CN114521650B - Jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics promoter and preparation and application thereof - Google Patents
Jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics promoter and preparation and application thereof Download PDFInfo
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Classifications
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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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/045—Organic compounds containing nitrogen as heteroatom
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- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/72—Rhamnaceae (Buckthorn family), e.g. buckthorn, chewstick or umbrella-tree
- A61K36/725—Ziziphus, e.g. jujube
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/736—Prunus, e.g. plum, cherry, peach, apricot or almond
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- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
The invention belongs to the technical field of probiotic accelerators, and discloses a jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal tract probiotic accelerator, and preparation and application thereof. The method comprises the following steps: 1) Extracting pigment in fructus Jujubae and semen Armeniacae amarum with ethanol water solution, filtering, and drying the filtrate to obtain fructus Jujubae and semen Armeniacae amarum pigment; 2) Extracting the filtered residue in buffer solution with pH of 7-10 to obtain a mixture of polysaccharide and protein; 3) Reacting the mixture of polysaccharide and protein to obtain a jujube apricot glycoprotein conjugate; 4) Preparing the jujube apricot pigment and the jujube apricot glycoprotein conjugate into solutions respectively, mixing the jujube apricot pigment solution and the jujube apricot glycoprotein conjugate solution under the condition of stirring, and passing through a microporous filter membrane to obtain the jujube apricot glycoprotein-entrapped jujube pigment nano intestinal probiotics accelerant. The accelerator can inhibit escherichia coli and promote the remarkable proliferation of intestinal lactobacillus and bifidobacterium.
Description
Technical Field
The invention belongs to the technical field of probiotic accelerators, and particularly relates to a jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal tract probiotic accelerator, and a preparation method and application thereof.
Background
Intestinal flora is a normal microorganism in the intestinal tract of the host and is of a wide variety. Generally, a healthy adult has about 10 microorganisms in the intestinal tract 14 And about 10 times of the number of human cells, can form a complex and unique system in vivo to regulate the normal physiological functions of a host, and the composition, metabolism and the like of the flora can be influenced by the surrounding environment of the host. Wherein the intestinal probiotics comprise lactobacillus and bifidobacterium, etc., which are essential elements for human health, and can synthesize various vitamins to participate in food digestionPromoting intestinal peristalsis, inhibiting growth of pathogenic bacteria, decomposing harmful and toxic substances, etc. By regulating intestinal flora, increasing probiotics, the preparation can inhibit intestinal diseases caused by various pathogenic bacteria, and can regulate metabolism and immune function of human body.
Fructus Jujubae and semen Armeniacae amarum have effects of regulating gastrointestinal function and loosening bowel to relieve constipation, and CN111616353A and CN105285984A both disclose a formula for regulating intestinal flora, comprising fructus Jujubae or its extract; CN104905352a and CN108208504a respectively disclose a preparation method of a health beverage, which comprises almond raw materials, and has the effect of regulating intestinal flora, however, the formulations adopt jujube and almond raw materials or aqueous extracts thereof through oral administration, and as the jujube and the almond contain some functional components which are unstable and are easily destroyed in gastrointestinal tract environment, the purpose of inhibiting intestinal harmful bacteria and promoting the proliferation of probiotics can not be achieved through oral administration.
Disclosure of Invention
In order to overcome the defects and shortcomings in the prior art, the primary purpose of the invention is to provide a jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant and a preparation method thereof.
The invention further aims to provide an application of the jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant in preparing intestinal flora regulating drugs.
The aim of the invention is achieved by the following technical scheme.
A nanometer intestinal probiotics promoter containing fructus Jujubae and fructus Pruni glycoprotein is prepared from fructus Jujubae polysaccharide and semen Armeniacae amarum protein conjugate as shell and fructus Jujubae and fructus Pruni pigment as core.
