METHOD FOR PREPARING A FEEDING INSTANT ASTRAGALUS POLYSACCHARIDE POWDER AND APPLICATION THEREOF
20240058410 ยท 2024-02-22
Assignee
Inventors
Cpc classification
Y02A40/70
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
B01D69/02
PERFORMING OPERATIONS; TRANSPORTING
B01D2311/04
PERFORMING OPERATIONS; TRANSPORTING
A61K2236/39
HUMAN NECESSITIES
A61K2236/331
HUMAN NECESSITIES
A61K2236/37
HUMAN NECESSITIES
Y02P60/87
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
B01D2311/08
PERFORMING OPERATIONS; TRANSPORTING
A23K10/30
HUMAN NECESSITIES
A61K9/14
HUMAN NECESSITIES
A61K2236/53
HUMAN NECESSITIES
International classification
A23K10/30
HUMAN NECESSITIES
B01D61/14
PERFORMING OPERATIONS; TRANSPORTING
B01D69/02
PERFORMING OPERATIONS; TRANSPORTING
Abstract
The present application is related to a method for preparing a feeding instant Astragalus polysaccharide powder and an application thereof. The method includes the following steps: adding deionized water at 65 C. to a dreg of Astragalus membranaceus by water extraction and alcohol precipitation, controlling an alcohol volume fraction at 20%, stirring fully to dissolve the dreg, and after a dreg solution cools to room temperature, removing insoluble thermo-sensitive macromolecular proteins, polysaccharides and solid residues by centrifugation at 3500 revolutions per minute to obtain a clear supernatant. Ultrafiltration treatment is performed by hollow fiber membranes on the supernatant obtained after the centrifugation, and 10-100 kDa polysaccharide components are collected. A low-molecular-weight instant Astragalus polysaccharide powder with high immunocompetence is obtained through an interlayer cold air cooling spray drying process, and the content of active polysaccharide is more than 87%.
Claims
1. A method for preparing a feeding instant Astragalus polysaccharide powder, the method comprising the following steps: 1) adding deionized water at 65 C. to a dreg of Astragalus membranaceus by water extraction and alcohol precipitation, and stirring until the dreg is completely dissolved to prepare a dreg solution; 2) after the dreg solution cools to room temperature, removing insoluble thermo-sensitive macromolecular proteins, polysaccharides and solid residues by centrifugation to obtain a clear supernatant; 3) adding twice mass of deionized water to the clear supernatant, stirring fully, and then performing a secondary ultrafiltration treatment by hollow fiber membranes with pore sizes of 100 KDa and 10 kDa in turn, and collecting 10-100 kDa polysaccharide components; and 4) concentrating the polysaccharide components with a molecular weight between 10-100 kDa collected in the step 3) to 40% of solid contents of the polysaccharide components under vacuum and low temperature, and spray drying to obtain the instant Astragalus polysaccharide powder.
2. The method for preparing the feeding instant Astragalus polysaccharide powder of claim 1, wherein in the step 1), an alcohol volume fraction of the dreg aqueous solution is controlled at 20%.
3. The method for preparing the feeding instant Astragalus polysaccharide powder of claim 1, wherein in the step 1), the dreg of Astragalus membranaceus by water extraction and alcohol precipitation is a semi-solid residue obtained by extracting traditional Chinese medicine Astragalus membranaceus pieces with hot water, removing a solid residue, and adding 3-4 times volume of anhydrous ethanol.
4. The method for preparing the feeding instant Astragalus polysaccharide powder of claim 1, wherein in the step 2), a speed of the centrifugation is 3500 revolutions per minute.
5. The method for preparing the feeding instant Astragalus polysaccharide powder of claim 1, wherein in the step 4), conditions of the spray drying are as follows: an atomizer frequency is 70 Hz, an inlet air temperature is controlled at 140 C., an outlet air temperature is controlled below 60 C., a drying tower interlayer is cooled by cold air, and a bottom of a drying tower is vibrated by a vibrating hammer every 5 minutes to prevent a wall and the bottom of the drying tower from powder deposition and liquefaction foaming.
6. An application of the feeding instant Astragalus polysaccharide powder prepared by the method according to claim 1 in a feed additive.
7. An application of the feeding instant Astragalus polysaccharide powder prepared by the method according to claim 2 in a feed additive.
8. An application of the feeding instant Astragalus polysaccharide powder prepared by the method according to claim 3 in a feed additive.
