CN108477612B - Dendrobium nutrition powder containing dendrobium officinale purified extract and preparation method thereof - Google Patents
Dendrobium nutrition powder containing dendrobium officinale purified extract and preparation method thereof Download PDFInfo
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- CN108477612B CN108477612B CN201810117982.0A CN201810117982A CN108477612B CN 108477612 B CN108477612 B CN 108477612B CN 201810117982 A CN201810117982 A CN 201810117982A CN 108477612 B CN108477612 B CN 108477612B
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- polysaccharide
- dendrobium
- dendrobium officinale
- ethanol
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Classifications
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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
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/175—Amino acids
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention provides dendrobium nutrition powder containing a purified extract of dendrobium officinale and a preparation method thereof, wherein the nutrition powder particles have a core-shell structure body and at least comprise: amino acid modified slow-digestion starch of dendrobium officinale, purified polysaccharide of dendrobium officinale and dendrophenol. The dendrobium nutrition powder is suitable for various crowds including hyperglycemia, and has good hypoglycemic effect and healthy nutritive value.
Description
Technical Field
The invention relates to dendrobium nutrition powder capable of reducing blood sugar and containing a purified dendrobium officinale extract and a preparation method thereof, in particular to soluble dendrobium nutrition powder with a core-shell structure and containing specific polysaccharide components of dendrobium officinale, amino acids and a dendrophenol extract and a preparation method thereof, and belongs to the fields of blood sugar reducing foods and nutritional health care products.
Background
Dendrobe (school name: Dendrobium nobile Lindl), also named as herba Dendrobii orchid Yun, Nelumbo nucifera, herba Hylotelephii Erythrosticti, Zine immortal plant, herba Lini, Nelumbo Nucifera, herba Dendrobii, etc. The medicinal plants, sweet, light and slightly salty in nature and taste, are cold and enter stomach, kidney and lung meridians. To nourish stomach, promote the production of body fluid, nourish yin and clear heat. It is commonly used for yin deficiency and fluid deficiency, dry mouth with polydipsia, poor appetite, retching, and dim and unclear vision after illness. As traditional famous and precious Chinese medicinal materials in China, the medicinal dendrobium comprises fresh dendrobium, dendrobium nobile, dendrobium officinale, dendrobium candidum, dendrobium fimbriatum and the like; the 2010 version of pharmacopoeia begins to list the item "Dendrobium officinale" individually, which is a fresh or dry stem of Dendrobium officinale belonging to the genus Dendrobium of the family Orchidaceae. The item of 'dendrobium nobile' in the 2010 version of pharmacopoeia is divided into fresh or dry stems of three plants, namely dendrobium nobile, dendrobium chrysotoxum and dendrobium fimbriatum. In addition, the commonly used medicinal dendrobium can also be dendrobium huoshanense which is commonly called dendrobium huoshanense; herba Dendrobii, and herba Dendrobii.
Research medicines show that the dendrobium has extremely high medicinal and health-care functions, so that the dendrobium is deeply and widely researched. The research shows that the effective components of the medicine comprise: dendrobe polysaccharide, dendrobe amino acid, alkaloid, flavonoid and the like. The main drug effect component of the dendrobium polysaccharide is dendrobium polysaccharide which is one of active polysaccharides, and the dendrobium polysaccharide is a heteropolysaccharide with wider molecular weight distribution. The heteropolysaccharide is a polysaccharide composed of two or more different monosaccharide molecules. The study proves that the dendrobium polysaccharide can obviously reduce the blood sugar value of the organism, has no reducing effect on normal blood sugar and does not improve the level of serum insulin and has repairing effect on islet cells unlike oral hypoglycemic drugs such as sulfonylurea; meanwhile, the dendrobium officinale polysaccharide also has the function of reducing blood sugar outside pancreas, and can reduce the source of blood sugar, increase the way of blood sugar and achieve the purpose of reducing the blood sugar value by inhibiting hepatic glycogen decomposition caused by epinephrine and promoting hepatic glycogen synthesis.
Wushushu and the like at Zhejiang university perform special experimental research on the blood sugar reducing effect and mechanism of dendrobium officinale. Normal mice, adrenal disorder hyperglycemic mice and streptozotocin diabetes rats are selected as experimental objects, starting from blood sugar values, serum insulin and insulin levels, and by adopting methods such as radioimmunoassay and immunohistochemistry HRP-SPA chromosome and the like, the dendrobium officinale is determined to have obvious blood sugar reducing effect by combining morphological changes of islet alpha cells and beta cells and the like. The internal mechanism of the pancreas for reducing blood sugar is to promote insulin secretion of islet beta cells and inhibit glucagon secretion of islet alpha cells, and the external mechanism is to promote liver glycogen synthesis. The research provides pharmacological basis for the dendrobium officinale to treat diabetes, and shows wide prospect for the development of the dendrobium officinale as a new hypoglycemic drug.