The preparation method of the jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant comprises the following steps:
1) Extracting pigment in fructus Jujubae and semen Armeniacae amarum with ethanol water solution, filtering, and drying the filtrate to obtain fructus Jujubae and semen Armeniacae amarum pigment; the mass ratio of the jujube to the almond in the step 1) is 1: (0.5 to 3), preferably 1: (0.9-3); the ethanol aqueous solution is ethanol aqueous solution with volume fraction of 60-80%; the mass ratio of the total mass of the Chinese dates and the almonds to the ethanol aqueous solution is 1: (8-20), wherein the extraction condition is that leaching is carried out at 60-80 ℃ for 2-5h; the drying is vacuum drying (50-60 ℃ C., 0.01-0.1MPa drying for 3-5 h); before extraction, crushing the Chinese date and the almond, and sieving with a 10-30 mesh sieve;
2) Extracting the filtered filter residues in a buffer solution with the pH value of 7-10 to obtain a mixture of polysaccharide and protein; in the step (2), the mass ratio of the chestnut to the almond is 1: (0.5-3), preferably 1: (0.8-3); the mass ratio of the filter residue to the buffer solution is 1: (10-30); filtering, dialyzing, and freeze-drying to obtain polysaccharide and protein mixture; the dialysis refers to dialysis by a membrane with the molecular weight cutoff of 50-200kDa, the dialysis time is 24-48h, the freeze drying refers to freeze drying the dialysed trapped fluid, and the freeze drying time is 24-48h; the freeze-drying temperature is-10 to-50 ℃; extracting for 1-3h at 50-60 ℃;
3) Reacting the polysaccharide and protein mixture at 50-70deg.C and relative humidity of 65-85% for 24-72 hr to obtain fructus Jujubae and apricot glycoprotein conjugate;
4) Preparing the jujube apricot pigment into a solution by adopting an ethanol water solution with the volume fraction of 60-80% to obtain a jujube apricot pigment solution; preparing the jujube apricot glycoprotein conjugate into a solution by adopting a buffer solution with the pH of 7-10 to obtain a jujube apricot glycoprotein conjugate solution; mixing the jujube apricot pigment solution and the jujube apricot glycoprotein conjugate solution under the condition of stirring, and filtering the mixture through a microporous filter membrane to obtain the jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant.
The mass ratio of the jujube apricot pigment to the jujube apricot glycoprotein conjugate in the step 4) is 1: (2-20), preferably 1: (5-20).
The mass concentration of the jujube apricot pigment solution in the step (4) is 5-10%, and the mass concentration of the jujube apricot glycoprotein conjugate solution is 5-10%.
The stirring rotating speed in the step (4) is 5000-20000 revolutions/min; the mixing is stirring mixing, and the mixing time is 10-60min; the mixing means that the jujube apricot pigment solution is added into the jujube apricot glycoprotein conjugate solution, and the mixture is stirred and mixed.
The microporous filter membrane in the step (4) is a microporous filter membrane with the thickness of 0.1-0.5 mu m;
the buffer solution in the step (2) and the buffer solution in the step (4) are respectively one of phosphate buffer solution and Tris-HCl buffer solution.
The application of the jujube apricot glycoprotein-entrapped jujube apricot pigment nano intestinal probiotics accelerant in preparing intestinal flora regulating products (such as medicines and health-care foods) can inhibit escherichia coli after oral administration so as to promote the remarkable proliferation of lactobacillus and bifidobacterium in the intestinal tract, thereby preventing diseases caused by harmful bacteria in the intestinal tract.
