CN115160449B - Moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect, and preparation method and application thereof - Google Patents
Moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect, and preparation method and application thereof Download PDFInfo
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- CN115160449B CN115160449B CN202210833825.6A CN202210833825A CN115160449B CN 115160449 B CN115160449 B CN 115160449B CN 202210833825 A CN202210833825 A CN 202210833825A CN 115160449 B CN115160449 B CN 115160449B
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- moringa oleifera
- moringa
- oleifera leaf
- extract
- glycolipid
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- A—HUMAN NECESSITIES
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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Abstract
The invention discloses a moringa oleifera leaf polysaccharide extract with a function of regulating glycolipid absorption, and a preparation method and application thereof. The preparation method adopts the processes of liquid nitrogen crushing, cellulase enzymolysis, high-temperature extraction, yeast fermentation, low-temperature high-speed centrifugation, reduced pressure concentration, ethanol precipitation, low-temperature high-speed centrifugation, water adding and re-dissolution, freeze drying and the like to obtain the moringa oleifera leaf polysaccharide extract with the function of regulating the absorption of glycolipid. The total sugar content in the moringa oleifera leaf polysaccharide extract is more than 50%, the yield is more than 12%, the moringa oleifera leaf polysaccharide extract can effectively block glucose diffusion, adsorb cholate, inhibit the solubility of cholesterol in micelles, and has a good effect of regulating glycolipid absorption. The invention has simple extraction process, the whole process flow can meet the food-grade requirement, and the invention can be applied to the fields of common foods, health care products and the like.
Description
Technical Field
The invention belongs to the field of finish machining and high-value treatment of moringa leaves, and particularly relates to a moringa leaf polysaccharide extract with a glycolipid absorption regulating effect, and a preparation method and application thereof.
Background
With the improvement of the living standard of modern people, the population of Chinese obesity is gradually increased year by year. The plant polysaccharide has remarkable activity of reducing blood sugar and controlling lipid, and is an important food blood sugar and lipid-controlling factor. The moringa leaves contain rich saccharides, proteins and other nutrients, and the polysaccharides are main active ingredients for reducing blood sugar and controlling lipid.
The group adopts the processes of crushing and sieving, alkali treatment, cellulase enzymolysis, high-pressure homogenization, high-temperature extraction, low-temperature high-speed centrifugation, ultrafiltration separation concentration, protease enzymolysis, low-temperature high-speed centrifugation, secondary ultrafiltration separation concentration, freeze drying and the like to obtain the moringa oleifera leaf extract with hypoglycemic activity (patent application number: CN 201811006140.4), the total sugar content of the extract is more than 30 percent, and the animal level shows that the extract has good hypoglycemic and hypolipidemic effects, wherein the active ingredients are polysaccharide and protein peptide, so the polysaccharide accounts for not high proportion, and the preparation time of the method is more than 3 days; in addition, the research team adopts the processes of crushing, sieving, hot reflux of ethanol for removing impurities, water adding and homogenizing, ultrasonic treatment, cellulase enzymolysis, high-temperature extraction, centrifugal separation, reduced pressure concentration, ethanol fractional precipitation, water adding and re-dissolution, secondary reduced pressure concentration and freeze drying to obtain the moringa oleifera leaf extract with cholate adsorption capacity (patent application number: CN 201811536815.6), the preparation method can effectively extract polysaccharide with cholate adsorption capacity (hypolipidemic potential) in moringa oleifera leaves, the preparation time is longer than 3 days, the total sugar content of the extract is more than 50 percent, compared with the total sugar content obtained by the patent (patent application number: CN 201811006140.4), but the functions of the extracts prepared by the two patent application technologies are different, the extract with hypolipidemic activity is prepared by CN201811006140.4, and the extract with hypolipidemic potential is prepared by CN 201811536815.6. In addition, research team discloses an acetylated moringa oleifera leaf polysaccharide with the effect of reducing blood sugar and a preparation method thereof, wherein acetic anhydride reaction, acid treatment, dialysis, ethanol precipitation and freeze drying are adopted to obtain the acetylated moringa oleifera leaf polysaccharide with the effect of reducing blood sugar, the preparation method has high requirements on the used processing equipment, the processing cost is high, and the preparation time period is more than 3 days. In addition, the method for extracting the active ingredients of the moringa leaves in a sectionalized way and the beverage and the preparation method of the beverage containing the active ingredients of the moringa leaves are disclosed, wherein the moringa She Rebeng is dried, crushed, subjected to double-tank water immersion extraction (respectively cellulase enzymolysis and protease enzymolysis), supernatant fluid is subjected to fine filtration, ultrafiltration and spray drying to obtain a moringa leaf aqueous extract, ethanol extract of the moringa leaves is obtained by precipitation of an alcohol immersion lower layer, but the active ingredients of the moringa leaves are extracted in sections, and then a plurality of medicinal and edible homologous powders are added to prepare the beverage. The application of moringa oleifera leaf polysaccharide in preparing medicines and foods for preventing and treating alcoholic liver injury discloses a moringa oleifera leaf polysaccharide extract which is obtained by adopting a water extraction and alcohol precipitation method and combining the technological processes of high-speed centrifugation, deproteinization, AB-8 type macroporous adsorption resin column and the like, wherein the impurity removal technology relates to toxic reagents such as trichloromethane, the polysaccharide loss rate is high, the preparation time is longer than 4 days, and the processing cost is high.
In summary, (1) the conventional grinding and sieving are the most traditional and most classical pretreatment technologies for extracting moringa oleifera leaf polysaccharide, but the insufficient pretreatment of the raw materials (especially the leaf raw materials with high fiber content) by the conventional grinding leads to the difficulty of breaking the upper limit of the polysaccharide extraction rate; (2) The common extraction means such as a high-temperature high-pressure water extraction method which is repeated for a long time and a plurality of times have high requirements on equipment performance and low production efficiency; ethanol precipitation cannot completely remove a large amount of residual monosaccharides and oligosaccharides, and the purity of the polysaccharide is not high; (3) Less reports of moringa oleifera leaf polysaccharides are concerned about regulating the sugar and lipid absorption functions; (4) Most extraction technologies aim at high purity and high activity, and the extraction efficiency and yield are ignored due to heavy purity and heavy activity.
