesoporous silica; S4: Preparation of potato peptide-loaded nanoparticles The potato peptide and the modified mesoporous silica are prepared into a potato peptide solution and a modified mesoporous silica suspension, and then the potato peptide solution is added into the modified mesoporous silica suspension for ultrasonic loading to obtain potato peptide-loaded nanoparticles; S5: Preparation of mango leaf extract loaded microspheres Lemongrass essential oil, coconut oil, mango leaf extract and Tween 80 are mixed to prepare a mixed oil liquid, then a carboxymethyl chitosan aqueous solution is mixed with the mixed oil liquid, natural vanillin is added to react, anhydrous ethanol is added, stirred to break the emulsion, centrifuged, washed and dried to obtain mango leaf extract loaded microspheres; S6: Preparation of potato peptide nutritional supplements The potato peptide-loaded nanoparticles and the mango leaf extract-loaded microspheres are fully and uniformly mixed at a mass ratio of (13-15):1 to obtain a potato peptide nutritional supplement.

2. The method for preparing an anti-diabetic potato peptide nutritional supplement according to claim 1, characterized in that: S1 specifically includes the following steps: S1.1: placing the waste residue and wastewater from potato starch production in a centrifuge for centrifugal separation to remove the wastewater, and then washing the waste residue to obtain pre-treated waste residue; S1.2: Add distilled water to the above pre-treated waste residue at a solid-liquid ratio of 1g: (8-10)mL, stir and disperse thoroughly, heat to 35-40°C and keep warm, then add composite protease, keep warm for 1-2h, inactivate the enzyme, and obtain the enzymatic pre-treatment stock solution; S1.3: Inoculate the mixed strains into the above enzymatic pretreatment stock solution, ferment at 32-36°C for 16-18h, sterilize at high temperature, centrifuge, concentrate by membrane filtration and freeze-dry to obtain potato peptides.

3. The method for preparing an anti-diabetic potato peptide nutritional supplement according to claim 2, characterized in that: S3 specifically includes the following steps: S3.1: dissolving ethylene glycol chitosan and taurocholic acid in dimethyl sulfoxide at a solid-liquid ratio of 1 g:(40-50) mL, respectively, to obtain ethylene glycol chitosan solution and taurocholic acid solution; S3.2: Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide to the above taurocholic acid solution at a solid-liquid ratio of 1 g:(1-2) g:(20-30) mL, and stir at a rate of 400-500 r / min for 2-3 h to activate and obtain an activated taurocholic acid solution; S3.3: adding an equal volume of the above ethylene glycol chitosan solution to the above activated taurocholic acid solution, stirring and reacting for 6-8 hours, dialyzing and freeze-drying to obtain a modifier; S3.4: The amino-functionalized mesoporous silica obtained in step S2 is uniformly dispersed in PBS buffer at a solid-liquid ratio of 1g:(35-40)mL, and then the above-mentioned modifier is added, stirred to react for 10-12h, and modified mesoporous silica is obtained by centrifugation, washing and drying, wherein the mass ratio of the modifier to the amino-functionalized mesoporous silica is 1:(115-125).

4. The method for preparing an anti-diabetic potato peptide nutritional supplement according to claim 3, characterized in that: S4 specifically includes the following steps: S4.1: dissolving the potato peptide prepared in step S1.3 in PBS buffer at a solid-liquid ratio of 1 g:(25-35) mL to obtain a potato peptide solution; S4.2: Add the modified mesoporous silica to PBS buffer at a solid-liquid ratio of 1 g: (45-55) mL, and disperse by ultrasonic for 10-20 min to obtain a modified mesoporous silica suspension; S4.3: Add the potato peptide solution to the modified mesoporous silica suspension in a volume ratio of (6-8): 1, stir for 10-12 hours, and then ultrasonically load for 20-30 minutes. After centrifugal separation, washing and drying, potato peptide-loaded nanoparticles are obtained.

5. The method for preparing an anti-diabetic potato peptide nutritional supplement according to claim 4, characterized in that: S5 specifically includes the following steps: S5.1: lemongrass essential oil and coconut oil are mixed in a volume ratio of 1:(26-30), and then mango leaf extract and Tween 80 are added, heated and stirred at 40-50° C. for 20-30 min, and mixed thoroughly to obtain a mixed oil; S5.2: adding 1-2 wt % carboxymethyl chitosan aqueous solution to an equal volume of the above mixed oil, stirring for 10-20 min, adding natural vanillin, stirring and reacting for 2-3 h to obtain a mixed emulsion; S5.3: Anhydrous ethanol was added to the mixed emulsion in a volume ratio of (3-4): 1, and the mixed emulsion was stirred and demulsified at a rate of 1500-1600 r / min in an ice water bath at 0°C for 20-30 min, and then centrifuged to retain the gel layer. After washing and drying, microspheres loaded with mango leaf extract were obtained.

6. The method for preparing an anti-diabetic potato peptide nutritional supplement according to claim 2, characterized in that: The composite protease is prepared by mixing trypsin and pepsin in a mass ratio of (1-3):1, and the added amount of the composite protease is 3-5% of the mass of the pre-treated waste residue.

7. The method for preparing an anti-diabetic potato peptide nutritional supplement according to claim 2, characterized in that: The mixed bacteria include lactic acid bacteria and yeast, and the added amount of the mixed bacteria is 10-12% of the enzymatic pretreatment original solution.

8. The method for preparing an anti-diabetic potato peptide nutritional supplement according to claim 5, characterized in that: The solid-liquid ratio of mango leaf extract to coconut oil is 1 g:(60-70) mL, and the volume ratio of Tween 80 to coconut oil is 1:(20-30).

9. The method for preparing an anti-diabetic potato peptide nutritional supplement according to claim 5, characterized in that: The solid-to-liquid ratio of natural vanillin to carboxymethyl chitosan aqueous solution is 1 g: (25-30) mL.

10. An anti-diabetic potato peptide nutritional supplement, characterized in that: The anti-diabetic potato peptide nutritional supplement is prepared by the preparation method of the anti-diabetic potato peptide nutritional supplement according to any one of claims 1 to 9.