CN111084231A - High-activity corn stigma compound polysaccharide and preparation method thereof - Google Patents
High-activity corn stigma compound polysaccharide and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a high-activity corn stigma compound polysaccharide and a preparation method thereof, belonging to the technical field of extraction and separation of natural plant active polysaccharides. The corn stigma polysaccharide, the hazel mushroom polysaccharide and the oyster mushroom polysaccharide are prepared by an ultrafiltration method, and then the composite polysaccharide is prepared according to a proportion, wherein the corn stigma ultrafiltration polysaccharide powder is hazel mushroom ultrafiltration polysaccharide powder, and the oyster mushroom ultrafiltration polysaccharide powder =1:0.2: 0.1-1: 0.8:0.6 and is mixed uniformly to obtain the corn stigma composite polysaccharide. The compound polysaccharide has the effect of promoting the proliferation of probiotics, namely lactic acid bacteria, in the yogurt better than three kinds of single polysaccharides. Meanwhile, compared with the traditional method for preparing the polysaccharide by water extraction and alcohol precipitation, the hazel mushroom polysaccharide and the oyster mushroom polysaccharide prepared by the method have higher extraction yield and better effect of promoting proliferation of probiotics-lactobacillus in the yoghourt.
Description
Technical Field
The invention relates to a high-activity corn stigma compound polysaccharide and a preparation method thereof, belonging to the technical field of extraction and separation of natural plant active polysaccharides.
Background
Corn silk (Stigmata madis) Corn (Yumai, Zimao, Zea mays, Poaceae)Zea mays L．) The style of the flower pillar is a traditional Chinese herbal medicine, is recorded in Yunnan Bencao and Chinese medicine dictionary, and is a common medicinal material variety recorded in 1985 edition (one part) of the ministry of health of the people's republic of China. China is a big country for corn production, and the annual yield of corn is 2 nd in the world. Thus, the corn silk, which is a byproduct of corn, is very abundant in resources.
The Armillaria mellea is fruiting body of Armillaria mellea of Eumycota. The mellea armillaria sporophore is smooth, tender and tasty, delicious in taste and rich in nutrition, grows on the dry base, the root generation, the fallen wood and the branches buried in the soil of the broad-leaved trees within 7-8 months, and is called as 'mountain delicacies' and 'the fourth treasure in northeast China'. The mellea armillaria sporophore is delicious, but the fresh mellea armillaria sporophore is short in preservation period, rich in nutrition, contains various amino acids and vitamins necessary for human bodies, and can strengthen the immunity of the human bodies, benefit intelligence, relieve mental anxiety, tonify qi, avoid hunger, prolong life and lighten the body and the like after being eaten frequently.
The oyster mushroom is famous and pleurotus ostreatus, is a wide variety of edible fungi cultivated in China, has the characteristics of low price and rich nutrition, and is well known to be consumed by the public.
The polysaccharide compound is an important active component, a large number of researches show that the polysaccharide has various effects of reducing blood sugar, regulating immunity, resisting bacteria, resisting tumors and the like, and clinical tests prove that the polysaccharide can also be used as an effective adjuvant therapy medicament for tumor chemotherapy and radiotherapy, and has good application prospect and development value in the aspects of medicines and foods.
The traditional polysaccharide extraction adopts an organic solvent precipitation method to remove small molecular impurities and prepare polysaccharide, such as 80% ethanol precipitation and the like. The method not only needs to consume a large amount of organic solvent, but also has the characteristics of strong volatilization, flammability, explosiveness and the like, so the method has higher requirements on equipment investment.
Disclosure of Invention
The invention provides a high-activity corn stigma compound polysaccharide and a preparation method thereof.