Therefore, in the medicinal dendrobium, the dendrobium officinale contains high-quality polysaccharide content.
Further multiple researches find that the polysaccharide is the main component of the dendrobium officinale, and the content of the water-soluble polysaccharide is up to more than 22%. The strength of the physiological activity of the dendrobium is closely related to the polysaccharide content of the dendrobium. The higher the polysaccharide content, the heavier the quality, the more viscous the chewing, and the better the quality. The mass fractions of the total polysaccharide at different parts of the dendrobium officinale are sequentially from high to low, namely the middle stem, the upper stem, the lower stem and the root. However, the distribution rule of the water-soluble polysaccharide is that the root of the leaf and the root of the stem are larger than the stem. Modern researches find that the water-soluble polysaccharide of the dendrobium candidum protocorm mainly comprises mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose, and the weight ratio of molecular substances is 1.1204: 1: 1.046: 23.354: 3.828: 1.046. wherein, after the water-soluble polysaccharide is extracted, glucose occupies a huge part.
Therefore, in the prior art, the dendrobium officinale milled powder or the polysaccharide extract is directly eaten for expecting the method for reducing blood sugar, so that a large amount of components such as glucose and the like which are easy to cause hyperglycemia are often ingested, the medicinal effect of the dendrobium polysaccharide is greatly reduced, and even no corresponding effect exists. The invention also aims to overcome one of the defects of the dendrobium polysaccharide health care product in the prior art.
In the prior art, although many progresses are made on the research of pharmacological activity and chemical structure of the medicinal dendrobium polysaccharide, in the aspect of pharmacological activity research, the used polysaccharide is mainly crude polysaccharide, and the research on the active action mechanism of the medicinal dendrobium polysaccharide is not much; in the aspect of chemical structure research, the chemical structure of many polysaccharide components with pharmacological activity is not clear. Moreover, the separation and purification of the medicinal or food-grade dendrobium polysaccharide still has various defects, such as low extraction rate, low purity, complicated procedures, unsuitability for industrial production, serious waste of raw materials and the like.
In addition, the dendrobium has good effect on diabetic complications and complications. For example, the dendrophenol component of dendrobe is helpful for improving diabetes and cataract.
According to the research on the dendrophenol mechanism, the dendrophenol can block the expression of induced nitric oxide synthase and cyclooxygenase induced by lipopolysaccharide, is an anti-cataract effective component in the dendrobe, and can effectively inhibit AR (aldose reductase), improve the abnormal polyol metabolic pathway of a diabetic patient and prevent and delay the occurrence and development of diabetes and complications thereof, namely cataract.
Chendan and the like (university of traditional Chinese medicine in Guangzhou, 2014) research shows that the inhibitory rate of the dendrophenol with the concentration of 120 mu mol/L on iNOS is 100%, and the median inhibitory amount (IC50) is (32.227 +/-5.73) mu mol/L; dendrophenol inhibits the generation of NO by directly acting on iNOS active sites, and acting amino acid sites of the dendrophenol and the iNOS active sites are lle195 and Gln 257.
Other beneficial main components in dendrobium nobile also comprise amino acid. The amino acids in dendrobium stem exist mainly in free form, and the content of the amino acids is far higher than that of other components except polysaccharide. Free amino acid is also one of the main effective components in dendrobium. Experiments prove that the dendrobium officinale contains all essential amino acids of human bodies except tryptophan, wherein the main amino acids are aspartic acid, glutamic acid, glycine, valine and leucine, and the 5 nitrogen-based acids account for about 53 percent of the total amino acids. Wherein, the content of glutamic acid, aspartic acid and glycine accounts for about 35.8 percent of the total content.
At present, more extraction processes and preparation products for dendrobium have been disclosed, such as:
CN104784505A discloses a dendrobium officinale health product assisting in reducing blood sugar and a preparation method thereof, wherein the dendrobium officinale health product comprises, by weight, 35-45 parts of fresh dendrobium officinale strips, 1-2 parts of American ginseng, 2-4 parts of kudzu vine roots, 2-4 parts of radices trichosanthis and 1-2 parts of schisandra chinensis. The preparation method comprises the following steps: weighing raw materials, processing fresh dendrobium officinale strips, processing American ginseng, extracting with alcohol, extracting with water, drying and mixing. The raw material formula traditional Chinese medicine has the effects of nourishing yin, benefiting qi, clearing heat and promoting fluid production, and the effective components of the medicine are easy to absorb and are beneficial to exerting the drug effect; all medicinal materials in the raw material formula are medicinal and edible medicinal materials, and have no toxic or side effect.
