Preparation method of coconut oligosaccharide for adjuvant therapy of hyperglycemia and hyperlipidemia

Abstract

The invention provides a preparation method of coconut oligosaccharide for adjuvant therapy of hyperglycemia and hyperlipidemia, which comprises the following steps: (1) dicing coconut meat, pulping, homogenizing at high pressure to obtain homogenate, and cooling and filtering to obtain primary filtrate; (2) adding a compound enzyme preparation into the primary filtrate for enzymolysis, centrifuging and collecting supernate to obtain secondary filtrate; (3) adding the secondary filtrate into ionic liquid for fractional extraction, collecting extract liquor, evaporating, and performing vacuum concentration to obtain concentrated liquor; and (4) adsorbing and decolorizing the concentrated solution through a macroporous adsorption resin column, eluting with deionized water, collecting a sugar-containing component, and freeze-drying to obtain the target coconut oligosaccharide. The coconut oligosaccharide obtained through the synergistic effect of the compound enzyme preparation and the ionic liquid is high in yield. The coconut oligosaccharide can inhibit the activity of alpha-glucosidase, has a good cholesterol binding capacity, also has a good inhibition rate on lipase, and has a good auxiliary treatment effect on hyperglycemia and hyperlipidemia.

Description

Technical Field

[0001] The invention relates to the technical field of oligosaccharide preparation, and in particular to a method for preparing coconut oligosaccharide for auxiliary treatment of hyperglycemia and hyperlipidemia. Background Art

[0002] Hyperglycemia and hyperlipidemia are common metabolic diseases in modern society, severely impacting human health and potentially leading to complications such as cardiovascular and cerebrovascular disease and diabetes. Currently, a wide variety of medications are available for treating these conditions, but long-term use can lead to side effects. Therefore, identifying safe and effective adjunctive treatments is of great practical significance.

[0003] Oligosaccharides, also known as oligosaccharides or oligosaccharides, refer to compounds composed of 2-10 glycosidic bonds. They are a new type of functional sugar source with multiple physiological functions, such as regulating intestinal flora, assisting in regulating blood sugar and blood lipids, promoting nutrient metabolism, and helping mineral absorption. Oligosaccharides integrate nutrition, health care, and diet therapy, and are widely used in food, health products, beverages, medicine, feed additives and other fields.

[0004] Coconut is a common tropical fruit with a rich carbohydrate content in its pulp. Currently, the deep-processed products of coconut are mainly coconut oil, coconut powder, coconut sugar, coconut water, etc. There are relatively few extraction methods for coconut oligosaccharides at present. There are shortcomings such as oligosaccharides being difficult to dissolve during the extraction process and low extraction yield. Therefore, a suitable extraction method for coconut oligosaccharides is sought to further expand the deep processing and utilization of coconut. Summary of the Invention

[0005] In view of this, the present invention provides a preparation method of coconut oligosaccharides for auxiliary treatment of hyperglycemia and hyperlipidemia.

[0006] The technical solution of the present invention is achieved as follows:

[0007] A method for preparing coconut oligosaccharides for assisting in the treatment of hyperglycemia and hyperlipidemia comprises the following steps:

[0008] (1) chopping coconut meat into pieces, mixing the pieces with water, and beating the pieces to obtain a slurry, subjecting the slurry to high-pressure homogenization to obtain a homogenate, and then cooling the homogenate at 2 to 4° C. for 6 to 8 hours and filtering the mixture to obtain a primary filtrate;

[0009] (2) adding a composite enzyme preparation to the primary filtrate for oscillating enzymolysis, and collecting the supernatant by centrifugation to obtain a secondary filtrate;

[0010] (3) adding the secondary filtrate to the ionic liquid for fractional extraction, collecting the extract, evaporating the extract, and concentrating it in vacuo to obtain a concentrate;

[0011] (4) The concentrated solution is decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components are collected and freeze-dried to obtain coconut oligosaccharides for auxiliary treatment of hyperglycemia and hyperlipidemia.