Therefore, the invention adopts liquid nitrogen to fully crush the moringa leaves, adopts the technical means of enzymatic wall breaking and high-temperature extraction to efficiently extract moringa leaf polysaccharide, adopts the method of fermenting and consuming a large amount of reducing sugar and partial protein by dry yeast with high sugar-resistant activity and coupling ethanol precipitation to efficiently purify the moringa leaf polysaccharide, and the obtained moringa leaf polysaccharide extract has the capacity of adjusting sugar and lipid absorption. The technology has the advantages of high extraction efficiency, high extraction yield, good functional activity, high purity, low requirement on equipment processing performance and capability of realizing continuous production of 6 dimensions, and has potential of industrialized popularization and application.
Disclosure of Invention
The invention aims to provide a moringa oleifera leaf polysaccharide extract with a function of regulating the absorption of glycolipid, and a preparation method and application thereof. The total sugar content in the moringa oleifera leaf polysaccharide extract is more than 50%, the yield is more than 12%, and the extract has good effects of reducing blood sugar and controlling lipid through the INFOGEST static digestion model, so that the moringa oleifera leaf polysaccharide extract has the potential of being applied to controlling the absorption of glycolipid into health foods. The invention has simple extraction process, the whole process flow can meet the food-grade requirement, and the invention can be applied to the fields of common foods, health care products and the like.
The technical scheme of the invention is as follows:
the invention provides a preparation method of moringa oleifera leaf polysaccharide extract with a function of regulating glycolipid absorption. The moringa oleifera leaf polysaccharide extract with good glycolipid absorption regulation function is prepared from moringa oleifera leaf serving as a raw material through the processes of liquid nitrogen crushing, cellulase enzymolysis, high-temperature extraction, yeast fermentation, low-temperature high-speed centrifugation, reduced-pressure concentration, ethanol precipitation, low-temperature high-speed centrifugation, water adding and re-dissolution, freeze drying and the like. The total sugar content in the moringa oleifera leaf polysaccharide extract is more than 50%, the yield is more than 12%, the moringa oleifera leaf polysaccharide extract can effectively block glucose diffusion, adsorb cholate and inhibit the solubility of cholesterol in micelles, has good effect of regulating the absorption of glycolipid, the glucose dialysis delay index is more than 37%, and the cholate adsorption rate and the cholesterol micelle dissolution inhibition rate are more than 50%. The extraction process is simple, the whole process flow can meet the food-grade requirement, and the method can be applied to the fields of common foods, health care products and the like. The preparation method comprises the following steps:
(1) Crushing by liquid nitrogen: crushing the dried moringa leaves into dry powder by a liquid nitrogen crushing device to obtain moringa leaf dry powder;
(2) Enzymatic hydrolysis of cellulase: uniformly mixing the moringa leaf dry powder obtained in the step (1) with water, regulating the pH, adding cellulase after fully and uniformly mixing, and stirring at constant temperature for enzymolysis to obtain a suspension 1;
(3) Heating and extracting: heating and extracting the suspension 1 to obtain a suspension 2;
(4) And (3) yeast fermentation: adding active dry yeast into the suspension 2, and fermenting at constant temperature to obtain a suspension 3;
(5) Centrifuging and concentrating: centrifuging the suspension 3, and concentrating the supernatant under reduced pressure to obtain an extract 1;
(6) Ethanol precipitation: adding ethanol solution into the extract 1, fully and uniformly mixing to obtain mixed solution, standing, centrifuging, adding water into the precipitate, heating for fully dissolving, and concentrating under reduced pressure to obtain extract 2;
(7) And (3) freeze drying: and (3) freeze-drying the extracting solution 2 to obtain the moringa oleifera leaf polysaccharide extract.
Further, in the step (1), the condition of liquid nitrogen pulverization is that the pulverization is carried out at-150 to-120 ℃.
Further, in the step (2), the feed liquid ratio of the moringa oleifera leaf dry powder to water is 1:8-1:12 g/mL; adjusting the pH value to 4.0-5.0 by using hydrochloric acid, adding cellulose with the mass of 0.25-0.75% (w/w, g/g) of the moringa oleifera leaf dry powder obtained in the step (1), stirring at the speed of 120-180 r/min, and performing enzymolysis at the temperature of 45-55 ℃ for 1-3 h.
Further, in the step (3), the temperature of the heating extraction is 90-120 ℃, and the time of the heating extraction is 20-40 min.
Further, in the step (3), the temperature of the heating extraction is 100-110 ℃.
Further, in the step (4), the mass ratio of the added active dry yeast to the water in the step (2) is 1g (200-1000) mL, the fermentation temperature is 26-30 ℃, the fermentation time is 1-3 h, and the rotating speed is 110-130 rpm.
Further, in the step (5), the temperature of the centrifugation is 4-8 ℃, the centrifugal force of the centrifugation is 6000-8000 g, and the time of the centrifugation is 15-25 min. The temperature of the reduced pressure concentration is 55-65 ℃, the solid content in the extracting solution 1 is 4-10 wt%,
further, in the step (6), the volume of the added ethanol solution accounts for 70% -90% (v/v) of the total volume of the mixed solution, the standing temperature is 4-8 ℃, the standing time is 2-4 h, the centrifuging temperature is 4-8 ℃, the centrifuging force is 4000-8000 g, and the centrifuging time is 15-25 min. The mass of the added water is 100-150 times of the mass of the sediment, the temperature of the added water is 50-60 ℃, the time of the added water is 1-3 hours, the temperature of the decompressed concentration is 55-65 ℃, the solid content in the extracting solution 2 is 2-4 wt%, and the moringa oleifera leaf polysaccharide extract is obtained after freeze drying.
The invention provides a moringa oleifera leaf polysaccharide extract with a function of regulating glycolipid absorption, which is prepared by the preparation method.
The invention also provides application of the moringa oleifera leaf polysaccharide extract with the function of regulating the absorption of glycolipid in preparation of foods, health products or medicines with the function of reducing blood glucose and controlling the lipid.