CN104126746A discloses a dendrobium officinale composition for reducing blood sugar and blood fat and a preparation method thereof, wherein the dendrobium officinale composition is prepared from the following traditional Chinese medicine raw materials in parts by weight: 10-40 parts of dendrobium officinale, 10-30 parts of Chinese yam, 5-35 parts of semen cassiae, 10-30 parts of rhizoma polygonati, 10-30 parts of oat extract, 10-20 parts of maltodextrin and 2-10 parts of soybean oligosaccharide. The invention has the advantages that: the invention comprehensively utilizes various raw materials such as dendrobium officinale, Chinese yam, cassia seed, rhizoma polygonati, oat beta-glucan, soybean oligosaccharide and the like, and is supplemented with a proper amount of maltodextrin to prepare the composition with the function of reducing blood sugar and blood fat. The dendrobium officinale, the Chinese yam, the cassia seed, the rhizoma polygonati and the oat all contain polysaccharide substances, and the practices prove that the dendrobium officinale, the Chinese yam, the cassia seed, the rhizoma polygonati and the oat have good effects of reducing blood sugar and blood fat, treating diabetes and the like.
CN104958564A discloses a dendrobium officinale oral liquid and a preparation method thereof, the dendrobium officinale oral liquid is prepared by decocting dendrobium officinale, arillus longan and mulberry with water, and every 1000mL of the dendrobium officinale oral liquid is prepared by decocting 90-100 g of dendrobium officinale, 90-100 g of arillus longan and 90-100 g of mulberry with water. According to the invention, the raw materials are subjected to a preparation method of repeated decoction, effective components in medicinal materials can be efficiently extracted, three components including dendrobium officinale, arillus longan and mulberry in the dendrobium officinale oral liquid are blended in proportion, the effects of strengthening body resistance and cultivating primordial qi, nourishing yin and tonifying are achieved, natural plant polysaccharide substances contained in the components can enhance the immune function, the dendrobium officinale oral liquid is free of sucrose and any additive, and the dendrobium officinale oral liquid can be eaten by people with hyperglycemia and hyperlipidemia.
CN104324244A discloses a compound dendrobium officinale hypoglycemic capsule, which is prepared by adopting 7 traditional Chinese medicines of dendrobium officinale, astragalus, kudzu root, dried rehmannia root, trichosanthes root, rhizoma polygonati, medlar and red paeony root as auxiliary materials and adopting the steps of crushing, extracting, mixing, granulating and packaging, and has the functions of mutual influence and synergistic interaction; can promote insulin secretion of islet beta cells, inhibit glucagon secretion of islet alpha cells, inhibit hepatic glycogen decomposition caused by epinephrine and promote hepatic glycogen synthesis, thereby reducing blood sugar source, increasing blood sugar route, reducing blood sugar value, having no toxic or side effect and adverse reaction, and effectively controlling and eliminating the 'more than three or less' symptoms of diabetes and diabetic complications.
However, the above prior art generally uses whole dendrobium powder, which contains many components that are not beneficial for reducing blood sugar, as described above. Even if a few polysaccharides are extracted, the polysaccharide components are usually extracted by the prior art, the residue and the waste residues are discarded, and the amino acid in the residue and the waste residues are also wasted, so that the method is not economical.
The average mass fraction of total amino acids in the leaves of 11 dendrobium officinale lines is 7.43%, the stem content is 1.91%, the leaves are obviously higher than the stem, the amino acid composition is close to the standard of ideal protein provided by FAO/WHO, and the dendrobium officinale leaf extract has a good functional food development prospect.
There are many methods for extracting free amino acids, including ultrasonic extraction, enzymatic extraction, high-pressure hot water extraction, solvent extraction, microwave extraction, and the like. At present, a high-pressure hot water extraction method, an ultrasonic extraction method, an enzymolysis extraction method and the like are more applied, but the extraction rate and the extraction process of free amino acid still need to be improved, particularly, the extraction process of dendrobe amino acid reports are less, and the aim of comprehensively and fully utilizing the effective components of dendrobe is not achieved.
In addition, it is noted that the dendrobine in the whole dendrobe powder has the effects of relieving pain and fever, reducing heart rate and blood pressure, and slowing respiration, but can generate moderate hyperglycemia. In addition, dendrobine is an alkaloid and is not suitable for overdose. Dendrobine is distributed in, for example, Dendrobium nobile, Dendrobium officinale, Dendrobium primrose, Dendrobium loddigesii, Dendrobium bracteatum, Dendrobium tenuiflorum, and Dendrobium nobile. At present, dendrobium powder in the market mostly contains dendrobine, which is particularly useless for patients with hyperglycemia, and particularly, the blood sugar rise is easy to aggravate when the dendrobium powder is taken excessively or frequently. Therefore, for patients with hyperglycemia, the separation and extraction of beneficial components in dendrobium stem and the addition of a proper preparation form are urgently needed.
Another problem existing in the dendrobium health care product composition for reducing blood sugar in the prior art is that no proper matched carrier exists. On one hand, when other hypoglycemic traditional Chinese medicine component extracts are combined, the mouth feel is poor, such as bitter and astringent, the dosage is often required to be controlled, and hypoglycemia and syncope even caused by excessive administration are avoided; on the other hand, when non-medicinal components such as yam, grains and the like are combined, excessive starch or carbohydrate ingestion is often caused, the catabolism of the substances in the body indirectly raises the blood sugar level, and the effect is greatly reduced, so that an urgent need also exists for a proper carrier or component matched with dendrobe polysaccharide.