[0012] Furthermore, the mass ratio of the coconut meat to water is 3 to 5:1; and the high-pressure homogenization is performed 2 to 3 times under the condition of 30 to 50 MPa.

[0013] Furthermore, the added amount of the complex enzyme preparation is 2% to 4% of the mass of the primary filtrate.

[0014] Furthermore, the complex enzyme in the complex enzyme preparation includes cellulase, papain, pectinase and glucanase in a mass ratio of 7-10:2-3:2-4:3-5.

[0015] Furthermore, the preparation method of the complex enzyme preparation is: taking the complex enzyme, adding it to a phosphate buffer solution, stirring and mixing, then adding polyethylene glycol 400 while stirring to form a suspension, adding 1,8-bismaleimido-diglycol to the suspension, stirring and cross-linking in an ice water bath for 2 to 4 hours, centrifuging to collect the precipitate, and repeatedly washing the precipitate 3 to 4 times to obtain the complex enzyme preparation.

[0016] Furthermore, the concentration of the phosphate buffer solution is 18-20 mg / mL; the mass-to-volume ratio of the complex enzyme, phosphate buffer solution, and polyethylene glycol 400 is 1-2 mg:6-8 mL:20-30 mL.

[0017] Furthermore, the mass of the added 1,8-bismaleimide-diglycol is 2% to 3% of the mass of the suspension.

[0018] Furthermore, the centrifugal conditions are: rotation speed 3500-4000 r / min, time 20-25 min.

[0019] Furthermore, the volume ratio of the secondary filtrate to the ionic liquid is 5-7:1-2.

[0020] Furthermore, the preparation method of the ionic liquid is as follows: 1-ethyl-3-methylimidazole lactate is mixed with an ethanol solution; the mass concentration of the ethanol solution is 70% to 80%; and the volume ratio of the 1-ethyl-3-methylimidazole lactate to the ethanol solution is 3 to 5:1 to 2.

[0021] Furthermore, the oscillation enzymolysis conditions are: temperature of 45-55° C., pH of 6.0-7.5, oscillation speed of 100-150 r / min, and oscillation enzymolysis time of 60-300 min.

[0022] Furthermore, the pore size of the ultrafiltration membrane is 3000-5000 Da.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The present invention first solidifies, separates and removes lipid substances by cryogenic cooling during the extraction process of coconut oligosaccharides, and then effectively extracts coconut oligosaccharides through the synergistic effect of a complex enzyme preparation and an ionic liquid, and the prepared coconut oligosaccharides have a high yield. In addition, the coconut oligosaccharides prepared by the present invention are more easily absorbed, can inhibit the activity of α-glucosidase, reduce the absorption of glucose, have good cholesterol binding capacity, also have a good inhibition rate on lipase, can reduce the body's absorption of fat, and have a good auxiliary treatment effect for hyperglycemia and hyperlipidemia.

[0025] In the present invention, the composite enzyme preparation has high stability after precipitation cross-linking and can improve the activity of the composite enzyme. In addition, through the synergistic effect of the composite enzyme preparation and the ionic liquid, the destruction of coconut cell walls can be enhanced, the rate of sugar substances dissolving out of cells and the degradation of macromolecular polysaccharides into oligosaccharides can be accelerated, and impurities such as cellulose, protein and pectin can be removed, which is beneficial to the subsequent separation and purification of coconut oligosaccharides, effectively shortens the preparation time, maintains the activity of oligosaccharides, and also improves the yield of coconut oligosaccharides. DETAILED DESCRIPTION

[0026] In order to better understand the technical content of the present invention, specific examples are provided below to further illustrate the present invention.

[0027] Unless otherwise specified, the experimental methods used in the examples of the present invention are all conventional methods.

[0028] Unless otherwise specified, the materials, reagents, etc. used in the examples of the present invention can be obtained from commercial sources.