The principle of the invention is as follows: extracting pigment from fructus Jujubae and fructus Pruni with ethanol, extracting with buffer solution, dialyzing, separating to obtain fructus Jujubae polysaccharide (composed of arabinose-galactose-glucose-rhamnose, with terminal reducibility) and semen Armeniacae amarum protein mixture, and connecting fructus Jujubae polysaccharide and semen Armeniacae amarum protein to form fructus Jujubae and fructus Pruni glycoprotein conjugate. After the jujube apricot glycoprotein and the jujube apricot glycoprotein conjugate are subjected to nanoemulsification, the jujube apricot glycoprotein conjugate has amphipathy, the hydrophilic glycoprotein is outside, the hydrophobic group is connected with pigment molecules inside, and the nano structure with the jujube apricot glycoprotein as a shell and the jujube apricot glycoprotein as a core is formed. The jujube apricot glycoprotein is acid glycoprotein, is stable in gastric juice, is hydrolyzed in intestinal tracts to release pigment, the glycoprotein can strengthen the adhesion with bacteria, and the jujube apricot glycoprotein is a polyphenol compound, can inhibit harmful bacteria, thereby promoting the proliferation of intestinal probiotics and keeping the health of human bodies.
Compared with the prior art, the invention has the following advantages and effects:
(1) The invention prepares the jujube apricot glycoprotein-coated jujube apricot pigment nano-preparation, which keeps the stability of the jujube apricot pigment in gastric juice.
(2) The invention overcomes the defects that jujube polysaccharide or almond protein can only provide single nutrition and can not inhibit harmful bacteria, and can simultaneously realize the proliferation of beneficial bacteria in intestinal tracts and the inhibition of harmful bacteria.
(3) The invention has mild reaction condition and is easy for industrial production.
Detailed Description
The present invention will be described in further detail with reference to examples, but embodiments of the present invention are not limited thereto.
Example 1
(1) The mass ratio of the jujube to the almond is 1:1 crushing, sieving with a 10-mesh sieve, adding 80% ethanol water solution with the volume fraction of 8 times of the mass of the mixture, leaching for 5 hours at 60 ℃, filtering, and drying the filtrate at 50 ℃ under 0.01MPa for 3 hours to obtain jujube apricot pigment;
(2) Extracting the residue with 10 times of 50 deg.C pH7 phosphate buffer solution for 3 hr, filtering, dialyzing the filtrate with 50kDa membrane for 48 hr, and freeze-drying the retentate (freeze-drying temperature is-50deg.C) for 24 hr to obtain polysaccharide and protein mixture;
(3) Reacting the jujube apricot polysaccharide and protein mixture at 50 ℃ with relative humidity of 65% for 72 hours to obtain the jujube apricot glycoprotein conjugate.
(4) Preparing 10g of jujube apricot pigment into a 5% solution by using a 70% ethanol water solution; 50g of the jujube apricot glycoprotein conjugate is prepared into a 5% solution by using a pH7 phosphate buffer; adding the jujube apricot pigment solution into the jujube apricot glycoprotein conjugate solution under the stirring of 5000 revolutions per minute, and stirring for 10 minutes, and then passing through a 0.1 mu m microporous filter membrane to obtain the jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant.
Example 2
(1) The mass ratio of the jujube to the almond is 1:3 crushing, sieving with a 30-mesh sieve, adding 60% ethanol water solution with the volume fraction of 20 times of the mass of the mixture, leaching for 2 hours at 80 ℃, filtering, and drying filtrate at 60 ℃ under 0.1MPa for 5 hours to obtain jujube apricot pigment;
(2) Extracting the filter residue with 30 times of phosphate buffer solution with pH10 at 60deg.C for 1 hr, filtering, dialyzing the filtrate with 200kDa membrane for 24 hr, and lyophilizing the retentate for 48 hr to obtain polysaccharide and protein mixture;
(3) And (3) reacting the jujube apricot polysaccharide and protein mixture for 24 hours at 70 ℃ with the relative humidity of 85% to obtain the jujube apricot glycoprotein conjugate.
(4) Preparing 10g of jujube apricot pigment into a solution with the volume fraction of 80% of ethanol water solution of 10 wt%; 200g of the jujube apricot glycoprotein conjugate is prepared into a 10wt% solution by using a pH10 phosphate buffer; adding the jujube apricot pigment solution into the jujube apricot glycoprotein conjugate solution under stirring at 20000 rpm, and stirring for 60min, and then passing through a 0.5 μm microporous filter membrane to obtain the jujube apricot glycoprotein-entrapped jujube pigment nano intestinal probiotics accelerant.