The technical scheme adopted by the invention for solving the technical problems is as follows: a high-activity corn stigma compound polysaccharide and a preparation method thereof comprise the following steps:
preparation of corn stigma polysaccharide
1. Pretreatment of corn silk
Removing impurities from stigma Maydis, oven drying at 60-70 deg.C, naturally cooling in dry environment, and pulverizing to 40-100 mesh;
2. defatting of corn stigma
Adding 95% ethanol or absolute ethanol into the corn stigma powder according to the mass-volume ratio of 1:3-1:5, soaking for 12-24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1-2 times by the same method, naturally drying in the shade, and storing in a dry environment for later use;
3. ultrafiltration polysaccharide preparation
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:15-1:30, and extracting for 0.5-3 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 5000 r/min, and separating the supernatant and the precipitate (corn stigma particles);
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10 KDa under ultrafiltration pressure of 0.05-2.0 MPa; an ultrafiltration method comprises supplementing 1/10-1 times of distilled water to the concentrated solution when the supernatant is ultrafiltered and concentrated to 1/20-1/2 times of the volume of the leaching solution, continuously ultrafiltering until the volume of the concentrated solution is recovered to the volume before supplementing water, repeating water supplementing in the same method until the total water volume is 2-5 times of the volume of the leaching solution, and no water is supplemented;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain the corn stigma ultrafiltration polysaccharide powder.
Extraction rate of the corn stigma ultrafiltration polysaccharide powder (%) = mass of the corn stigma ultrafiltration polysaccharide powder/mass of the corn stigma raw material x 100.
Second, preparation of mellea armillaria sporophore polysaccharide
1. Pretreatment of mellea armillaria sporophore
Pulverizing dried Armillaria mellea to 40-100 mesh;
2. degreasing hazel mushroom
Adding petroleum ether into the hazel mushroom powder according to the mass-to-volume ratio of 1:5-1:10, soaking for 12-24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1-2 times by the same method, naturally drying in the shade, and storing in a dry environment for later use;
3. preparation of mellea armillaria sporophore polysaccharide
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:20-1:40, and extracting for 0.5-3 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 8000 r/min, separating the supernatant and precipitate;
3.3 Re-extraction of the precipitate 1 time with the same method. Combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10-500 KDa under ultrafiltration pressure of 0.05-2.0 MPa; the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/20-1/2 times of the volume of the leaching liquor, supplementing distilled water into the concentrated solution, and keeping the volume of the concentrated solution constant until the volume of the total added water is 4-6 times of the volume of the leaching liquor;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain the hazel mushroom ultrafiltration polysaccharide powder.
The extraction rate of the hazel mushroom ultrafiltration polysaccharide powder is 5.36-7.02%, wherein the polysaccharide content is 32.01-42.28%.
The calculation formula is as follows:
the extraction rate of the hazel mushroom ultrafiltration polysaccharide powder (%) = the quality of the hazel mushroom ultrafiltration polysaccharide powder/the quality of the raw material of the hazel mushroom × 100.
Preparation of oyster mushroom polysaccharide
1. Pretreatment of oyster mushroom
Drying Pleurotus Ostreatus, and pulverizing to 40-100 mesh;
2. degreasing of oyster mushroom
Adding the oyster mushroom powder into anhydrous ether according to the mass volume ratio of 1:5-1:10, soaking for 12-24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1-2 times by the same method, naturally drying in the shade, and storing in a dry environment for later use;
3. preparation of oyster mushroom polysaccharide
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:20-1:40, and extracting for 0.5-3 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 8000 r/min, separating the supernatant and precipitate;
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10-500 KDa under ultrafiltration pressure of 0.05-2.0 MPa; the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/20-1/2 times of the volume of the leaching liquor, supplementing distilled water into the concentrated solution, and keeping the volume of the concentrated solution constant until the volume of the total added water is 4-6 times of the volume of the leaching liquor;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain Pleurotus ostreatus ultrafiltration polysaccharide powder.
The extraction yield of Pleurotus Ostreatus ultrafiltration polysaccharide powder is 4.97-7.16%, wherein the polysaccharide content is 33.10-44.65%.
The calculation formula is as follows:
extraction rate of oyster mushroom ultrafiltration polysaccharide powder (%) = mass of oyster mushroom ultrafiltration polysaccharide powder/mass of oyster mushroom raw material × 100.
Preparation of corn stigma compound polysaccharide
And (2) preparing the composite polysaccharide according to a ratio, and uniformly mixing the corn stigma ultrafiltration polysaccharide powder, the hazel mushroom ultrafiltration polysaccharide powder, the oyster mushroom ultrafiltration polysaccharide powder =1:0.2: 0.1-1: 0.8:0.6 to obtain the corn stigma composite polysaccharide.