[0029] The cellulase of the present invention is produced by Nanning Pangbo Bioengineering Co., Ltd., and the specification is 20,000 u / g.

[0030] The papain of the present invention is produced by Nanning Pangbo Bioengineering Co., Ltd., and the specification is 20,000 u / g.

[0031] The pectinase of the present invention is produced by Nanning Pangbo Bioengineering Co., Ltd., and the specification is 30,000 u / g.

[0032] The manufacturer of the glucanase of the present invention is Nanning Pangbo Bioengineering Co., Ltd., and the specification is 30,000 u / g.

[0033] Example 1

[0034] The complex enzyme preparation includes cellulase, papain, pectinase and glucanase in a mass ratio of 7:3:4:3;

[0035] The preparation method of the complex enzyme preparation is as follows: taking the complex enzyme, adding it to a phosphate buffer solution with a concentration of 19 mg / mL and stirring to mix evenly, then adding polyethylene glycol 400 while stirring to prepare a suspension, adding 1,8-bismaleimido-diglycol to the suspension, stirring and cross-linking in an ice water bath for 2 hours, collecting the precipitate by centrifugation, and repeatedly washing the precipitate with a phosphate buffer solution with a concentration of 19 mg / mL for 4 times to obtain the complex enzyme preparation; the mass volume ratio of the complex enzyme, phosphate buffer, and polyethylene glycol 400 is 1 mg:8 mL:26 mL; the mass of 1,8-bismaleimido-diglycol added is 3% of the mass of the suspension.

[0036] The preparation method of the ionic liquid comprises the following steps: uniformly mixing 1-ethyl-3-methylimidazole lactate with an ethanol solution; the mass concentration of the ethanol solution is 75%; and the volume ratio of the 1-ethyl-3-methylimidazole lactate to the ethanol solution is 3:2.

[0037] A method for preparing coconut oligosaccharides for assisting in the treatment of hyperglycemia and hyperlipidemia comprises the following steps:

[0038] (1) The coconut meat is cut into pieces and mixed with water in a mass ratio of 4:1, and then beaten to obtain a slurry. The slurry is homogenized three times under 50 MPa to obtain a homogenate. The homogenate is then cooled at 4°C for 8 h and filtered to obtain a primary filtrate.

[0039] (2) adding a composite enzyme preparation to the primary filtrate and performing oscillation enzymolysis for 120 min at a temperature of 55°C, a pH of 7.5, and an oscillation speed of 100 r / min, then centrifuging for 20 min at a speed of 3800 r / min to collect the supernatant to obtain a secondary filtrate; the amount of the composite enzyme preparation added was 4% of the mass of the primary filtrate;

[0040] (3) adding the secondary filtrate to the ionic liquid in a volume ratio of 6:2, fractionating and extracting the secondary filtrate, collecting the extract, evaporating the extract, and concentrating in vacuo to obtain a concentrate;

[0041] (4) The concentrated solution was decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. The concentrated solution was then filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain a permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0042] Example 2

[0043] The complex enzyme preparation includes cellulase, papain, pectinase and glucanase in a mass ratio of 9:2:2:4;

[0044] The preparation method of the complex enzyme preparation is as follows: taking the complex enzyme, adding it to a phosphate buffer solution with a concentration of 18 mg / mL, stirring and mixing, then adding polyethylene glycol 400 while stirring to form a suspension, adding 1,8-bismaleimido-diglycol to the suspension, stirring and cross-linking in an ice water bath for 3 hours, centrifuging to collect the precipitate, and repeatedly washing the precipitate with a phosphate buffer solution with a concentration of 18 mg / mL for 3 times to obtain the complex enzyme preparation; the mass volume ratio of the complex enzyme, phosphate buffer, and polyethylene glycol 400 is 2 mg:6 mL:30 mL; the mass of 1,8-bismaleimido-diglycol added is 2% of the mass of the suspension.