Example 3
(1) The mass ratio of the jujube to the almond is 1:2 crushing, sieving with a 20-mesh sieve, adding 70% ethanol water solution with the volume fraction of 10 times of the mass of the mixture, leaching for 3 hours at 70 ℃, filtering, and drying the filtrate at 55 ℃ under 0.05MPa for 4 hours to obtain jujube apricot pigment;
(2) Adding 20 times of 55 ℃ pH8Tris-HCl buffer solution into the filter residue, extracting for 2 hours, filtering, dialyzing the filtrate for 36 hours by using a membrane with a molecular weight cut-off of 100kDa, and freeze-drying the cut-off for 36 hours to obtain a jujube apricot polysaccharide and protein mixture;
(3) Reacting the jujube apricot polysaccharide and protein mixture at 60 ℃ with relative humidity of 75% for 48 hours to obtain a jujube apricot glycoprotein conjugate;
(4) Preparing 10g of jujube apricot pigment into 8% ethanol water solution with the volume fraction of 75%; preparing 100g of jujube apricot glycoprotein conjugate into 8% solution by using pH8Tris-HCl buffer solution; adding the jujube apricot pigment solution into the jujube apricot glycoprotein conjugate solution under 10000 revolutions per minute of stirring, and stirring for 40 minutes, and then passing through a 0.3 mu m microporous filter membrane to obtain the jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant.
Example 4
(1) The mass ratio of the jujube to the almond is 1:1.5 crushing, sieving with a 15-mesh sieve, adding a 65% ethanol water solution with the volume fraction of 12 times of the mass of the mixture, leaching at 75 ℃ for 3 hours, filtering, and drying the filtrate at 50 ℃ under 0.03MPa for 3.5 hours to obtain jujube apricot pigment;
(2) Extracting the filter residue with 15 times of 50 deg.C pH9 phosphate buffer solution for 1.5 hr, filtering, dialyzing the filtrate with 80kDa membrane for 40 hr, and freeze-drying the retentate for 30 hr to obtain fructus Jujubae polysaccharide and protein mixture;
(3) Reacting the jujube apricot polysaccharide and protein mixture at 55 ℃ with the relative humidity of 70% for 40 hours to obtain a jujube apricot glycoprotein conjugate;
(4) Preparing 10g of jujube apricot pigment into a 6% solution by using a 60% ethanol water solution with volume fraction; 80g of the jujube apricot glycoprotein conjugate is prepared into a 6% solution by using a pH9 phosphate buffer; adding the jujube apricot pigment solution into the jujube apricot glycoprotein conjugate solution under 6000 rpm stirring, and stirring for 30min, and then passing through a 0.2 mu m microporous membrane to obtain the jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant.
Example 5
(1) The mass ratio of the jujube to the almond is 1:2.5 crushing, sieving with a 25-mesh sieve, adding a 75% ethanol water solution with the volume fraction of 16 times of the mass of the mixture, leaching for 4 hours at 65 ℃, filtering, and drying the filtrate at 60 ℃ under 0.06MPa for 4.5 hours to obtain jujube apricot pigment;
(2) Adding 25 times of 60 ℃ Tris-HCl buffer solution with pH of 7.5 into the filter residue, extracting for 2.5 hours, filtering, dialyzing the filtrate for 30 hours by using a membrane with a molecular weight cutoff of 150kDa, and freeze-drying the trapped liquid for 40 hours to obtain a jujube apricot polysaccharide and protein mixture;
(3) Reacting the jujube apricot polysaccharide and protein mixture for 30 hours at 65 ℃ with the relative humidity of 80% to obtain a jujube apricot glycoprotein conjugate;
(4) Preparing 10g of jujube apricot pigment into 8% solution by using 65% ethanol water solution; 160g of the jujube apricot glycoprotein conjugate is prepared into 8% solution by using a pH7.5Tris-HCl buffer solution; adding the jujube apricot pigment solution into the jujube apricot glycoprotein conjugate solution under 12000 r/min stirring, stirring for 25min, and passing through a 0.4 μm microporous filter membrane to obtain the jujube apricot glycoprotein-entrapped jujube pigment nano intestinal probiotics accelerant.