Slowly Digestible Starch (SDS), which refers to starch that is absorbed in the small intestine by complete digestion but at a slower rate (20-120 min); in recent years, clinical researches show that the slowly-digested starch has special physiological functions of slowly digesting and absorbing, continuously releasing energy, maintaining postprandial blood sugar steady state, improving glucose tolerance, reducing postprandial insulin secretion, improving the sensitivity of the organism to insulin and preventing and treating various chronic diseases related to diet. The slowly digestible starch is used as a novel functional product, has two requirements of nutrition and cure function of diabetes, not only conforms to the trend of scientific and technological development of modern food and medicine, but also meets the health requirements of consumers, and has important significance for reasonably utilizing carbohydrate to prevent metabolic syndrome related to insulin resistance. Therefore, many researchers have focused on the study of slowly digestible starch. For example, patent application No. 201110112582.9 discloses placing prolamin in a beaker, adding ethanol, stirring in a water bath until prolamin is completely dissolved, adding oleic acid, stirring well, adding corn starch, and continuing stirring. Homogenizing for 3 times in a high pressure homogenizer, centrifuging, spray drying to obtain slowly digestible starch product. The patent application No. 201310058866.3 uses pullulanase and glucosyltransferase to treat wheat starch and prepare slowly digestible starch by repeated recrystallization.
As described above, although the prior art provides various methods for preparing slowly digestible starch, the content of the slowly digestible starch prepared is still not high enough and the efficacy and composition are single.
Disclosure of Invention
As mentioned above, in order to overcome the defects of the dendrobium officinale food composition for reducing blood sugar in the prior art, the invention aims to provide dendrobium officinale nutritional powder which has a core-shell structure and good taste and can reduce blood sugar and a preparation method thereof.
In addition, the preparation method is simple, is suitable for large-scale production, is convenient to carry, has good stability and long storage time, can be directly brewed and drunk, can be used as an additive component for food processing, and has wide application.
Specifically, the present invention mainly relates to the following aspects.
In a first aspect, the invention provides a dendrobium nutrition powder containing a purified dendrobium officinale extract and capable of reducing blood sugar, wherein the dendrobium nutrition powder has a core-shell structure and at least comprises: amino acid modified slow-digestion starch of dendrobium officinale, purified polysaccharide of dendrobium officinale and dendrophenol.
The dendrobium officinale purified polysaccharide is a specific polysaccharide component purified by the method, and is beneficial to reducing blood sugar. The dendrophenol can be a commercially available food-grade pure dendrophenol product or a finished product prepared by the method.
In a second aspect, the invention provides a preparation method of the dendrobium officinale nutrient powder, which comprises the following preparation steps:
(1) extracting free amino acid from dendrobium officinale leaves;
(2) preparing modified slowly digestible starch by using the extracted amino acid of the dendrobium officinale;
(3) extracting and purifying a dendrobium officinale polysaccharide component, wherein the relative molecular weight of the polysaccharide component is more than 150 KDa;
(4) extracting and purifying dendrophenol from the dendrobium officinale degreasing waste liquid in the step of extracting polysaccharide;
(5) preparing slurry and constructing nutrient powder particles, wherein the mass ratio of the dendrobium candidum polysaccharide to the dendrophenol is 100: 0.1-0.5.
The preparation method of the nutritional powder comprises the following steps of (1): (a) sieving the washed, enzyme-deactivated, dried and crushed dendrobium officinale leaf powder with a 100-mesh sieve, adding distilled water with the mass being 10-40 times that of the powder, leaching the powder in a water bath oscillator at 50-60 ℃ for 20-30min, after leaching, carrying out vacuum filtration on the hot leaching solution, collecting filtrate, and cooling to room temperature; adding 0.5-1 times volume of anhydrous ethanol into the filtrate to remove residual protein, purifying at 4 deg.C for 6-18h, centrifuging at 8000-; the extraction content of the crude product of the free amino acid is 35-50 mg/g;
(b) dissolving the crude amino acid product with 6-8 times of 50-60% ethanol, concentrating, and vacuum evaporating for crystallization to obtain refined amino acid crystal with purity of above 99%.
The main amino acids in the obtained crystal are aspartic acid, glutamic acid, glycine, valine and leucine, which account for about half of the total amino acid weight.
Among them, preferably, the obtained amino acid crystals are further recrystallized and purified.