The invention has the following advantages and effects:
(1) According to the invention, the acid method treatment and the cellulose wall breaking method are combined with high-temperature extraction, so that the dissolution of active ingredients is greatly promoted, the high Wen Yike has the double effects of enzyme deactivation and sterilization, and the subsequent addition of the high-sugar-resistance active dry yeast effectively consumes small molecular saccharides and part of proteins in the extracting solution, so that the effect of double improvement of the activity and the polysaccharide purity is achieved, the preparation time is less than 2 days, the total sugar content in the moringa oleifera leaf polysaccharide extract can be improved, and the purpose of rapidly preparing the high-activity moringa oleifera leaf polysaccharide extract can be achieved.
(2) The whole preparation process flow meets the food-grade requirement.
(3) The yield of the moringa oleifera leaf polysaccharide extract obtained by the invention is more than 12%, and the total sugar content is more than 50%. The preparation method is simple and quick, the extract has glucose diffusion blocking activity and strong cholate adsorption activity, can effectively inhibit the solubility of cholesterol in micelles, inhibit the excessive absorption of saccharides and lipids in gastrointestinal tracts, can improve the absorption of glycolipids in multiple ways, and can be applied to the fields of common foods, health-care products and the like.
Drawings
Fig. 1 is a particle size distribution diagram of moringa oleifera leaf powder.
Fig. 2 is a microstructure of moringa oleifera leaf powder under an optical microscope.
FIG. 3 is a bar graph of yields of moringa oleifera leaf aqueous extracts AE1 and AE2 of comparative examples 3-4, moringa oleifera leaf extracts ME1 and ME2 of comparative examples 5-6, and moringa oleifera leaf polysaccharide extracts MEA, MEB, and MEC of examples 1-3.
FIG. 4 is a bar graph of total sugar content in aqueous moringa leaf extracts AE1 and AE2 of comparative examples 3-4, moringa leaf extracts ME1 and ME2 of comparative examples 5-6, and moringa leaf polysaccharide extracts MEA, MEB, and MEC of examples 1-3.
FIG. 5 is a bar graph of the effect of dry moringa leaf powder M1 and M2 of comparative examples 1-2, moringa leaf aqueous extracts AE1 and AE2 of comparative examples 3-4, moringa leaf extracts ME1 and ME2 of comparative examples 5-6, and moringa leaf polysaccharide extracts MEA, MEB and MEC of examples 1-3 on glucose diffusion.
FIG. 6 is a bar graph of bile acid adsorption capacity of dry moringa oleifera leaf powder M1 and M2 of comparative examples 1-2, moringa oleifera leaf aqueous extracts AE1 and AE2 of comparative examples 3-4, moringa oleifera leaf extracts ME1 and ME2 of comparative examples 5-6, and moringa oleifera leaf polysaccharide extracts MEA, MEB and MEC of examples 1-3.
FIG. 7 is a bar graph of cholesterol micelle solubility inhibition capacity for dry moringa leaf powders M1 and M2 of comparative examples 1-2, moringa leaf aqueous extracts AE1 and AE2 of comparative examples 3-4, moringa leaf extracts ME1 and ME2 of comparative examples 5-6, and moringa leaf polysaccharide extracts MEA, MEB, and MEC of examples 1-3.
Detailed description of the preferred embodiments
For a better understanding of the present invention, the operation of the InFOGEST static digestion model will be specifically elucidated below and further described in connection with specific examples, but embodiments of the present invention are not limited thereto.
1. The INFOGEST static digestion model evaluates glucose control lipid lowering activity:
(1) Simulated oral liquid: comprises 15.1mmol/L KCl and 3.7mmol/L KH 2 PO 4 、13.6mmol/L NaHCO 3 、0.15mmol/L MgCl 2 、0.06mmol/L(NH 4 ) 2 CO 3 1.1mmol/L HCl and 1.5mmol/L CaCl 2 The solvents were all water.
(2) Simulating gastric juice: comprises 6.9mmol/L KCl and 0.9mmol/L KH 2 PO 4 、25mmol/L NaHCO 3 、47.2mmol/L NaCl、0.12mmol/L MgCl 2 、0.5mmol/L(NH 4 ) 2 CO 3 15.6mmol/L HCl and 0.15mmol/L CaCl 2 The solvents were all water.
(3) Simulation of intestinal juice: comprises 6.8mmol/L KCl and 0.8mmol/L KH 2 PO 4 、85mmol/L NaHCO 3 、38.4mmol/L NaCl、0.33mmol/L MgCl 2 8.4mmol/L HCl and 0.6mmol/L CaCl 2 The solvents were all water.
The digestion process of the samples in the info GEST model is as follows:
(1) Oral digestion: sampling, dissolving in 2mL simulated oral liquid, adjusting pH to 7.0 with deionized water and 5mol/LNaOH aqueous solution to make the final volume of oral digest be 2.5mL, and oscillating at 37deg.C for 2min.
(2) Gastric digestion: 2mL simulated gastric fluid and 7.50. Mu.L 0.3mmol/L CaCl were added to the oral digest 2 The aqueous solution was dissolved in 40mg of pepsin (final concentration 2000U/mL) with 0.17mL of simulated gastric fluid, added to the digest, and the pH was adjusted to 3.0 with deionized water and 5mol/L aqueous HCl to give a final volume of 5mL of gastric digest, and the gastric digest was shaken at constant temperature of 37℃for 2h.
(3) Small intestine digestion: 4mL simulated intestinal fluid and 10. Mu.L 0.3mmol/L CaCl were added to the stomach digest 2 In the aqueous solution, 5mg of pancreatin (final concentration 100U/mL) was dissolved in 1.25mL of simulated intestinal fluid, and then added to the digest, 66.4mg of pig bile salt was added to 0.75mL of simulated intestinal fluid, and after dissolution, the digest was added to the pH of the mixture, and deionized water and 5mol/LNaOH aqueous solution were used to adjust the pH to 7.0, so that the final volume of the digest was 10mL, and the mixture was subjected to constant temperature shaking at 37℃for 2 hours.