Comparative example 1
The preparation method of example 1 was followed, but step (3) was omitted, and the jujube apricot glycoprotein conjugate of step (4) was replaced with the jujube apricot polysaccharide and protein mixture of step (2), and the product was control 1.
Comparative example 2
The preparation was as in example 1, but in step (4) the jujube apricot glycoprotein conjugate was replaced with lecithin, and the product was obtained as control 2.
Test 1 analysis of compositions of the jujube apricot glycoprotein conjugates prepared in examples 1-5
The method comprises the following steps: the jujube apricot glycoprotein conjugate prepared in the examples 1-5 is taken to measure the sugar content in a sample according to the agricultural industry standard of the people's republic of China, the measurement of the content of crude polysaccharide in edible fungi (NY/T1676-2008), and the protein content in the sample is measured according to the Coomassie Brilliant blue method (SN/T3926-2006) according to the inspection and quarantine industry standard of the people's republic of China, the measurement of the protein content in milk, egg and bean foods.
Results: the sugar and protein contents of the jujube apricot glycoprotein conjugates prepared in examples 1-5 are shown in Table 1, and each 100g of the conjugates contains 25.8-34.2g of sugar and 63.5-71.4g of protein, which indicates that the conjugates mainly comprise sugar and protein.
TABLE 1 content composition of jujube apricot glycoprotein conjugate
| Sample source | Sugar content% | Protein content% |
| Example 1 | 31.3 | 63.5 |
| Example 2 | 27.7 | 70.2 |
| Example 3 | 34.2 | 64.6 |
| Example 4 | 25.8 | 71.4 |
| Example 5 | 28.1 | 69.3 |
Test 2 determination of the nanometer particle size and the drug-loading amount of the jujube apricot pigment in the jujube apricot glycoprotein-coated jujube apricot pigment nanometer intestinal tract probiotic accelerant prepared in examples 1-5
The method comprises the following steps: the jujube apricot glycoprotein-entrapped jujube apricot pigment nano preparation prepared in examples 1-5, comparative example 1 and comparative example 2 are subjected to particle size measurement by a Markov nanometer particle sizer, and the jujube apricot pigment is removed by a microcolumn centrifugation method in a proper amount because the jujube apricot pigment belongs to anthocyanin compounds, the content of the jujube apricot pigment is measured by a measuring-ultraviolet/visible spectrophotometry method of procyanidine in DB 12/T885-2019 plant extracts, and the drug-loading amount is calculated.
Results: the particle size and the content of the jujube glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant prepared in the embodiment 1-5 are shown in the table 2, the particle size is 216-284nm, the drug loading rate is 4.8-10.4%, and the product is a nano preparation loaded with the jujube apricot pigment. Comparative example 1 was drug-free, indicating that jujube polysaccharide and almond protein did not possess entrapment capacity; control 2 has a small amount of drug loading, which means that the liposome can be coated with jujube apricot pigment, but the drug loading is low.