The invention has the beneficial effects that: the high-activity corn stigma compound polysaccharide prepared by the invention is prepared by mixing three components of polysaccharide according to a proportion. The compound polysaccharide has the effect of promoting the proliferation of probiotics, namely lactic acid bacteria, in the yogurt better than three kinds of single polysaccharides. Meanwhile, compared with the traditional method for preparing the polysaccharide by water extraction and alcohol precipitation, the hazel mushroom polysaccharide and the oyster mushroom polysaccharide prepared by the method have higher extraction yield and better effect of promoting proliferation of probiotics-lactobacillus in the yoghourt.
Drawings
FIG. 1 is a graph of data showing the effect of adding different types of stigma Maydis polysaccharides on the total number of lactobacillus in yogurt.
FIG. 2 is a graph of data showing the effect of different mellea armillaria sporophore polysaccharides on the total number of lactic acid bacteria in yogurt.
FIG. 3 is a data chart showing the effect of different types of Pleurotus ostreatus polysaccharide on the total number of lactic acid bacteria in yogurt.
FIG. 4 is a data chart of the effect of corn stigma ultrafiltration polysaccharide, mellea armillaria ultrafiltration polysaccharide, oyster mushroom ultrafiltration polysaccharide and corn stigma composite polysaccharide prepared by compounding three polysaccharides on the total number of lactic acid bacteria in yogurt;
1-blank control (no addition); 2-corn stigma ultrafiltration polysaccharide; 3-mellea armillaria sporophore ultrafiltration polysaccharide; 4-oyster mushroom ultrafiltration polysaccharide; 5-complex polysaccharide 1 (1: 0.2: 0.1); 6-complex polysaccharide 2 (1: 0.5: 0.35); 7-complex polysaccharide 3 (1: 0.2: 0.6); 8-complex polysaccharide 4 (1: 0.8: 0.1); 9-complex polysaccharide 5 (1: 0.8: 0.6); 10-complex polysaccharide 6 (1: 0.1: 0.1); 11-complex polysaccharide 7 (1: 0.2: 0.05); 12-complex polysaccharide 8 (1: 0.8: 0.1); 13-complex polysaccharide 9 (1: 0.2: 0.7); 14-Complex polysaccharide 10 (1: 1: 1).
Detailed Description
The invention is further described with reference to the following figures and detailed description.
Example 1
A high-activity corn stigma compound polysaccharide and a preparation method thereof comprise the following steps:
preparation of corn stigma polysaccharide
1. Pretreatment of corn silk
Removing impurities from stigma Maydis, oven drying at 60 deg.C, naturally cooling in dry environment, and pulverizing to 80 mesh;
2. defatting of corn stigma
1 kg of corn stigma powder, adding absolute ethyl alcohol according to the mass-to-volume ratio of 1:5, soaking for 12 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1 time by the same method, naturally drying in the shade, and storing in a dry environment for later use;
3. ultrafiltration polysaccharide preparation
3.1 adding 20L of distilled water into the extraction according to the mass volume ratio of 1:20, and extracting for 2 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 5000 r/min, and separating the supernatant and the precipitate (corn stigma particles);
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10 KDa under ultrafiltration pressure of 0.08 MPa;
the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/10 times (2L) of the leaching liquor, 1L of distilled water with 1/2 times of the volume of the concentrated solution is supplemented into the concentrated solution, the ultrafiltration is continued until the volume of the concentrated solution is recovered to the volume before water supplementation, and water supplementation is repeated in the same way until the total water addition volume is 4 times (80L) of the volume of the leaching liquor, and water is not supplemented any more;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain the corn stigma ultrafiltration polysaccharide powder.