1.1 glucose blocking Capacity determination step:
180mg glucose and 0.1g of sample (concentration 10mg/mL in final 10mL digestion volume) were taken, simulated oral digestion and simulated gastric digestion were performed as described above, all of the digestion solution was transferred to a dialysis bag with a molecular weight cut-off of 3kDa before the start of the last step of simulated intestinal digestion, the dialysis bag was placed in a container with 100mL simulated intestinal fluid, after shaking dialysis at 37℃for 2 hours, 2mL of dialysis bag external fluid was collected, and the glucose content of the dialysis bag external fluid was determined using a glucose kit. The blank was treated in the same way as the sample group without the addition of sample. The delay index (%) of glucose diffusion from the dialysis bag into the dialysate is calculated as follows:
wherein the unit of glucose amount is mmol/L.
1.2 procedure for determining adsorption Capacity of bile acid salt:
preparation of cholesterol micelle solution: 3.125g of pig bile salt, 386.65mg of cholesterol and 706.15mg of oleic acid are dissolved by using 150mL of simulated intestinal fluid, ultrasonic emulsification is carried out for 2 hours until the cholesterol is completely dissolved, deionized water is used for fixing the volume to 200mL, the gel bundles are filled in a closed container, and the gel bundles are stored at 37 ℃ for 24 hours for use.
A sample of 0.05g (5 mg/mL in the final 10mL digestion volume) was taken and simulated oral digestion and simulated gastric digestion were performed as described above. Dissolving 5mg of pancreatin in 4mL of cholesterol micelle solution before digestion of simulated small intestine, and adding the mixed solution into simulated intestinal juice after uniform mixing; then 0.01mL of 0.3mmol/L CaCl is added 2 An aqueous solution; adjusting pH to 7.0 with deionized water and 5mol/L NaOH aqueous solution to give digestion volume of 10mL, oscillating at 37deg.C for 2h, ice-bathing for 15min, centrifuging at 8000g at 4deg.C for 15min, collecting supernatant, and measuring cholate content. The blank was treated in the same way as the sample group without the addition of sample. The cholate adsorption (%) was calculated according to the following formula:
wherein, C blank is the concentration of cholate in blank control group, and the unit is mmol/L; c the sample is the concentration of cholate in the sample group, and the unit is mmol/L.
1.3 cholesterol micelle dissolution inhibition ability measurement step:
a sample of 0.05g (5 mg/mL in the final 10mL digestion volume) was taken and simulated oral digestion and simulated gastric digestion were performed as described above. Dissolving 5mg of pancreatin in 4mL of cholesterol micelle solution before digestion of simulated small intestine, and adding the mixed solution into simulated intestinal juice after uniform mixing; then 0.01mL of 0.3mmol/L CaCl is added 2 An aqueous solution; adjusting pH to 7.0 with deionized water and 5mol/L NaOH aqueous solution to obtain a final digestion volume of 10mL, oscillating at 37deg.C for 2h, ice-bathing for 15min, centrifuging at 8000g and 4deg.C for 15min, collecting the digestion supernatant, adding anhydrous methanol at a ratio of 1:1 (v/v), repeatedly extracting the supernatant twice, and measuring cholesterol content by OPA method. The blank was treated in the same way as the sample group without the addition of sample. Cholesterol solubility inhibition (%) was calculated according to the following formula:
wherein, C blank is cholesterol concentration in μg/mL in blank control group; c the concentration of cholesterol in the sample group in μg/mL.
The glucose blocking ability, cholate adsorption ability and cholesterol micelle solubility inhibition ability of the moringa oleifera leaf extracts obtained in examples and comparative examples were measured using the above methods.
Example 1
A preparation method of Moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect comprises the following steps:
(1) Crushing by liquid nitrogen: 30g of dry moringa leaves are put into a liquid nitrogen crushing device at the temperature of-150 ℃ and crushed into dry powder to obtain moringa leaf dry powder M1A.
(2) Enzymatic hydrolysis of cellulase: uniformly mixing the moringa oleifera leaf dry powder M1A obtained in the step (1) with deionized water according to the proportion of 1:8g/mL, adding hydrochloric acid to adjust the pH to 4.0, adding cellulase after fully uniformly mixing, wherein the mass of the cellulase is 0.25% (w/w, g/g) of the moringa oleifera leaf dry powder M1A obtained in the step (1), and stirring at a constant temperature at 45 ℃ at a rotating speed of 120r/min for enzymolysis for 1h to obtain a suspension S1A.
(3) High-temperature extraction: extracting suspension S1A at 90deg.C for 20min to obtain suspension S2A.
(4) And (3) yeast fermentation: cooling the suspension S2A to room temperature, adding active dry yeast into the suspension S2A, fermenting at a constant temperature for 1h at 26 ℃ at a rotating speed of 110rpm, wherein the volume ratio of the mass of the added active dry yeast to the deionized water in the step (2) is 1g to 1000mL, and obtaining the suspension S3A.
(5) Centrifuging and concentrating: centrifuging suspension S3A at 4deg.C at 6000g for 15min, collecting supernatant, and concentrating under reduced pressure at 55deg.C until the solid content in the final concentrate is 4wt% to obtain extract E1A.
(6) Ethanol precipitation: adding absolute ethyl alcohol into the extract E1A to obtain a mixed solution, adding absolute ethyl alcohol with the volume accounting for 70% (v/v) of the total volume of the mixed solution, fully mixing at 4 ℃, standing for 2h, centrifuging at 4000g at 4 ℃ for 15min, taking out the lower layer of sediment, adding deionized water with the mass 100 times of that of the sediment, heating at 50 ℃ for fully dissolving for 1h, and concentrating under reduced pressure at 55 ℃ until the solid content in the final concentrated solution is 2wt%, thereby obtaining the extract E2A.
(7) And (3) freeze drying: and (3) freeze-drying the extracting solution E2A to obtain the moringa oleifera leaf polysaccharide extract MEA.
Example 2
A preparation method of Moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect comprises the following steps:
(1) Crushing by liquid nitrogen: 30g of dry moringa leaves are put into a liquid nitrogen crushing device at the temperature of minus 135 ℃ and crushed into dry powder to obtain moringa leaf dry powder M1.