In the preparation method, the amino acid modified slow-digestion starch prepared in the step (2) is treated by a hydrothermal method without enzyme and acid-base additives, and the content of the slow-digestion starch is improved by further modification treatment on the basis of unchanged amino acid content, and the preparation method specifically comprises the following steps:
(a) dissolving slowly digestible starch in distilled water, and stirring at room temperature to prepare 20-40% slowly digestible starch emulsion;
(b) adding free amino acid crystals extracted from dendrobium officinale leaves into the slowly-digested starch milk, magnetically stirring at room temperature for 20-30min, and drying until the water content is 50-60%;
(c) putting the product obtained in the step into a closed heat-preservation reactor, and preserving heat for 10-16 h at 55-60 ℃;
(d) and taking out the reactant, naturally cooling to room temperature, drying in vacuum until the moisture content is less than 10%, crushing, and sieving to obtain the dendrobium officinale free amino acid modified slowly-digestible starch, wherein the content of the modified slowly-digestible starch is over 78%.
Wherein, the slow-digestion starch can be obtained commercially or by the common preparation method in the field, and the content of the slow-digestion starch is preferably more than 40 percent (calculated by the Englyst method).
The slowly digestible starch source can be any natural food material source in the field, such as corn slowly digestible starch, wheat slowly digestible starch or konjak slowly digestible starch.
Wherein, the proportion of the added amino acid to the slowly digestible starch in the steps is as follows: the mass ratio of the dry weight of the slowly digestible starch to the amino acid crystals is 20-30: 1.
In the preparation method, the specific steps of extracting and purifying the dendrobium officinale polysaccharide component in the step (3) are as follows:
(a) preparing a dendrobium polysaccharide crude product: sieving the dendrobium officinale stem micro powder with a 60-mesh sieve, soaking the dendrobium officinale stem micro powder in food-grade 95% ethanol with the mass of 8-10 times of that of the dendrobium officinale stem micro powder for 30min, and then performing reflux degreasing for 1-1.5 h; dissolving the degreased product powder with 15-20 times of distilled water, heating in water bath to 50-55 deg.C, adding cellulase 1-2% of the dry powder of herba Dendrobii, performing enzymolysis for 1-2 hr, destroying cell wall, and fully releasing polysaccharide component; heating the enzymolysis solution to 90-95 deg.C, inactivating enzyme at high temperature, maintaining the temperature for 30-60min, extracting with ultrasound for 1-2min every 5min, filtering, and collecting filtrate; adding 10 times of distilled water into the filter residue, heating to 90-95 deg.C, maintaining the temperature for 40-60min, ultrasonically extracting for 1-2min every 5min, filtering, mixing filtrates, vacuum concentrating the filtrate to one tenth volume by a rotary evaporator, adding 3-4 times of volume of anhydrous ethanol into the concentrated solution, standing at 0-4 deg.C or ice bath for 12-24 hr to precipitate polysaccharide, filtering, washing the precipitate with 75% edible ethanol, oven drying, and pulverizing to obtain crude polysaccharide product containing protein; dissolving the polysaccharide extract crude product in water of 20-50 times of the mass, adding Sevag reagent (isoamyl alcohol: chloroform is prepared at a ratio of 1: 4) of 20-25% in volume ratio, centrifuging for 15-20min at the normal temperature at 3000-4000r/min after ultrasonic oscillation for 3-5min, and taking supernatant to repeat the ultrasonic oscillation-centrifugation operation for 3-4 times until protein removal is complete; mixing the supernatants, adding 3-4 times volume of ethanol, standing in ice bath for 10-12 hr, filtering, collecting precipitate, washing with anhydrous ethanol, evaporating to remove solvent, and vacuum drying to obtain crude product of herba Dendrobii polysaccharide;
(b) purifying polysaccharide:
dissolving the above crude product of herba Dendrobii polysaccharide in water, dialyzing with dialysis membrane (molecular weight cutoff of 4000Da) for 24-48h to remove monosaccharide and oligosaccharide, taking out the liquid in dialysis bag, centrifuging at 10000-; separating and purifying the polysaccharide prefabricated product by using a Sephacryl S-200 (molecular sieve effect) propylene dextran column chromatography or a DEAE-52 cellulose ion exchange column eluted in a NaCl stage, collecting main peak components, combining and concentrating polysaccharide components (accounting for more than 73 percent of the total sugar) with the concentration of more than 150KDa, adding a proper amount of ethanol (till the ethanol content is 70-75 percent), placing for 12 hours in an ice bath, filtering, collecting precipitates, washing the precipitates with absolute ethanol, precipitating, and freeze-drying to obtain the dendrobium officinale polysaccharide refined product, wherein the yield of the polysaccharide refined product is 13.8 percent based on raw materials. The content of polysaccharide is above 95% by phenol-sulfuric acid method, and the content of protein is below 0.5% by bradford method.
Wherein, the polysaccharide crude product can also adopt a commercially available dendrobium officinale polysaccharide extract and is further purified by adopting the purification method of the invention.