Second, preparation of mellea armillaria sporophore polysaccharide
1. Pretreatment of mellea armillaria sporophore
Pulverizing dried Armillaria mellea to 60 mesh;
2. degreasing hazel mushroom
Adding the mellea armillaria sporophore powder into petroleum ether according to the mass-to-volume ratio of 1:5, soaking for 24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1 time by the same method, naturally drying in a shade, and storing in a dry environment for later use;
3. preparation of mellea armillaria sporophore polysaccharide:
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:20, and extracting for 1.5 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 8000 r/min, separating the supernatant and precipitate;
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10 KDa and ultrafiltration pressure of 0.10 Mpa;
the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/8 times of the volume of the leaching liquor, supplementing distilled water into the concentrated solution, and keeping the volume of the concentrated solution constant until the volume of the total added water is 4 times of the volume of the leaching liquor;
3.6, drying: continuously ultrafiltering the concentrated solution until dry matter content reaches 15-20%, and freeze drying or spray drying to obtain Hazel mushroom ultrafiltration polysaccharide powder.
Preparation of oyster mushroom polysaccharide
1. Pretreatment of oyster mushroom
Drying oyster mushroom and crushing to 60 meshes;
2. degreasing of oyster mushroom
Adding the oyster mushroom powder into anhydrous ether according to the mass volume ratio of 1:5, soaking for 24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1 time by the same method, naturally drying in a shade, and storing in a dry environment for later use;
3. preparation of oyster mushroom polysaccharide
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:20, and extracting for 2 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 8000 r/min, separating the supernatant and precipitate;
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10 KDa under ultrafiltration pressure of 0.08 MPa;
the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/8 times of the volume of the leaching liquor, supplementing distilled water into the concentrated solution, and keeping the volume of the concentrated solution constant until the volume of the total added water is 4 times of the volume of the leaching liquor;
3.6, drying: continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain Pleurotus Ostreatus ultrafiltration polysaccharide powder.
Preparation of corn stigma compound polysaccharide
Preparing corn stigma composite polysaccharide according to a proportion, and uniformly mixing corn stigma ultrafiltration polysaccharide powder, hazel mushroom ultrafiltration polysaccharide powder, oyster mushroom ultrafiltration polysaccharide powder =1:0.2:0.1 to obtain corn stigma composite polysaccharide 1;
preparing corn stigma composite polysaccharide according to a proportion, and uniformly mixing corn stigma ultrafiltration polysaccharide powder, hazel mushroom ultrafiltration polysaccharide powder, oyster mushroom ultrafiltration polysaccharide powder =1:0.5:0.35 to obtain corn stigma composite polysaccharide 2;
preparing corn stigma composite polysaccharide according to a proportion, and uniformly mixing corn stigma ultrafiltration polysaccharide powder, hazel mushroom ultrafiltration polysaccharide powder, oyster mushroom ultrafiltration polysaccharide powder =1:0.2:0.6 to obtain corn stigma composite polysaccharide 3;
preparing corn stigma composite polysaccharide according to a proportion, and uniformly mixing corn stigma ultrafiltration polysaccharide powder, hazel mushroom ultrafiltration polysaccharide powder, oyster mushroom ultrafiltration polysaccharide powder =1:0.8:0.1 to obtain corn stigma composite polysaccharide 4;
preparing the corn stigma composite polysaccharide according to the proportion, and uniformly mixing the corn stigma ultrafiltration polysaccharide powder, the hazel mushroom ultrafiltration polysaccharide powder, the oyster mushroom ultrafiltration polysaccharide powder =1:0.8:0.6 to obtain the corn stigma composite polysaccharide 5.
The extraction rate of the corn stigma ultrafiltration polysaccharide powder is 7.16%, wherein the polysaccharide content is 35.98%; the extraction rate of the mellea armillaria sporophore polysaccharide powder is 6.86 percent, wherein the polysaccharide content is 38.21 percent, and the extraction rate of the polysaccharide is improved by 10 to 35 percent compared with the extraction rate of the polysaccharide by the traditional water extraction and alcohol precipitation method; the extraction rate of the oyster mushroom polysaccharide powder is 6.98 percent, wherein the polysaccharide content is 37.55 percent, and the extraction rate of the polysaccharide is improved by 8 to 30 percent compared with the traditional water extraction and alcohol precipitation method.