(2) Enzymatic hydrolysis of cellulase: uniformly mixing the moringa oleifera leaf dry powder M1 obtained in the step (1) with deionized water according to the proportion of 1:10g/mL, adding hydrochloric acid to adjust the pH to 4.5, fully uniformly mixing, adding cellulase, wherein the mass of the cellulase is 0.50% (w/w, g/g) of the moringa oleifera leaf dry powder M1 obtained in the step (1), and stirring at the constant temperature of 50 ℃ at the rotating speed of 150r/min for enzymolysis for 2 hours to obtain a suspension S1B.
(3) High-temperature extraction: extracting suspension S1B at 105deg.C for 30min to obtain suspension S2B.
(4) And (3) yeast fermentation: cooling the suspension S2B to room temperature, adding active dry yeast into the suspension S2B, fermenting at constant temperature for 2h at 28 ℃ at a rotating speed of 120rpm, wherein the volume ratio of the mass of the added active dry yeast to the deionized water in the step (2) is 1g:333mL, and obtaining the suspension S3B.
(5) Centrifuging and concentrating: after the suspension S3B was centrifuged at 7000g for 20min at 6℃at high speed, the supernatant was concentrated under reduced pressure at 60℃until the solid content in the final concentrate became 7wt%, giving extract E1B.
(6) Ethanol precipitation: adding absolute ethyl alcohol into the extract E1B to obtain a mixed solution, adding absolute ethyl alcohol with the volume of 80% (v/v) of the total volume of the mixed solution, fully mixing at 6 ℃ and standing for 3h, centrifuging at 6000g speed for 20min at 6 ℃, taking out the lower precipitate, adding deionized water with the mass being 125 times of that of the precipitate, heating at 55 ℃ for fully dissolving for 2h, and concentrating under reduced pressure at 60 ℃ until the solid content in the final concentrated solution is 3wt%, thereby obtaining the extract E2B.
(7) And (3) freeze drying: and (3) freeze-drying the extracting solution E2B to obtain a moringa oleifera leaf polysaccharide extract MEB.
Example 3
A preparation method of Moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect comprises the following steps:
(1) Crushing by liquid nitrogen: 30g of dry moringa leaves are put into a liquid nitrogen crushing device at the temperature of minus 120 ℃ and crushed into dry powder to obtain moringa leaf dry powder M1C.
(2) Enzymatic hydrolysis of cellulase: uniformly mixing the moringa oleifera leaf dry powder M1C obtained in the step (1) with deionized water according to the proportion of 1:12g/mL, adding hydrochloric acid to adjust the pH to 5.0, adding cellulase after fully uniformly mixing, wherein the mass of the cellulase is 0.75% (w/w, g/g) of the moringa oleifera leaf dry powder M1C obtained in the step (1), and stirring at a constant temperature of 55 ℃ at a rotating speed of 180r/min for enzymolysis for 3 hours to obtain a suspension S1C.
(3) High-temperature extraction: extracting suspension S1C at 120deg.C for 40min to obtain suspension S2C.
(4) And (3) yeast fermentation: cooling the suspension S2C to room temperature, adding active dry yeast into the suspension S2C, fermenting at a constant temperature for 3h at a constant speed of 130rpm at 30 ℃ with the volume ratio of the mass of the added active dry yeast to the deionized water in the step (2) being 1g to 200mL, and obtaining the suspension S3C.
(5) Centrifuging and concentrating: centrifuging the suspension S3C at 8deg.C at 8000g for 25min, collecting supernatant, and concentrating under reduced pressure at 65deg.C until the solid content in the final concentrate is 10wt% to obtain extract E1C.
(6) Ethanol precipitation: adding absolute ethyl alcohol into the extract E1C to obtain a mixed solution, adding absolute ethyl alcohol with the volume accounting for 90% (v/v) of the total volume of the mixed solution, fully mixing at 8 ℃ and standing for 4 hours, centrifuging at 8000g of rotation speed at 8 ℃ for 25 minutes, taking out the lower-layer sediment, adding deionized water with the mass 150 times of that of the sediment, heating at 60 ℃ for fully dissolving for 3 hours, and concentrating under reduced pressure at 65 ℃ until the solid content in the final concentrated solution is 4wt%, thereby obtaining the extract E2C.
(7) And (3) freeze drying: and (3) freeze-drying the extracting solution E2C to obtain a moringa oleifera leaf polysaccharide extract MEC.
Comparative example 1
A preparation method of moringa oleifera leaf dry powder specifically comprises the following steps:
30g of dry moringa leaves are put into a liquid nitrogen crushing device at the temperature of minus 135 ℃ and crushed into dry powder to obtain moringa She Yedan crushed dry powder M1.
Comparative example 2
A preparation method of moringa oleifera leaf dry powder specifically comprises the following steps:
pulverizing 30g of dry Moringa oleifera leaves, and sieving with 40 mesh sieve to obtain powder M2.
Comparative example 3
A preparation method of moringa oleifera leaf water extract with glycolipid absorption regulating effect comprises the following steps:
30g of dry moringa leaves are put into a liquid nitrogen crushing device at the temperature of minus 135 ℃ and crushed into dry powder to obtain moringa leaf dry powder M1. Mixing M1 and deionized water uniformly at a ratio of 1:10g/mL, extracting at a rotating speed of 120r/min for 1h at 60 ℃, centrifuging at a rotating speed of 7000g for 20min at 6 ℃, taking supernatant, concentrating at 60 ℃ under reduced pressure until the solid content in the final concentrated solution is 7wt%, adding absolute ethyl alcohol to obtain a mixed solution, adding absolute ethyl alcohol with a volume of 80% (v/v) of the total volume of the mixed solution, fully mixing at 6 ℃, standing for 3h, centrifuging at a rotating speed of 6000g for 20min at 6 ℃, taking a lower precipitate, adding deionized water with a mass of 125 times of the precipitate, heating at 55 ℃ for fully dissolving for 2h, concentrating at 60 ℃ under reduced pressure until the solid content in the final concentrated solution is 3wt%, and carrying out freeze drying treatment to obtain the moringa oleifera leaf aqueous extract AE1.