In the preparation method of the invention, the specific steps of extracting and purifying the dendrophenol in the step (4) are as follows:
collecting ethanol extraction components discarded in the step of reflux degreasing in the preparation of dendrobe polysaccharide, concentrating under reduced pressure, extracting with n-hexane for 2-3 times, extracting with ethyl acetate for 3-4 times, and evaporating to remove filtrate to obtain crude extract of dendrophenol; adding appropriate amount of 20% sodium carbonate solution, stirring for 2-3 hr, standing for 4-12 hr, centrifuging at high speed, collecting supernatant, concentrating, charging pure carbon dioxide, acidifying for 1-2 hr to obtain white flocculent precipitate, centrifuging to obtain precipitate, and washing with distilled water twice; adding ethanol into the precipitate until the precipitate is completely dissolved, performing adsorption separation on a reverse phase silica gel column at 2-3BV/h, performing gradient elution by a water-ethanol system, performing tracking detection by thin-layer chromatography, collecting dendrophenol section eluent, performing reduced pressure concentration to obtain a dendrophenol crude product, crystallizing by absolute ethanol, and recrystallizing for 2-3 times to obtain a white dendrophenol crystal, wherein the extraction rate is 0.5mg/g based on the dry dendrobe powder.
Wherein the mass proportion of the dendrophenol crude extract to the 20% sodium carbonate solution is 1: 10-20.
In the preparation method, the specific steps of constructing the nutrient powder particles in the step (5) are as follows:
(a) adding the prepared dendrobe amino acid modified slow-digestion starch into distilled water to prepare 15-30 wt% of starch emulsion, uniformly stirring on a magnetic stirrer at a constant speed until the starch emulsion is uniform, adding trehalose, gellan gum and the dendrobe polysaccharide and dendrophenol prepared by the steps while stirring, uniformly stirring, and homogenizing by using a homogenizer, wherein the contents of the mixed dendrobe polysaccharide, dendrobe phenol, trehalose and gellan gum are as follows: 5-15 wt% of dendrobe polysaccharide, 2-3 wt% of trehalose, 0.1 wt% of gellan gum and 0.1-0.5 wt% of dendrobe phenol; carrying out electrostatic spray drying on the homogenized mixed solution to prepare gel particle powder;
(b) adding a proper amount of hydroxypropyl methyl cellulose, stevioside and chitosan into a 3-5 wt% sodium alginate solution, homogenizing and stirring uniformly to obtain viscous slurry, wherein the mass percentage of each raw material is as follows: 3-5 wt% of sodium alginate, 1-3 wt% of hydroxypropyl methylcellulose, 0.1-0.3 wt% of stevioside and 0.1-0.2 wt% of chitosan; and (3) uniformly spraying the viscous slurry on the surface of the gel particle powder by using a spraying dryer, so as to prepare the dendrobium nutritional powder particles.
Wherein the spraying dosage of the viscous slurry is one third to one half of the weight of the gel particle powder.
In a third aspect of the invention, the dendrobium nutrition powder prepared by the preparation method is provided.
In a fourth aspect of the invention, there is provided the use of a nutritional powder, either as a reconstituted product for direct consumption or as an additive in food processing, in the processing of a staple food.
In a fifth aspect of the invention, a food, a health product or a pharmaceutical composition is provided, wherein the food, the health product or the pharmaceutical composition comprises the dendrobium nutritional powder particles.
In the nutritional powder of the present invention, references to "comprising" of composition encompass both open "comprising" and "comprises" and the like, and also closed "consisting of … and the like, and the like.
The benefits of the present invention also include, but are not limited to, the following:
compared with the dendrobium micropowder or polysaccharide crude product composition in the prior art, unnecessary monosaccharide/oligosaccharide such as glucose and the like, alkaloid with the effect of increasing blood sugar and other useless components are removed, the beneficial polysaccharide components are purified, concentrated and screened, the dendrobium phenol component used for improving diabetic complications and the dendrobium amino acid modified high-content slow-digestion starch are supplemented in a reasonable proportion, the dendrobium raw material is fully and efficiently utilized, the purpose of synergistic effect improvement on blood sugar is achieved, the dendrobium micropowder or polysaccharide crude product composition is not only suitable for the hyperglycemia crowds to take daily for reducing blood sugar, but also suitable for the ordinary crowds to take daily for preventing diabetes, obesity and the like due to the fact that polysaccharide, particularly dendrobium phenol, does not stimulate insulin secretion.
In addition, the preparation method is simple, is suitable for large-scale production, and has good application prospect and market value.
Detailed Description
The present invention is described in detail below with reference to specific preparation examples and examples, but the use and purpose of these exemplary embodiments are merely to illustrate the present invention, and do not constitute any limitation to the actual scope of the present invention in any form, and the scope of the present invention is not limited thereto.