Example 2
Polysaccharide for promoting proliferation of probiotics in yogurt
1. Adding raw milk into sterile beaker (100 mL/cup) in aseptic room for preparing yogurt, adding 1% experimental polysaccharide, adding blank control group (not adding), repeating for 3 times in each experiment, and adding mixed probioticThe strains (mixed strains of Streptococcus thermophilus, Lactobacillus bulgaricus, Bifidobacterium lactis, and Lactobacillus acidophilus with total viable count of 1 × 106CFU/mL), the inoculum size is 16.7%, the probiotic bacteria are cultured for 8 h at 45 ℃ after sealing, and the proliferation condition of the probiotic bacteria is detected.
2. Lactic acid bacteria detection
(1) Sample dilution
Under aseptic condition, taking 1mL of sample to be detected in aseptic operation, adding the sample to a sterile test tube filled with 9mL of sterile physiological saline, and shaking uniformly to prepare a sample diluent with the ratio of 1:10 (mL/mL). And taking 1mL of the sample diluent with the volume ratio of 1:10 (mL/mL) by using a micropipette, injecting the sample diluent into a sterile test tube filled with 9mL of sterile physiological saline, repeatedly blowing and beating the sample diluent to be uniformly mixed to prepare the sample diluent with the volume ratio of 1:100 (mL/mL), and sequentially preparing the sample diluent with the volume ratio of 10 times increased.
(2) Lactic acid bacteria count
The method is carried out according to the GB 4789.35-2010 national standard for food safety-food microbiology inspection-lactobacillus inspection method. Briefly described, the following steps: according to the estimation of the total viable count of the sample to be detected, 2-3 continuous suitable dilutions are selected, 0.1mL of sample is taken from each dilution, the samples are respectively placed on 3 MRS agar plates, and the surfaces of the samples are coated by using a coating rod. Counting all colonies on the plate after anaerobic culture for 48 h +/-2 h at 36 +/-1 ℃.
As can be seen from the figure 1, the corn stigma ultrafiltration polysaccharide prepared by the method can obviously promote the proliferation of probiotics-lactic acid bacteria in the yoghourt, and the corn stigma alcohol precipitation polysaccharide prepared by the traditional water extraction and alcohol precipitation method also has the function of promoting the proliferation of the probiotics-lactic acid bacteria in the yoghourt. Compared with the alcohol precipitation polysaccharide prepared by the traditional method, the corn stigma ultrafiltration polysaccharide prepared by the invention has better proliferation promoting effect on probiotics-lactic acid bacteria in the yoghourt.
As can be seen from the figure 2, the addition of the mellea armillaria sporophore ultrafiltration polysaccharide prepared by the method can obviously promote the proliferation of probiotics-lactic acid bacteria in the yoghourt, and the addition of the mellea armillaria sporophore alcohol precipitation polysaccharide prepared by the traditional water extraction and alcohol precipitation method also has the effect of promoting the proliferation of the probiotics-lactic acid bacteria in the yoghourt. Compared with the alcohol precipitation polysaccharide prepared by the traditional method, the hazel mushroom ultrafiltration polysaccharide prepared by the invention has better proliferation promoting effect on probiotics-lactic acid bacteria in the yoghourt.
As can be seen from the figure 3, the oyster mushroom ultrafiltration polysaccharide prepared by the method can obviously promote the proliferation of probiotics-lactic acid bacteria in the yogurt, and the oyster mushroom alcohol precipitation polysaccharide prepared by the traditional water extraction and alcohol precipitation method also has the effect of promoting the proliferation of the probiotics-lactic acid bacteria in the yogurt. Compared with the alcohol precipitation polysaccharide prepared by the traditional method, the oyster mushroom ultrafiltration polysaccharide prepared by the invention has better proliferation promoting effect on probiotics-lactic acid bacteria in yoghourt.