Comparative example 4
A preparation method of moringa oleifera leaf water extract with glycolipid absorption regulating effect comprises the following steps:
pulverizing 30g of dry moringa leaves, sieving with a 40-mesh sieve to obtain moringa leaf dry powder M2, uniformly mixing M2 with deionized water at a ratio of 1:10g/mL, extracting at a rotating speed of 120r/min for 1h at 60 ℃, centrifuging at a rotating speed of 7000g for 20min at 6 ℃, collecting supernatant, concentrating under reduced pressure at 60 ℃ until the solid content in the final concentrate is 7wt%, adding absolute ethyl alcohol to obtain a mixed solution, adding absolute ethyl alcohol, wherein the volume of the absolute ethyl alcohol accounts for 80% (v/v) of the total volume of the mixed solution, fully mixing at 6 ℃, standing for 3h, centrifuging at a rotating speed of 6000g for 20min at 6 ℃, taking down precipitate, adding deionized water with a mass of 125 times of precipitate, heating at 55 ℃ for fully dissolving for 2h, concentrating under reduced pressure at 60 ℃ until the solid content in the final concentrate is 3wt%, and performing freeze drying treatment to obtain moringa leaf water extract AE2.
Comparative example 5
A preparation method of Moringa oleifera leaf extract with glycolipid absorption regulating effect comprises the following steps:
30g of dry moringa leaves are put into a liquid nitrogen crushing device at the temperature of minus 135 ℃ and crushed into dry powder to obtain moringa leaf dry powder M1. Uniformly mixing the moringa dry powder M1 with deionized water according to the proportion of 1:10g/mL, adding hydrochloric acid to adjust the pH to 4.5, fully mixing, adding cellulase, wherein the mass of the cellulase is 0.50% (w/w, g/g) of the mass of the moringa dry powder M1, stirring and hydrolyzing for 2 hours at the constant temperature of 150r/min, centrifuging at the high speed of 7000g at the temperature of 6 ℃ for 20 minutes, concentrating the supernatant at the reduced pressure of 60 ℃ until the solid content in the final concentrate is 7wt%, adding absolute ethyl alcohol to obtain a mixed solution, fully mixing the mixed solution at the temperature of 6 ℃, standing for 3 hours after fully mixing the mixed solution, centrifuging at the high speed of 6000g at the temperature of 6 ℃, adding deionized water with the mass of 125 times of sediment, heating and fully dissolving for 2 hours at the temperature of 55 ℃, concentrating the reduced pressure until the solid content in the final concentrate is 3wt%, and drying to obtain the moringa extract ME1.
Comparative example 6
A preparation method of Moringa oleifera leaf extract with glycolipid absorption regulating effect comprises the following steps:
30g of dry moringa leaves are put into a liquid nitrogen crushing device at the temperature of minus 135 ℃ and crushed into dry powder to obtain moringa leaf dry powder M1. Uniformly mixing the moringa dry powder M1 with deionized water according to the proportion of 1:10g/mL, adding hydrochloric acid to adjust the pH to 4.5, fully mixing, adding cellulase, wherein the mass of the cellulase is 0.50% (w/w, g/g) of the moringa dry powder M1, stirring and hydrolyzing for 2 hours at the constant temperature of 150r/min at the temperature of 50 ℃, heating and extracting for 30 minutes at the temperature of 105 ℃, centrifuging for 20 minutes at the high speed of 7000g at the temperature of 6 ℃, concentrating the supernatant at the reduced pressure of 60 ℃ until the solid content in the final concentrated solution is 7wt%, adding absolute ethyl alcohol to obtain a mixed solution, standing for 3 hours after fully mixing at the temperature of 6 ℃, centrifuging for 20 minutes at the high speed of 6000g at the temperature of 6 ℃, taking precipitate, adding deionized water with the mass of 125 times of the precipitate, heating and fully dissolving for 2 hours at the temperature of 55 ℃, concentrating at the reduced pressure of 60 ℃ until the solid content in the final concentrated solution is 3wt%, and carrying out freeze drying treatment to obtain the moringa leaf extract 2.
Analysis of results
1. Chemical composition and particle size analysis
As shown in Table 1, the measurable content of the soluble total sugar in the moringa oleifera leaf dry powder M1 after being crushed by liquid nitrogen is significantly higher than that of the common crushed moringa oleifera leaf dry powder M2, and reaches 21.20% +/-0.62%. As can be seen from fig. 1 and table 2, the particle size of moringa oleifera leaf dry powder M1 after liquid nitrogen pulverization is smaller than that of M2, and the specific surface area is larger. The brittleness of the moringa leaves is increased under the cooling effect of liquid nitrogen, the moringa leaves enter the cavity of the mechanical pulverizer, the moringa leaves, the leaves and the gear ring rotate at high speed through the impeller, and the moringa leaves are subjected to comprehensive effects of repeated impact, collision, shearing, friction and the like, so that the moringa leaves are fully crushed, but common crushing cannot be realized. From fig. 2, it can be seen that when the moringa oleifera leaf powder is observed under an optical microscope at 40 times magnification, the finer powder is more likely to absorb moisture to form finer spherical powder particles, and the distribution in water is more extensive. The moringa leaf raw material is more beneficial to subsequent extraction treatment through higher-degree crushing processing, the dispersibility and the utilization rate of the moringa leaf raw material are improved, the advantages of less raw materials and high yield are realized, and the use of liquid nitrogen for crushing the moringa leaf dry powder M1 is beneficial to the pretreatment utilization and production of the moringa leaves in foods.
TABLE 1 basic chemical composition of Moringa leaf powder
The different letters in table 1 indicate significant differences between groups (p < 0.05); the content is mass percent.
TABLE 2 particle size distribution values of Moringa leaf powder
The different letters in table 2 indicate significant differences between groups (p < 0.05).
2. Color difference analysis of moringa oleifera leaf polysaccharide extract
As can be seen from table 3, the total chromaticity and the total brightness of the moringa oleifera leaf polysaccharide in the extraction process are significantly changed, the color of the polysaccharide extract MEA, MEB, MEC after the enzymatic-high-temperature extraction-fermentation treatment in the examples is the most gentle and bright, the red chromaticity and the yellow chromaticity of the polysaccharide extract ME2 after the enzymatic-high-temperature extraction without fermentation treatment in the comparative examples are significantly reduced (p < 0.05), the total brightness of the water extracts AE1 and AE2 obtained only by the medium-temperature water extraction without enzymolysis wall breaking and the high-temperature extraction is the lowest, and various chromaticities are deep. After post-finishing deep processing treatment, the moringa oleifera leaf polysaccharide extract prepared by the method is more glossy at the color value level and is saturated with Wen Liang, and has the application potential as a nutritional food base material.