[0045] The preparation method of the ionic liquid comprises the following steps: uniformly mixing 1-ethyl-3-methylimidazole lactate with an ethanol solution; the mass concentration of the ethanol solution is 70%; and the volume ratio of the 1-ethyl-3-methylimidazole lactate to the ethanol solution is 5:1.

[0046] A method for preparing coconut oligosaccharides for assisting in the treatment of hyperglycemia and hyperlipidemia comprises the following steps:

[0047] (1) The coconut meat is cut into pieces and mixed with water in a mass ratio of 5:1, and then beaten to obtain a slurry. The slurry is homogenized twice under 30 MPa to obtain a homogenate, and then the homogenate is cooled at 3°C ​​for 6 hours and filtered to obtain a primary filtrate;

[0048] (2) adding a composite enzyme preparation to the primary filtrate and performing oscillation enzymolysis for 150 min at a temperature of 45°C, a pH of 6.8, and an oscillation speed of 120 r / min, then centrifuging at a speed of 3500 r / min for 25 min to collect the supernatant to obtain a secondary filtrate; the amount of the composite enzyme preparation added was 2% of the mass of the primary filtrate;

[0049] (3) adding the secondary filtrate to the ionic liquid at a volume ratio of 7:1, fractionating and extracting the secondary filtrate, collecting the extract, evaporating the extract, and concentrating in vacuo to obtain a concentrate;

[0050] (4) The concentrated solution was decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. The concentrated solution was then filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain a permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0051] Example 3

[0052] The complex enzyme preparation includes cellulase, papain, pectinase and glucanase in a mass ratio of 10:3:3:5;

[0053] The preparation method of the complex enzyme preparation is as follows: taking the complex enzyme, adding it to a phosphate buffer solution with a concentration of 20 mg / mL, stirring and mixing, then adding polyethylene glycol 400 while stirring to form a suspension, adding 1,8-bismaleimido-diglycol to the suspension, stirring and cross-linking in an ice water bath for 3 hours, centrifuging to collect the precipitate, and repeatedly washing the precipitate with a phosphate buffer solution with a concentration of 20 mg / mL for 4 times to obtain the complex enzyme preparation; the mass volume ratio of the complex enzyme, phosphate buffer, and polyethylene glycol 400 is 2 mg:7 mL:20 mL; the mass of 1,8-bismaleimido-diglycol added is 2.5% of the mass of the suspension.

[0054] The preparation method of the ionic liquid comprises the following steps: uniformly mixing 1-ethyl-3-methylimidazole lactate with an ethanol solution; the mass concentration of the ethanol solution is 80%; and the volume ratio of the 1-ethyl-3-methylimidazole lactate to the ethanol solution is 4:1.

[0055] A method for preparing coconut oligosaccharides for assisting in the treatment of hyperglycemia and hyperlipidemia comprises the following steps:

[0056] (1) The coconut meat is cut into pieces and mixed with water in a mass ratio of 3:1, and then beaten to obtain a slurry. The slurry is homogenized three times under 40 MPa to obtain a homogenate. The homogenate is then cooled at 2° C. for 7 h and filtered to obtain a primary filtrate.

[0057] (2) adding a composite enzyme preparation to the primary filtrate and performing oscillation enzymolysis for 120 min at a temperature of 50° C., a pH of 7.2, and an oscillation speed of 150 r / min, and then centrifuging for 20 min at a speed of 4000 r / min to collect the supernatant to obtain a secondary filtrate; the amount of the composite enzyme preparation added was 3% of the mass of the primary filtrate;

[0058] (3) adding the secondary filtrate to the ionic liquid in a volume ratio of 5:2, fractionating and extracting the secondary filtrate, collecting the extract, evaporating the extract, and concentrating in vacuo to obtain a concentrate;

[0059] (4) The concentrated solution was decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. The concentrated solution was then filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain a permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0060] Comparative Example 1

[0061] The difference between this comparative example and Example 3 is that no complex enzyme preparation is added for oscillation enzymolysis, and the rest is consistent with Example 3.