Table 2 particle size and drug-loading rate of nano intestinal probiotics promoter of jujube and apricot pigment coated with glycoprotein of jujube and apricot
| Sample source | Average particle diameter nm | Jujube apricot pigment drug-loading rate% |
| Example 1 | 284 | 10.4 |
| Example 2 | 216 | 4.8 |
| Example 3 | 253 | 7.5 |
| Example 4 | 278 | 8.6 |
| Example 5 | 236 | 5.6 |
| Comparative example 1 | 155 | 0 |
| Comparative example 2 | 248 | 1.2 |
Test 3 stability determination of jujube apricot glycoprotein-coated jujube apricot pigment nanometer intestinal probiotic promoter prepared in examples 1-5 in simulated gastrointestinal fluids
The method comprises the following steps: stability evaluation was performed on jujube apricot glycoprotein-entrapped jujube apricot pigment nano-formulations using simulated artificial gastric fluid (containing 1% pepsin, ph=1.2) and simulated artificial intestinal fluid (containing 1% trypsin, ph=6.8). 1mL of the jujube apricot glycoprotein-coated apricot pigment nano preparation prepared in examples 1-5 and comparative example 2 are added into 4mL of simulated artificial gastric fluid or intestinal fluid, the mixture is subjected to shaking incubation at 37 ℃ and 100rpm, 200 mu L of the mixture is sampled every 2h time intervals, exuded free pigment is removed by a microcolumn centrifugation method, and the obtained filtrate is transferred into a 5mL volumetric flask, subjected to methanol constant volume and subjected to ultrasonic demulsification. Measuring procyanidine content in DB 12/T885-2019 plant extract by ultraviolet/visible spectrophotometry, measuring fructus Jujubae and fructus Pruni pigment content, and calculating the residual pigment percentage of the preparation at each time point with 0 time drug concentration of 100%.
Results: the nano intestinal probiotics promoter of the jujube apricot glycoprotein-coated jujube apricot pigment prepared in the examples 1-5 still keeps more than 80% of the pigment in simulated gastric fluid for 4 hours, but is degraded more quickly in intestinal fluid, and the results are shown in Table 3. The product has the effect of improving gastric juice stability, and can rapidly release pigment in intestinal tract. Comparative example 2 showed fast release in gastrointestinal fluids, indicating that the liposomes were unstable in gastrointestinal fluids.
TABLE 3 retention of pigment in simulated gastrointestinal fluids (%)
Test 4 effects of the jujube apricot glycoprotein-coated jujube apricot pigment nanometer intestinal probiotic promoter prepared in example 1
The method comprises the following steps: 35 male SD rats of 250+ -25 g were randomly divided into 7 groups of 5. The normal rat feed is given in group 1, the high-fat feed (87.6% standard feed, 2% cholesterol, 0.2% sodium cholate and 10% lard) is given in group 2, and the high-fat feed is respectively given in groups 3-7, and then the mixture of jujube apricot polysaccharide and protein, jujube apricot pigment, jujube apricot glycoprotein conjugate, jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant and control example 2 prepared in example 1 are respectively infused in stomach, and the dosage is 1g/100g of body weight twice a day. Rat faeces were taken at 15 and 30 days, respectively, and subjected to 16sRNA metagenomic sequencing of intestinal flora and probiotics (e.g.bifidobacteria and lactobacilli), and abundance analysis (relative percentages) of the various probiotics and Escherichia coli.
Results: the results are shown in Table 4, which shows that the probiotics of the group 2 model rats are obviously reduced compared with the normal group, but the escherichia coli is increased, and the groups 3-4 and 7 are improved compared with the second group, but the differences from the normal group are not great; the group 5 is obviously increased, and the group 6 is optimal, so that the jujube apricot glycoprotein conjugate can provide comprehensive nutrition for intestinal probiotics and promote the proliferation of the intestinal probiotics, and the jujube glycoprotein-entrapped jujube apricot pigment nano intestinal probiotics accelerant not only can provide multiple nutrition, but also can better promote the proliferation of the intestinal probiotics through the inhibition of the jujube apricot pigment to harmful flora escherichia coli.
TABLE 4 relative abundance of probiotics in rat feces (%)
The above examples of the present invention are merely illustrative of the present invention and are not intended to limit the embodiments of the present invention. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. Any modification, equivalent replacement, improvement, etc. which come within the spirit and principles of the invention are desired to be protected by the following claims.