9. An application of the feeding instant Astragalus polysaccharide powder prepared by the method according to claim 4 in a feed additive.
10. An application of the feeding instant Astragalus polysaccharide powder prepared by the method according to claim 5 in a feed additive.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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DETAILED DESCRIPTION OF THE EMBODIMENTS
[0028] The following provides a further detailed description of the present invention in conjunction with specific embodiments, in order to better understand the technical solutions.
Embodiment 1
[0029] Deionized water that had been heated to 65 C. was added to 50 kilograms of a dreg of Astragalus membranaceus by water extraction and alcohol precipitation from a production enterprise of Astragalus membranaceus injection, the amount of water added was controlled with a precision alcohol meter to control a final alcohol volume fraction at 20%, and stirring lasted for 20 minutes until the dreg of Astragalus membranaceus by water extraction and alcohol precipitation is evenly dissolved. After a dreg solution cooled to room temperature, insoluble residues were removed by centrifugation at 3500 revolutions per minute to obtain a clear supernatant; twice mass of deionized water was added to the supernatant and stirred evenly, and then ultrafiltration was performed by an ultrafiltration membrane with the molecular weight cut-off of 100 KDa, and liquid components below 100 kDa were collected. Then 10-100 kDa polysaccharide components were collected by ultrafiltration treatment with a 10 kDa hollow fiber membrane. Further, through evaporation and concentration under vacuum and low temperature, a concentrated trapped solution with a solid content of 40% was obtained. A spray dryer was used for drying, an atomizer frequency was 70 Hz, inlet air temperature was controlled at 140 C., outlet air temperature was selected at 60 C., a feeding speed was 15 kilograms per hour, a drying tower interlayer was cooled by cold air below 20 C., a bottom of a drying tower was hammered with a hammer every 5 minutes to prevent powder deposition on a wall of the drying tower and liquefaction foaming of high-temperature active polysaccharides, and a dried powder was collected in a workshop equipped with dehumidification equipment to obtain highly a bioactive Astragalus polysaccharide powder. Its appearance was light yellow, easy to absorb moisture, and highly soluble in water, and had no bad odor. A powder yield of spray drying was about 90%, a content of crude polysaccharide was 87%, a drying weight loss was 6%, solvent residues were not detected, a content of heavy metals (arsenic+lead+cadmium+mercury) was <10.0 ppm, and pathogenic bacteria such as coliform bacteria and salmonella were not detected. The highly active low-molecular-weight Astragalus polysaccharide powder has extremely high hygroscopicity, and should be sealed and stored after collection.
Experimental Example 1: Composition Analysis of a Dreg of Astragalus membranaceus by Water Extraction and Alcohol Precipitation and Selection of a Spray Drying Process
[0030] Analysis of main components of the dreg of Astragalus membranaceus by water extraction and alcohol precipitation: the dreg of Astragalus membranaceus by water extraction and alcohol precipitation is the residual waste after water-soluble small-molecule components such as astragaloside IV are extracted from Astragalus membranaceus using the water extraction and alcohol precipitation method. Due to the semi-solid paste-like nature and high viscosity of the dreg, as shown in
TABLE-US-00001 TABLE 1 Composition analysis of a dreg of Astragalus membranaceus by water extraction and alcohol precipitation Dry matter/% Drying Crude Crude Free Reducing Ash loss/% polysaccharide protein protein sugar content 36.15 72.32 13.14 1.15 2.08 2.62
[0031] Grading of polysaccharide alcohol precipitation in the dreg of Astragalus membranaceus by water extraction and alcohol precipitation: the dreg of Astragalus membranaceus by water extraction and alcohol precipitation was dissolved in water with an ethanol concentration below 1%, a supernatant was collected by centrifugation, ethanol was added to the supernatant to a concentration of 10%, a precipitate was collected, then the ethanol concentration of the supernatant was adjusted to 20%, a precipitate was collected, the ethanol concentration of the supernatant was adjusted to 30% and so on, until the ethanol concentration was 90%. The alcohol precipitate was dried by freezing, and the proportion of polysaccharides by alcohol precipitation in each part was analyzed.