[0062] The preparation method of coconut oligosaccharides for assisting the treatment of hyperglycemia and hyperlipidemia in this comparative example comprises the following steps:

[0063] (1) The coconut meat is cut into pieces and mixed with water in a mass ratio of 3:1, and then beaten to obtain a slurry. The slurry is homogenized three times under 40 MPa to obtain a homogenate. The homogenate is then cooled at 2° C. for 7 h and filtered to obtain a primary filtrate.

[0064] (2) adding the primary filtrate to the ionic liquid in a volume ratio of 5:2, fractionating and extracting the primary filtrate, collecting the extract, evaporating the extract, and concentrating in vacuo to obtain a concentrate;

[0065] (3) The concentrated solution was decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. The filtrate was then filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain a permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0066] Comparative Example 2

[0067] The difference between this comparative example and Example 3 is that no ionic liquid is added for fractional extraction, and the rest is consistent with Example 3.

[0068] The preparation method of coconut oligosaccharides for assisting the treatment of hyperglycemia and hyperlipidemia in this comparative example comprises the following steps:

[0069] (1) The coconut meat is cut into pieces and mixed with water in a mass ratio of 3:1, and then beaten to obtain a slurry. The slurry is homogenized three times under 40 MPa to obtain a homogenate. The homogenate is then cooled at 2° C. for 7 h and filtered to obtain a primary filtrate.

[0070] (2) adding a composite enzyme preparation to the primary filtrate and performing oscillatory enzymolysis for 120 min at a temperature of 50° C., a pH of 7.2, and an oscillation speed of 150 r / min, then centrifuging for 20 min at a speed of 4000 r / min to collect the supernatant to obtain a secondary filtrate; the amount of the composite enzyme preparation added is 3% of the mass of the primary filtrate; evaporating the supernatant and concentrating in vacuo to obtain a concentrate;

[0071] (3) The concentrated solution was decolorized by adsorption on a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. The filtrate was then filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain a permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0072] Comparative Example 3

[0073] The difference between this comparative example and Example 3 is that the preparation method of the complex enzyme preparation of the present invention is different from that of Example 3, that is, the complex enzyme of this comparative example is simply mixed evenly without precipitation and cross-linking, and the rest is consistent with Example 3.

[0074] The preparation method of the composite enzyme preparation of this comparative example is as follows: cellulase, papain, pectinase and glucanase are uniformly mixed in a mass ratio of 10:3:3:5.

[0075] Test example

[0076] The properties of the coconut oligosaccharides prepared by the preparation methods of Examples 1-3 and Comparative Examples 1-3 were measured.

[0077] (1) Yield of coconut oligosaccharides

[0078] The coconut oligosaccharides were prepared according to the preparation methods of Examples 1-3 and Comparative Examples 1-3, and the yield (%) of coconut oligosaccharides was determined by the phenol-sulfuric acid method using glucose as a standard.

[0079] (2) α-glucosidase inhibition ability

[0080] The coconut oligosaccharide obtained by the preparation method of above-mentioned embodiment 1-3 and comparative example 1-3 is respectively prepared into a sample solution with a concentration of 4mg / mL using a potassium phosphate buffer of 67mmol / L and a pH of 6.8, and then the inhibition rate of alpha-glucosidase activity is determined by colorimetry. 1.0mL of the above-mentioned sample solution is fully mixed with 2.0mL of the alpha-glucosidase having an enzyme activity concentration of 2U / mL, and after incubation at 37°C for 20min, 4.5mL of p-NPG with a concentration of 20mmol / L is added, and after continuing to incubate at 37°C for 30min, 7.0mL of sodium carbonate solution with a concentration of 2mol / L is finally added to terminate the reaction. In a control tube, the same amount of potassium phosphate buffer is used instead of sample as a blank group, and alpha-glucosidase is not added as a background group in the sample group, and acarbose is a positive control. The absorbance measured at 405nm is used to determine the alpha-glucosidase inhibitory ability. The alpha-glucosidase inhibition rate calculation formula is as follows:

[0081] α-glucosidase inhibition rate (%) = [1-(A1-A2) / A0] × 100

[0082] Where: A0: blank group test value; A1: sample group test value; A2: background group test value.