Claims (5)
1. A preparation method of a jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant is characterized by comprising the following steps of: the method comprises the following steps:
1) Extracting pigment in fructus Jujubae and semen Armeniacae amarum with ethanol water solution, filtering, and drying the filtrate to obtain fructus Jujubae and semen Armeniacae amarum pigment;
2) Extracting the filtered residue in buffer solution with pH of 7-10 to obtain a mixture of polysaccharide and protein;
3) Reacting the mixture of polysaccharide and protein at 50-70deg.C and relative humidity of 65-85% for 24-72 hr to obtain fructus Jujubae and apricot glycoprotein conjugate;
4) Preparing the jujube apricot pigment into a solution by adopting an ethanol water solution with the volume fraction of 60-80% to obtain a jujube apricot pigment solution; preparing the jujube apricot glycoprotein conjugate into a solution by adopting a buffer solution with the pH of 7-10 to obtain a jujube apricot glycoprotein conjugate solution; mixing the jujube apricot pigment solution with the jujube apricot glycoprotein conjugate solution under the condition of stirring, and passing through a microporous filter membrane to obtain a jujube apricot glycoprotein-coated jujube apricot pigment nano intestinal probiotics accelerant;
the mass ratio of the jujube to the almond in the step 1) is 1: (0.9-3); the ethanol water solution in the step 1) is ethanol water solution with volume fraction of 60-80%; the mass ratio of the total mass of the Chinese dates and the almonds to the ethanol aqueous solution in the step 1) is 1: (8-20), wherein the extraction condition in the step 1) is that the extraction is carried out at 60-80 ℃ for 2-5h;
the buffer solution in the steps 2) and 4) is phosphate buffer solution; or the buffer solution in the steps 2) and 4) is Tris-HCl buffer solution;
after the extraction in the step 2), filtering, dialyzing, and freeze-drying to obtain a polysaccharide and protein mixture; the dialysis in step 2) refers to dialysis with a membrane having a molecular weight cut-off of 50-200 kDa;
the mixing in the step 4) is stirring mixing, and the mixing time is 10-60min;
the microporous filter membrane in the step 4) is a microporous filter membrane with the diameter of 0.1-0.5 mu m;
the mass ratio of the filter residue to the buffer solution in the step 2) is 1: (10-30);
the mass ratio of the jujube apricot pigment to the jujube apricot glycoprotein conjugate in the step 4) is 1: (5-20); the stirring rotation speed in the step 4) is 5000-20000 revolutions/min;
the mass concentration of the jujube apricot pigment solution in the step 4) is 5-10%, and the mass concentration of the jujube apricot glycoprotein conjugate solution is 5-10%.
2. The method for preparing the jujube apricot glycoprotein-entrapped jujube apricot pigment nano intestinal probiotics accelerant according to claim 1, which is characterized by comprising the following steps: the dialysis time in the step 2) is 24-48 hours, and the freeze drying refers to freeze drying the retentate of the dialysis, wherein the freeze drying time is 24-48 hours; the temperature of freeze drying is-10 to-50 ℃.
3. The method for preparing the jujube apricot glycoprotein-entrapped jujube apricot pigment nano intestinal probiotics accelerant according to claim 1, which is characterized by comprising the following steps: the mixing in the step 4) means that the jujube apricot pigment solution is added into the jujube apricot glycoprotein conjugate solution, and the mixture is stirred and mixed;
the drying in the step 1) is vacuum drying; before extraction in the step 1), the Chinese date and the almond are crushed and sieved by a 10-30-mesh sieve.
4. A nano intestinal probiotics promoter of jujube apricot glycoprotein-entrapped jujube apricot pigment obtained by the preparation method of any one of claims 1-3, which is characterized in that: is a nano preparation which takes the combination of jujube polysaccharide and almond protein as a shell and jujube apricot pigment as a core.
5. The application of the jujube apricot glycoprotein-entrapped jujube and apricot pigment nano intestinal probiotics accelerant in preparing intestinal flora regulating drugs, which is characterized in that: inhibit Escherichia coli and promote proliferation of intestinal lactobacillus and bifidobacterium.
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