Referring to fig. 4, the corn stigma ultrafiltration polysaccharide, the hazel mushroom ultrafiltration polysaccharide, the oyster mushroom ultrafiltration polysaccharide and the corn stigma composite polysaccharide prepared by compounding the three polysaccharides are obviously superior to blank control (no additive) in the proliferation promotion effect of probiotics-lactobacillus in the yogurt. Compared with 3 original polysaccharides, the composite polysaccharide 1-5 (namely corn stigma ultrafiltration polysaccharide powder: hazel mushroom ultrafiltration polysaccharide powder: oyster mushroom ultrafiltration polysaccharide powder =1:0.2: 0.1-1: 0.8: 0.6) has better proliferation promoting effect, but when the compounding ratio of the 3 polysaccharides is not in the range, the proliferation promoting effect is not obviously improved, and the activity of the composite polysaccharide 6-10 is not different from that of the 3 original polysaccharides, and is also obviously lower than that of the composite polysaccharide 1-5. Therefore, the 3 polysaccharides have synergistic effect, can enhance the proliferation promoting effect on the probiotics-lactobacillus in the yoghourt, but the synergistic effect exists only in a certain proportion range.
Claims (3)
1. A high-activity corn stigma compound polysaccharide and a preparation method thereof are characterized in that: the method comprises the following steps:
preparation of corn stigma polysaccharide
1. Pretreatment of corn silk
Removing impurities from stigma Maydis, oven drying at 60-70 deg.C, naturally cooling in dry environment, and pulverizing to 40-100 mesh;
2. defatting of corn stigma
Adding 95% ethanol or absolute ethanol into the corn stigma powder according to the mass-volume ratio of 1:3-1:5, soaking for 12-24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1-2 times by the same method, naturally drying in the shade, and storing in a dry environment for later use;
3. ultrafiltration polysaccharide preparation
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:15-1:30, and extracting for 0.5-3 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 5000 r/min, and separating the supernatant and the precipitate;
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10 KDa under ultrafiltration pressure of 0.05-2.0 MPa; an ultrafiltration method comprises supplementing 1/10-1 times of distilled water to the concentrated solution when the supernatant is ultrafiltered and concentrated to 1/20-1/2 times of the volume of the leaching solution, continuously ultrafiltering until the volume of the concentrated solution is recovered to the volume before supplementing water, repeating water supplementing in the same method until the total water volume is 2-5 times of the volume of the leaching solution, and no water is supplemented;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain stigma Maydis ultrafiltration polysaccharide powder;
second, preparation of mellea armillaria sporophore polysaccharide
1. Pretreatment of mellea armillaria sporophore
Pulverizing dried Armillaria mellea to 40-100 mesh;
2. degreasing hazel mushroom
Adding petroleum ether into the hazel mushroom powder according to the mass-to-volume ratio of 1:5-1:10, soaking for 12-24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1-2 times by the same method, naturally drying in the shade, and storing in a dry environment for later use;
3. preparation of mellea armillaria sporophore polysaccharide
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:20-1:40, and extracting for 0.5-3 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 8000 r/min, separating the supernatant and precipitate;
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10-500 KDa under ultrafiltration pressure of 0.05-2.0 MPa; the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/20-1/2 times of the volume of the leaching liquor, supplementing distilled water into the concentrated solution, and keeping the volume of the concentrated solution constant until the volume of the total added water is 4-6 times of the volume of the leaching liquor;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain the hazel mushroom ultrafiltration polysaccharide powder;
preparation of oyster mushroom polysaccharide
1. Pretreatment of oyster mushroom
Drying Pleurotus Ostreatus, and pulverizing to 40-100 mesh;
2. degreasing of oyster mushroom
Adding the oyster mushroom powder into anhydrous ether according to the mass volume ratio of 1:5-1:10, soaking for 12-24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1-2 times by the same method, naturally drying in the shade, and storing in a dry environment for later use;
3. preparation of oyster mushroom polysaccharide
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:20-1:40, and extracting for 0.5-3 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 8000 r/min, separating the supernatant and precipitate;
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10-500 KDa under ultrafiltration pressure of 0.05-2.0 MPa; the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/20-1/2 times of the volume of the leaching liquor, supplementing distilled water into the concentrated solution, and keeping the volume of the concentrated solution constant until the volume of the total added water is 4-6 times of the volume of the leaching liquor;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain Pleurotus ostreatus ultrafiltration polysaccharide powder;
preparation of corn stigma compound polysaccharide
And (2) preparing the composite polysaccharide according to a ratio, and uniformly mixing the corn stigma ultrafiltration polysaccharide powder, the hazel mushroom ultrafiltration polysaccharide powder, the oyster mushroom ultrafiltration polysaccharide powder =1:0.2: 0.1-1: 0.8:0.6 to obtain the corn stigma composite polysaccharide.