[0083] (3) Cholesterol binding capacity

[0084] Fresh egg yolk was mixed with 9 volumes of distilled water to form a homogeneous emulsion. 0.2 g of coconut oligosaccharides prepared in Examples 1-3 and Comparative Examples 1-3 were weighed and added to 25 mL of the emulsion. The mixture was shaken in a 37°C water bath for 2 h and then centrifuged at 4000 rpm for 20 min. The supernatant was collected and the absorbance at 550 nm was measured using the o-phthalaldehyde method. The cholesterol binding capacity was calculated as follows:

[0085] Cholesterol binding capacity (mg / g) = (M1-M2) / M

[0086] Where: M: constant weight sample mass, g; M1: cholesterol content in the supernatant after centrifugation, mg; M2: cholesterol content in the egg yolk emulsion before adsorption, mg.

[0087] (4) Lipase inhibition ability

[0088] The coconut oligosaccharides obtained in Examples 1-3 and Comparative Examples 1-3 were respectively prepared into sample solutions with a concentration of 4 mg / mL using distilled water; 40 μL of a 100 mg / mL lipase solution was taken and mixed evenly with 100 μL of the sample solution, 3.0 mL of a 100 mmol / L, pH 7.0 potassium phosphate buffer was added and reacted at 30° C. for 3 min, and then 20 μL of p-nitrophenyl acetate-dimethyl sulfoxide solution was added and reacted for 10 min. The absorbance was measured at 405 nm, and the phosphate buffer of the same concentration and amount was used instead of the sample solution as a control group to measure its absorbance at 405 nm. The lipase inhibition rate was then calculated using the following formula:

[0089] Lipase inhibition rate (%) = (A1-A2) / A1×100

[0090] Where: A1: test value of the control group; A2: test value of the sample group.

[0091] The above measurement results are shown in Table 1 below:

[0092] Table 1

[0093]

[0094] The above results show that, when preparing coconut oligosaccharides, the composite enzyme preparation and ionic liquid prepared by ad hoc method of the embodiment of the present invention 1-3 are combined, and the preparation yield of coconut oligosaccharides can be improved.Show that the present invention is high and can improve the activity of composite enzyme by composite enzyme preparation after being cross-linked by precipitation, and by the synergistic effect of composite enzyme preparation and ionic liquid, the destruction of coconut cell wall can be enhanced, the speed of carbohydrate dissolving cell and the acceleration of macromolecular polysaccharide degradation are oligosaccharides, and the impurities such as cellulose, protein and pectin can also be removed, it is conducive to the subsequent separation and purification of coconut oligosaccharides, effectively shortening the preparation time and improving yield.

[0095] In addition, the coconut oligosaccharides prepared by Examples 1-3 of the present invention have a better inhibitory effect on α-glucosidase than Comparative Examples 1-3, and can effectively reduce the absorption of glucose, thereby assisting in lowering blood sugar levels. In terms of cholesterol binding capacity and lipase inhibition rate, Examples 1-3 of the present invention can bind more cholesterol compared to Comparative Examples 1-3, and can also reduce the body's absorption of fat by inhibiting lipase activity, thereby achieving the purpose of assisting in lowering blood lipids.