[0032] Molecular weight distribution in a 20% alcohol precipitate of a dreg of Astragalus membranaceus by water extraction and alcohol precipitation: the dreg of Astragalus membranaceus by water extraction and alcohol precipitation was dissolved in deionized water until the alcohol content was 20%. Centrifugation was performed to collect precipitates, the precipitates were then added with deionized water for complete dissolution, a supernatant was collected by centrifugation, the supernatant is then passed through a hollow fiber membrane with a molecular weight of 100 KDa to collect a trapped solution, and the polysaccharide content in the supernatant and the trapped solution was measured separately, and the ratio between the two was calculated.
[0033] Influence of air inlet temperature on spray drying of Astragalus polysaccharides: the 10-100 kDa components of Astragalus polysaccharides were adjusted to about 40% of solid contents. At this concentration, it had good fluidity and high solid contents, which was conducive to improving the efficiency of spray drying. Air outlet temperature was controlled below 60 C., an atomizer frequency was 70 Hz, and four temperature gradients including 140, 160, 180, 200 C. were selected. The results are shown in Table 2.
TABLE-US-00002 TABLE 2 Effects of air inlet temperature on spray drying of astragalus polysaccharides Inlet air temperature/ Powder Water C. yield/% content/% Sensory effects 140 89.52 6.0 Slight wall sticking, good spray effect, fine and even powder particles, loose, slightly caking, and light yellow 160 70.17 5.3 Foaming and wall sticking, general spray effect, fine and even powder particles, loose, caking, and light brown yellow 180 48.18 4.5 Severe foaming and wall sticking, good spray effect, caking, and brown yellow 200 30.71 3.8 Severe foaming and wall sticking, difficult to obtain dry powder, large caking, and brown yellow
[0034] Table 2 showed that when the inlet air temperature was 140 C., it was equipped with interlayer cold air. Astragalus polysaccharides slightly sticked to the wall, and spray drying was effective. The obtained Astragalus polysaccharides had a light color, as shown in
Experimental Example 2: 30-Day Feeding Experiment on Mice Using a Feeding Instant Astragalus Polysaccharide Immune-Enhancing Powder
[0035] Experiment method: 80 clean-grade ICR mice weighing 18-24 g were randomly divided into 4 groups, with 20 mice in each group, half male and half female. Experimental groups were divided into three dose groups: high, medium, and low, with doses of 1.0, 2.0, and 4.0 mg kg.sup.1 d.sup.1, respectively. Another control group was set up, and an equal dose of distilled water was given by gavage every day. General observation: during the experiment, the general state, physical signs, food intake, water consumption, and fecal status of mice were observed every day. The body weight was measured every 7 days, the dosage was adjusted according to changes in the body weight, and mice were continuously observed for 30 days. Measurement of hematology indicators: after 30 days of drug administration and 12 hours of fasting and water deprivation, eyeballs were removed and blood was taken to examine hematology indicators. Hematology examination items included: white blood cell count (WBC), percentage of neutrophils (NE %), percentage of lymphocytes (LY %), percentage of monocytes (MO %), percentage of eosinophils (EO %), basophils (BA %), red blood cell count (RBC), hemoglobin concentration (Hb), hematocrit (HCT %), mean corpuscular volume (MCV), mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width (RDW %), platelet count (PLT), and mean platelet volume (MPV). Measurement of blood biochemical indicators: After 30 days of drug administration and 12 hours of fasting and water deprivation, eyeballs were removed and blood was taken to examine blood biochemical indicators. The blood biochemical indicators included: glutamic pyruvic transaminase (ALT/GPT), glutamic oxalacetic transaminase (AST/GOT), total protein (TP), albumin (ALB), blood urea nitrogen (BUN), creatinine (CRE), total cholesterol (CHO), triglyceride (TG), and glucose (GLU). Observation of gross morphological changes: immediately after blood collection, organs such as the liver, kidney, spleen, stomach, testicles, or ovaries of the animals were dissected and weighed, and organ weight coefficients were calculated based on the body weight. Moreover, the morphological changes were observed with the naked eye.
[0036] Experiment Results:
[0037] General toxic reactions in mice: during the 30-day experimental period, the mice in the high, medium, and low dose administration groups and the control group showed normal activity, active behavior, glossy fur, normal food and water consumption, no abnormal changes in defecation and urination, and no deaths.
[0038] Body weight changes of mice: the changes in body weight after 30 days of feeding are shown in
[0039] Organ coefficients of mice: after 30 days of feeding, the organ coefficients of mice are shown in
[0040] Blood routine indicators: The results of the effects of the Astragalus polysaccharide powder on blood routine indicators in mice are shown in Table 3.