TABLE 3 color difference values of Moringa oleifera leaf polysaccharide extract during extraction
The different letters in table 3 indicate significant differences between groups (p < 0.05).
3. Yield and total sugar content of moringa oleifera leaf polysaccharide extract
As can be seen from fig. 3 (the different letters in fig. 3 represent significant differences (p < 0.05)) between groups, the yields of the extracts obtained from the different processes of medium-temperature water extraction, enzymatic extraction and enzymatic-high-temperature extraction of moringa oleifera leaves in the comparative example and the polysaccharide extract MEA, MEB, MEC in the examples were >12%, wherein the yields were calculated as follows:
wherein the mass unit of the moringa oleifera leaf extract is gram, and the mass unit of the moringa oleifera leaf dry powder is gram.
As can be seen from fig. 4 (the different letters in fig. 4 indicate significant differences between groups (p < 0.05), ge=dextrose equivalent.) each processing step of the comparative example has a significant elevating effect on the total sugar content in the extract. Compared with the common crushed moringa dry powder aqueous extract AE2, the liquid nitrogen crushed moringa dry powder aqueous extract AE1 has the advantages that the total sugar content is obviously improved (p < 0.05) under the condition that the yield is not different, the total sugar content of the extract ME1 subjected to enzymolysis wall breaking treatment is obviously improved (p < 0.05) compared with the total sugar content of the aqueous extract AE1, and the total sugar content of the extract ME2 subjected to enzymolysis-high temperature extraction is obviously improved (p < 0.05) compared with the total sugar content of the enzymolysis extract ME1. In the example, the total sugar content of the polysaccharide extract MEA, MEB, MEC is increased from 21.23% to more than 50% compared with the total sugar content of the water extract AE1, the total sugar content is gradually increased to more than 2 times of the original total sugar content, and the extracts in the comparative examples cannot be obtained. The preparation method can effectively improve the total sugar content and purity of the moringa oleifera leaf polysaccharide extract, and has the effect of promoting the extraction efficiency of moringa oleifera leaf polysaccharide components.
4. Analysis of glucose blocking ability in Moringa oleifera leaf polysaccharide extract
As can be seen from fig. 5 (the different letters in fig. 5 indicate significant differences between groups (p < 0.05)), when all samples were at a concentration of 10mg/mL (concentration in the final volume of digestion), the samples had some glucose blocking effect and the rate of glucose diffusion into the dialysis bag's external fluid was gradually reduced during dialysis. In the comparative example, the liquid nitrogen crushed moringa leaves M1 have stronger blocking effect on glucose diffusion than the common crushed moringa leaves M2, so that the blocking effect of the water extract AE1 is stronger than that of AE 2; compared with the water extract, the enzymolysis extract ME1 has more active ingredients and better activity; the enzyme method-high temperature extract ME2 has stronger glucose diffusion blocking activity than the single enzymolysis extract ME1, and the dialysis delay index (p < 0.05) of the moringa oleifera leaf extract to glucose can be obviously improved by both the enzyme method and the high temperature extraction. The enzymatic method-high temperature extraction-yeast fermentation polysaccharide extract MEA, MEB, MEC in the embodiment has the strongest glucose diffusion blocking capability, can further increase the total sugar ratio so as to improve the sugar inhibition activity, and reaches 37.40% +/-1.04% at maximum after fermentation. The moringa oleifera leaf polysaccharide extract obtained by the preparation method can effectively block glucose diffusion.
5. Analysis of cholate adsorption ability and cholesterol micelle dissolution inhibition ability in moringa oleifera leaf polysaccharide extract
As can be seen from fig. 6 (the different letters in fig. 6 indicate that there is a significant difference (p < 0.05)) between groups, and fig. 7 (the different letters in fig. 7 indicate that there is a significant difference (p < 0.05)) between groups, when all the sample concentrations are 5mg/mL (the concentrations in the final volume of digestion), all the samples have a certain adsorption effect on the gastrointestinal cholate during the simulated digestion, have an inhibitory effect on the solubility of cholesterol in the gastrointestinal micelles, and the two lipid lowering activity change trends are consistent. Compared with the dry powder M1 of the moringa oleifera leaves, the water extract AE1 of the moringa oleifera leaves in the comparative example has obviously enhanced lipid absorption inhibition effect (p < 0.05), but in the examples, the absorption rate of cholate and the dissolution inhibition rate of cholesterol of the polysaccharide extract MEA, MEB, MEC of the enzymatic-high-temperature extraction-yeast fermentation are obviously improved (p < 0.05), the maximum lipid absorption inhibition activity is up to more than 50% after fermentation, and the strongest lipid absorption inhibition activity cannot be achieved in the absence of any processing step. The increase of the total sugar content in the polysaccharide extract results in an increase of the viscosity of the sample solution during digestion, an increase of the adsorption capacity for lipid substances and a competitive decrease of the dissolved concentration in the gastrointestinal tract. The moringa oleifera leaf polysaccharide extract obtained by the preparation method has the effect of obviously inhibiting excessive absorption of lipid by gastrointestinal tracts.