Preparation example 1: extraction of free amino acids from dendrobium officinale leaves
(1) 50 g of the washed, enzyme-deactivated, dried and crushed dendrobium officinale leaf powder is sieved by a 100-mesh sieve, 1500ml of distilled water is added, the mixture is leached for 30min at 60 ℃ in a water bath oscillator, after the leaching is finished, the leaching solution is subjected to vacuum filtration while being hot, 30ml of filtrate is collected and cooled to room temperature;
(2) adding 0.5 times volume of absolute ethyl alcohol into the filtrate to remove residual protein, purifying at 4 ℃ for 12h, centrifuging at 8000r/min for 20min, collecting supernatant, filtering with nanofiltration membrane with molecular weight cutoff of 1000Da, concentrating the filtrate under reduced pressure, and vacuum drying at 50 ℃ to obtain 2.5 g of crude amino acid; the extraction content of the crude product of the free amino acid is 50 mg/g;
(3) dissolving the crude amino acid product with 20ml of 50% ethanol, concentrating, and vacuum evaporating for crystallization to obtain refined amino acid crystal with purity of above 99%.
Preparation example 2: preparation of amino acid modified slow-digestion starch of dendrobium officinale
(1) Dissolving 100g of konjac slowly digestible starch in distilled water, and stirring at room temperature to prepare 20-40% of slowly digestible starch emulsion; adding 5 g of the free amino acid crystals extracted from the dendrobium officinale leaves into the slowly digested starch milk, magnetically stirring the mixture at room temperature for 30min, and drying the mixture until the water content is 50%;
(2) putting the product obtained in the step into a closed heat-preservation reactor, and preserving heat for 16 hours at the temperature of 55 ℃;
(3) and taking out the reactant, naturally cooling to room temperature, drying in vacuum until the moisture content is less than 10%, crushing, and sieving to obtain the dendrobium officinale free amino acid modified slowly-digestible starch, wherein the content of the modified slowly-digestible starch is about 82% measured by an Englyst method.
Preparation example 3: extracting and purifying dendrobium officinale polysaccharide component
(1) Preparing a dendrobium polysaccharide crude product: weighing 100g of clean dendrobium officinale stem micro powder, sieving the powder with a 60-mesh sieve, soaking the powder in 1000ml of food-grade 95% ethanol for 30min, and then carrying out reflux degreasing for 1.5 h; dissolving the dried degreased product powder with distilled water 15 times the mass of the degreased product powder, heating the degreased product powder to 55 ℃ in a water bath, adding cellulase 2% of the mass of the dendrobium dry powder for enzymolysis for 2 hours, breaking cell walls, and fully releasing polysaccharide components; heating the enzymolysis solution to 95 deg.C, inactivating enzyme at high temperature, maintaining the temperature for 60min, extracting with ultrasound for 1min every 5min, filtering, and collecting filtrate; adding 10 times of distilled water into the filter residue, heating to 90 ℃, keeping the temperature for 40min, carrying out ultrasonic extraction for 2min every 5min, filtering, combining the filtrates, carrying out vacuum concentration on the filtrate to one tenth of the volume by using a rotary evaporator, adding 3 times of volume of absolute ethyl alcohol into the concentrated solution, standing for 18 hours at 0-4 ℃ or in an ice bath to precipitate polysaccharide, filtering, washing the precipitate by using 75% edible-grade ethanol, drying and crushing the precipitate to obtain a crude polysaccharide product containing protein;
(2) dissolving the polysaccharide extract crude product in 20 times of water by mass, adding 20% volume ratio of Sevag reagent (isoamylol: chloroform: 1: 4), centrifuging at 4000r/min at normal temperature for 20min after ultrasonic oscillation for 5min, and repeating the ultrasonic oscillation-centrifugation operation for 3 times to remove protein completely; mixing the supernatants, adding 4 times volume of ethanol, standing in ice bath for 12 hr, filtering, collecting precipitate, washing with anhydrous ethanol, evaporating to remove solvent, and vacuum drying to obtain crude product of herba Dendrobii polysaccharide;
(3) purifying polysaccharide:
dissolving the protein-removed dendrobe polysaccharide crude product in water, dialyzing with a dialysis membrane with molecular weight cutoff of 4000Da for 24h to remove monosaccharide and oligosaccharide, taking out liquid in a dialysis bag, centrifuging at 12000rpm for 15min, concentrating supernatant, and freeze-drying to obtain dendrobe polysaccharide prefabricated product;
(4) separating and purifying the polysaccharide prefabricated product by using a molecular sieve effect propylene dextran column chromatography (Sephacryl S-200) or a DEAE-52 cellulose ion exchange column eluted in a NaCl stage, collecting main peak components, combining and concentrating polysaccharide components with the concentration of more than 150KDa, adding a proper amount of ethanol until the ethanol content is 70%, placing in an ice bath for 12 hours, filtering, collecting precipitates, washing the precipitates with absolute ethanol, precipitating, and freeze-drying to obtain the dendrobium officinale polysaccharide refined product, wherein the yield of the dendrobium officinale polysaccharide refined product is 13.8% calculated by raw materials. The content of polysaccharide is 97% by phenol-sulfuric acid method, and the content of protein is 0.4% by bradford method.