[0096] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A preparation method of coconut oligosaccharides for auxiliary treatment of hyperglycemia and hyperlipidemia, characterized in that, The following steps are involved: (1) chopping coconut meat into pieces, mixing the pieces with water, and beating the pieces to obtain a slurry, subjecting the slurry to high-pressure homogenization to obtain a homogenate, and then cooling the homogenate at 2 to 4° C. for 6 to 8 hours and filtering the mixture to obtain a primary filtrate; (2) adding a composite enzyme preparation to the primary filtrate for oscillating enzymolysis, and collecting the supernatant by centrifugation to obtain a secondary filtrate; (3) adding the secondary filtrate to the ionic liquid for fractional extraction, collecting the extract, evaporating the extract, and concentrating it in vacuo to obtain a concentrate; (4) The concentrated solution is subjected to adsorption decolorization by a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components are collected. The concentrated solution is then filtered through an ultrafiltration membrane to obtain a permeate, which is then freeze-dried to obtain coconut oligosaccharides for auxiliary treatment of hyperglycemia and hyperlipidemia.

2. the preparation method of coconut oligosaccharide for auxiliary treatment of hyperglycemia and hyperlipidemia according to claim 1, is characterized in that, The mass ratio of the coconut meat to water is 3-5:1; and the high-pressure homogenization is performed 2-3 times under the condition of 30-50 MPa.

3. the preparation method of coconut oligosaccharide for auxiliary treatment of hyperglycemia, hyperlipidemia according to claim 1, is characterized in that, The added amount of the complex enzyme preparation is 2% to 4% of the mass of the primary filtrate.

4. the preparation method of coconut oligosaccharide for auxiliary treatment of hyperglycemia, hyperlipidemia according to claim 1, is characterized in that, The complex enzyme in the complex enzyme preparation comprises cellulase, papain, pectinase and glucanase in a mass ratio of 7-10:2-3:2-4:3-5.

5. the preparation method of coconut oligosaccharide for auxiliary treatment of hyperglycemia, hyperlipidemia according to claim 1, is characterized in that, The preparation method of the complex enzyme preparation comprises the following steps: adding the complex enzyme to a phosphate buffer solution and stirring the mixture, then adding polyethylene glycol 400 while stirring to form a suspension, adding 1,8-bismaleimido-diglycol to the suspension, stirring and cross-linking in an ice-water bath for 2 to 4 hours, collecting a precipitate by centrifugation, and repeatedly washing the precipitate 3 to 4 times to obtain the complex enzyme preparation.

6. the preparation method of coconut oligosaccharide for auxiliary treatment of hyperglycemia, hyperlipidemia according to claim 4, is characterized in that, The concentration of the phosphate buffer solution is 18-20 mg / mL; the mass-to-volume ratio of the complex enzyme, phosphate buffer, and polyethylene glycol 400 is 1-2 mg: 6-8 mL: 20-30 mL; and the mass of the added 1,8-bismaleimido-diglycol is 2%-3% of the mass of the suspension.

7. the preparation method of coconut oligosaccharide for auxiliary treatment of hyperglycemia and hyperlipidemia according to claim 1, is characterized in that, The centrifugal conditions are: rotation speed 3500-4000 r / min, time 20-25 min.

8. The preparation method of coconut oligosaccharides for auxiliary treatment of hyperglycemia and hyperlipidemia according to claim 1, wherein The volume ratio of the secondary filtrate to the ionic liquid is 5-7:1-2.

9. The preparation method of coconut oligosaccharides for auxiliary treatment of hyperglycemia and hyperlipidemia according to claim 1, wherein The preparation method of the ionic liquid comprises the following steps: uniformly mixing 1-ethyl-3-methylimidazole lactate with an ethanol solution; the mass concentration of the ethanol solution is 70% to 80%; and the volume ratio of the 1-ethyl-3-methylimidazole lactate to the ethanol solution is 3 to 5:1 to 2.

10. The preparation method of coconut oligosaccharides for auxiliary treatment of hyperglycemia and hyperlipidemia according to claim 1, characterized in that, The pore size of the ultrafiltration membrane is 3000-5000Da.

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