The moringa oleifera leaf dry powder M1, the moringa oleifera leaf aqueous extract AE1, the enzymolysis extract ME1 and the enzymolysis-high temperature extract ME2 in the comparative example all have a certain effect of blocking the glucose diffusion and adsorbing lipid substances in the simulated digestion process, and the total sugar content is gradually improved under the condition that the extraction yields are not obviously different, and the metabolic activity of glycolipid is also gradually improved. The total sugar content and the hypoglycemic control activity of the enzymatic-high-temperature extraction-yeast fermentation polysaccharide extract MEA, MEB, MEC in the embodiment can reach the highest, which indicates that the total sugar content of MEA, MEB, MEC is the highest and is more than 50% under the condition of the same quality of the extract; the glucose dialysis delay index of MEA, MEB, MEC is highest at a concentration of 10mg/mL in the final volume of digestion, greater than 37%; the absorption rate of MEA, MEB, MEC for bile acid salt and the dissolution inhibition rate of cholesterol micelle are also highest at a concentration of 5mg/mL in the final volume of digestion, which is more than 50%. The moringa oleifera leaf dry powder belongs to a primary moringa oleifera leaf product, and can be extracted more easily by crushing with liquid nitrogen, and although the moringa oleifera leaf dry powder shows a certain activity of adjusting the absorption of glycolipid, the moringa oleifera leaf dry powder is obviously weaker than the extracted and refined moringa oleifera leaf polysaccharide extract, and the moringa oleifera leaf active substance can have higher release rate and activity after further extraction and processing. The enzymolysis in the embodiment can make the color of the extract brighter and plump, the extraction is more sufficient, the high-temperature extraction can promote the dissolution of components with high efficiency and simultaneously has the effects of sterilization and enzyme deactivation, the sterile condition is provided for inoculating yeast to eliminate small molecular saccharides, the purity of polysaccharide is improved again by consuming a large amount of small molecular saccharides and partial proteins in the yeast fermentation process, the residual small molecular substances can be removed by ethanol precipitation, the condition of the yeast fermentation process is mild and easy to operate, the time consumption is short, the waste is less, and the obvious effect can be achieved by adding a small amount of the small molecular saccharides. The moringa oleifera leaf polysaccharide extract finally obtained in the embodiment has the function of regulating the absorption activity of glycolipid, and has the potential of being used as a food base material to be processed into food and health care products.
The invention provides a moringa oleifera leaf polysaccharide extract with a function of regulating glycolipid absorption, and a preparation method and application thereof. According to the preparation method, liquid nitrogen is adopted for crushing to reduce the particle size of powder, the water-soluble polysaccharide is extracted through enzymatic wall breaking, high-temperature extraction is adopted to dissolve more polysaccharide components, meanwhile, enzyme sterilization can be realized, the purity of the polysaccharide is improved after yeast fermentation treatment, the target product has dual activities of regulating and controlling sugar and lipid absorption, the preparation time is less than 2 days, the extraction time is greatly shortened, the required processing equipment is simple and easy to operate, the food processing specification is met, and the method has potential of industrial popularization and application. The invention has obvious technical advantages and advancement.
The above examples are only preferred embodiments of the present invention, and are intended to be illustrative of the present invention, not limiting, and variations, substitutions, modifications, etc. which would be apparent to those skilled in the art without departing from the spirit of the present invention are to be construed as being within the scope of the present invention.
Claims (9)
1. A preparation method of moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect is characterized by comprising the following steps:
(1) Crushing by liquid nitrogen: crushing the dried moringa leaves into dry powder by a liquid nitrogen crushing device to obtain moringa leaf dry powder;
(2) Enzymatic hydrolysis of cellulase: uniformly mixing the moringa leaf dry powder obtained in the step (1) with water, regulating the pH, adding cellulase after fully and uniformly mixing, and stirring at constant temperature for enzymolysis to obtain a suspension 1;
(3) Heating and extracting: heating and extracting the suspension 1 to obtain a suspension 2;
(4) And (3) yeast fermentation: adding active dry yeast into the suspension 2, and fermenting at constant temperature to obtain a suspension 3; the mass ratio of the added active dry yeast to the water in the step (2) is 1g (200-1000) mL, the fermentation temperature is 26-30 ℃, the fermentation time is 1-3 h, and the rotating speed is 110-130 rpm;
(5) Centrifuging and concentrating: centrifuging the suspension 3, and concentrating the supernatant under reduced pressure to obtain an extract 1;
(6) Ethanol precipitation: adding ethanol solution into the extract 1, fully and uniformly mixing to obtain mixed solution, standing, centrifuging, adding water into the precipitate, heating for fully dissolving, and concentrating under reduced pressure to obtain extract 2;
(7) And (3) freeze drying: and (3) freeze-drying the extracting solution 2 to obtain the moringa oleifera leaf polysaccharide extract.
2. The method for preparing moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect according to claim 1, wherein in the step (1), the condition of liquid nitrogen pulverization is pulverizing at-150 to-120 ℃.
3. The method for preparing moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect according to claim 1, wherein in the step (2), the feed liquid ratio of moringa oleifera leaf dry powder to water is 1:8-1:12 g/mL; adjusting the pH value to 4.0-5.0 by using hydrochloric acid, adding cellulose with the mass of 0.25-0.75% of the moringa oleifera leaf dry powder obtained in the step (1), stirring at the speed of 120-180 r/min, and performing enzymolysis at the temperature of 45-55 ℃ for 1-3 h.
4. The method for preparing moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect according to claim 1, wherein in the step (3), the heating extraction temperature is 90-120 ℃, and the heating extraction time is 20-40 min.
5. The method for preparing moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect according to claim 1, wherein in the step (5), the centrifugation temperature is 4-8 ℃, the centrifugation force is 6000-8000 g, and the centrifugation time is 15-25 min; the temperature of the reduced pressure concentration is 55-65 ℃, and the solid content in the extracting solution 1 is 4-10 wt%.
6. The method for preparing moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect according to claim 1, wherein in the step (6), the volume of the added ethanol solution accounts for 70% -90% of the total volume of the mixed solution, the standing temperature is 4-8 ℃, the standing time is 2-4 h, the centrifugation temperature is 4-8 ℃, the centrifugal force is 4000-8000 g, and the centrifugation time is 15-25 min.
7. The method for preparing moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect according to claim 1, wherein in the step (6), water is added in an amount which is 100-150 times the mass of the precipitate, the heating and dissolving temperature is 50-60 ℃, the heating and dissolving time is 1-3 hours, the decompressing and concentrating temperature is 55-65 ℃, and the solid content in the extract 2 is 2-4 wt%.
8. The method of any one of claims 1-7, wherein a moringa oleifera leaf polysaccharide extract with glycolipid absorption regulating effect is obtained.
9. The use of a moringa oleifera leaf polysaccharide extract with a function of regulating the absorption of glycolipid as claimed in claim 8 for preparing a hypoglycemic and lipid-controlling food, a health product or a medicament.
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