2. The high-activity corn silk complex polysaccharide and the preparation method thereof as claimed in claim 1, wherein the high-activity corn silk complex polysaccharide is characterized in that: the preparation method of the mellea armillaria sporophore polysaccharide comprises the following steps:
(1) pretreatment of mellea armillaria sporophore
Pulverizing dried Armillaria mellea to 40-100 mesh;
(2) degreasing hazel mushroom
Adding petroleum ether into the hazel mushroom powder according to the mass-to-volume ratio of 1:5-1:10, soaking for 12-24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1-2 times by the same method, naturally drying in the shade, and storing in a dry environment for later use;
(3) preparation of mellea armillaria sporophore polysaccharide
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:20-1:40, and extracting for 0.5-3 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 8000 r/min, separating the supernatant and precipitate;
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10-500 KDa under ultrafiltration pressure of 0.05-2.0 MPa; the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/20-1/2 times of the volume of the leaching liquor, supplementing distilled water into the concentrated solution, and keeping the volume of the concentrated solution constant until the volume of the total added water is 4-6 times of the volume of the leaching liquor;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain the hazel mushroom ultrafiltration polysaccharide powder;
the extraction yield is 5.36-7.02%, wherein the polysaccharide content is 32.01-42.28%.
3. The high-activity corn silk complex polysaccharide and the preparation method thereof as claimed in claim 1, wherein the high-activity corn silk complex polysaccharide is characterized in that: the preparation method of the oyster mushroom polysaccharide is characterized by comprising the following steps: the method comprises the following steps:
(1) pretreatment of oyster mushroom
Drying Pleurotus Ostreatus, and pulverizing to 40-100 mesh;
(2) degreasing of oyster mushroom
Adding the oyster mushroom powder into anhydrous ether according to the mass volume ratio of 1:5-1:10, soaking for 12-24 hours, filtering under reduced pressure, repeatedly soaking the filter residue for 1-2 times by the same method, naturally drying in the shade, and storing in a dry environment for later use;
(3) preparation of oyster mushroom polysaccharide
3.1 adding distilled water into the extraction according to the mass volume ratio of 1:20-1:40, and extracting for 0.5-3 hours in an extraction tank at the temperature of 95-100 ℃;
3.2 centrifuging the centrifugal extract at 8000 r/min, separating the supernatant and precipitate;
3.3 extracting the precipitate again for 1 time by the same method, and combining the supernatants obtained by the two extractions;
3.4 removing impurities and filtering the supernatant through paper to remove micro particles and prepare polysaccharide extraction clear solution;
3.5 ultrafiltering the supernatant with ultrafiltration membrane with molecular weight cutoff of 10-500 KDa under ultrafiltration pressure of 0.05-2.0 MPa; the ultrafiltration method comprises the following steps: when the supernatant is concentrated by ultrafiltration to 1/20-1/2 times of the volume of the leaching liquor, supplementing distilled water into the concentrated solution, and keeping the volume of the concentrated solution constant until the volume of the total added water is 4-6 times of the volume of the leaching liquor;
3.6 drying, continuously ultrafiltering the concentrated solution until the dry matter content reaches 15-20%, and freeze drying or spray drying to obtain Pleurotus ostreatus ultrafiltration polysaccharide powder;
the extraction yield is 4.97-7.16%, wherein the polysaccharide content is 33.10-44.65%.
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| CN115088846A (en) * | 2022-05-31 | 2022-09-23 | 十堰市武当山旅游经济特区净界健康管理有限公司 | Composite millet bacteria polysaccharide oligomer powder for enhancing immune function and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115088846A (en) * | 2022-05-31 | 2022-09-23 | 十堰市武当山旅游经济特区净界健康管理有限公司 | Composite millet bacteria polysaccharide oligomer powder for enhancing immune function and preparation method thereof |
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