TABLE-US-00003 TABLE 3 Effects of the astragalus polysaccharide powder on blood routine in mice (
[0041] Table 3 showed no significant difference in white blood cell count (WBC), percentage of neutrophils (NE %), percentage of lymphocytes (LY %), percentage of monocytes (MO %), percentage of eosinophils (EO %), basophils (BA %), red blood cell count (RBC), hemoglobin concentration (Hb), hematocrit (HCT %), mean corpuscular volume (MCV), mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width (RDW %), platelet count (PLT), mean platelet volume (MPV) and other indicators between each dose group and the control group (P>0.05).
[0042] Blood biochemical indicators: Table 4 showed no significant difference in blood biochemical indicators such as glutamic pyruvic transaminase (GPT), glutamic oxalacetic transaminase (GOT), total protein (TP), albumin (ALB), blood urea nitrogen (BUN), creatinine (CRE), total cholesterol (CHO), triglyceride (TG), and glucose (GLU) between the mice in each dose group and the control group (P>0.05).
TABLE-US-00004 TABLE 4 Effects of the astragalus polysaccharide powder on blood biochemistry in Mice (
[0043] Pathological and histological examination of mice fed for 30 days: the size, shape, color, texture, etc. of organs such as the heart, liver, kidney, spleen, stomach, testicles, and ovaries in each group were comprehensively and carefully observed, and no abnormalities were found. There was no obvious degeneration or necrosis of the main organs, and their luster was normal.
[0044] The 30-day feeding experiment on mice showed that the feeding instant Astragalus polysaccharide immune-enhancing powder had high food safety.
Experimental Example 3: Immune Protection Experiment of the Feeding Instant Astragalus Polysaccharide Immune-Enhancing Powder
[0045] Experimental method: after one week of adaptation, the animals were randomly divided into groups based on gender and body weight. Thirty-six young mice were randomly divided into three groups: negative control group, model control group, and Astragalus polysaccharide administration group, with 12 mice in each group, half male and half female. A suspension of the Astragalus polysaccharide powder was prepared using water as a solvent and administered to mice by gavage once a day for 30 consecutive days. The dosage was 200 mg kg.sup.1 d.sup.1, the animals were weighed once a week, and the dosage was adjusted according to changes in body weight. The mice in the model group were intraperitoneally injected with cyclophosphamide at a dose of 80 mg/kg/d for three consecutive days. The administration experiment was carried out. On the 9th, 10th, 11th, 19th, 20th and 21st days after the experiment, the mice were strengthened once again, that is, cyclophosphamide was intraperitoneally injected at a dose of 80 mg kg.sup.1 d.sup.1.
[0046] Experiment Results
[0047] Effect of the Astragalus polysaccharide powder on the body weight of mice: the measurement results of the body weight of mice are shown in
[0048] Effect of the Astragalus polysaccharide powder on the organ coefficients of immune organs in mice: the results of the organ coefficient measurement of immune organs in mice are shown in
[0049] Effect of the Astragalus polysaccharide powder on -interferon in peripheral blood of mice: the ELISA method was used, and the operation method was carried out according to the instructions of the reagent kit. The detection results are shown in
[0050] Effect of the Astragalus polysaccharide powder on IgG in peripheral blood of mice: determination of serum IgG in peripheral blood of mice: the ELISA method was used, and the operation method was carried out according to the instructions of the reagent kit. The detection results are shown in
[0051] Immunoglobulin G (IgG) is synthesized in the spleen and lymph nodes. It has the highest content in serum (accounting for 75% of the amount of Ig), and has a long half-life in serum. It is mainly distributed in serum and interstitial fluid, and is the main component of anti-bacteria, anti-toxin and anti-virus antibodies, and also an important material basis in the process of anti-infection immunity of the body. Due to its highest content, IgG is the most important material basis for immune response. It has important immune effects and is the main force in the body's fight against infection.
[0052] Effect of the Astragalus polysaccharide powder on T lymphocyte subpopulations in peripheral blood of mice: the lymphocyte subpopulations were analyzed using a flow cytometer, and the proportion of cells in the cross doors was statistically analyzed using BD CellQuest Pro data analysis software. The results of the effect of the Astragalus polysaccharide powder on T lymphocyte subpopulations in peripheral blood of mice are shown in