Preparation example 4: extracting and purifying dendrophenol
Collecting the ethanol extraction solution obtained in the reflux degreasing step in the preparation example 3, concentrating to 50ml under reduced pressure, extracting with n-hexane for 2-3 times, extracting with ethyl acetate for 3-4 times, and evaporating to remove the filtrate to obtain 5 g of dendrophenol crude extract; adding 50 g of 20% sodium carbonate solution, stirring and reacting for 3 hours, standing for 12 hours, centrifuging at a high speed of 6000r/min, collecting supernatant, concentrating, charging pure carbon dioxide, acidifying for 1 hour to obtain white flocculent precipitate, centrifuging, taking the precipitate, and washing twice with distilled water; adding ethanol into the precipitate until the precipitate is completely dissolved, performing adsorption separation on a reverse phase silica gel column at 3BV/h, performing gradient elution by a water-ethanol system, performing tracking detection by a thin-layer chromatography, collecting dendrophenol section eluent, performing reduced pressure concentration to obtain a dendrophenol crude product, crystallizing by absolute ethanol, and recrystallizing for 2 times to obtain a white dendrophenol crystal of 50.3mg, wherein the purity is 99%, and the extraction rate is 0.5mg/g calculated by dry dendrobe powder.
Preparation example 5: construction of nutrient powder particles
(1) Adding 100g of prepared dendrobe amino acid modified slow-digestion starch into distilled water to prepare 500 g of 20 wt% starch emulsion, uniformly stirring the mixture on a magnetic stirrer at a constant speed until the mixture is uniform, adding a proper amount of trehalose, gellan gum and dendrobe polysaccharide and dendrobe phenol prepared by the steps while stirring, and homogenizing the mixture by a homogenizer after the mixture is uniformly stirred, wherein the contents of the mixed dendrobe polysaccharide, dendrobe phenol, trehalose and gellan gum are as follows: 10 wt% of dendrobe polysaccharide, 2 wt% of trehalose, 0.1 wt% of gellan gum and 0.01 wt% of dendrophenol; carrying out electrostatic spray drying on the homogenized mixed solution to prepare gel particle powder;
(2) adding a proper amount of hydroxypropyl methyl cellulose, stevioside and chitosan into a 3 wt% sodium alginate solution, homogenizing and stirring uniformly to obtain 500 g of viscous slurry, wherein the mass percentage of each raw material is as follows: 3 wt% of sodium alginate, 1 wt% of hydroxypropyl methylcellulose, 0.1 wt% of stevioside and 0.2 wt% of chitosan; and (3) uniformly spraying the viscous slurry on the surface of the rapidly-rolled gel particle powder by using a spraying dryer, and carrying out vacuum drying and sterilization to prepare the dendrobium nutritional powder particles.
Comparative example 1: extracting and purifying 3-10KDa dendrobium officinale polysaccharide component
Comparative example 1 was performed in the same manner as in preparation example 3, except that the polysaccharide fraction of Dendrobium officinale was purified and collected in the range of 3-10 kDa.
Comparative example 2: extracting and purifying 10-150KDa dendrobium officinale polysaccharide component
Comparative example 2 was performed according to the same method as in preparation example 3, except that the polysaccharide fraction of Dendrobium officinale ranging from 10 to 150kDa was purified and collected.
Effect example 1: hypoglycemic testing
The test method comprises the following steps: healthy male KM mice were selected, weighing 20g (up-down difference rate 10%), and randomly divided into 5 groups: blank control group, model control group and 3 test groups; wherein the polysaccharide components administered to the test group are: preparation example 3 polysaccharide fraction, comparative example 1 polysaccharide fraction, comparative example 2 polysaccharide fraction, 10 animals per group of experimental animals, and the experimental period was 30 days.
Establishing a mouse diabetes model: the mice are fasted for 12 hours, the disposable intraperitoneal injection of streptozotocin is 15mg/100g of body weight, the mice are fasted for 12 hours after the modeling for one week, and the model with the measured fasting blood sugar value higher than 11mmol/L is the diabetes model with successful modeling.
Wherein, blank control group: the normal mice are infused with equivalent distilled water every day;
model control group: the diabetic mice are infused with equivalent distilled water every day;
test group: the diabetic mice were gavaged daily with 120mg/kg.
The values are averaged and the experimental results are as follows:
TABLE 1
As can be seen from the above table, the polysaccharide fraction (greater than 150KDa) in preparation example 3 significantly reduced the fasting blood glucose level and the area under the blood glucose curve in diabetic mice, compared to the model control group; the polysaccharide fraction (10-150kDa) of comparative example 2 reduced the fasting blood glucose level in diabetic mice to a certain extent; the polysaccharide fraction (3-10kDa) in comparative example 1 was only slightly effective in lowering the fasting blood glucose level in diabetic mice.
The experimental result shows that compared with low molecular weight polysaccharide, the dendrobium polysaccharide component with the molecular weight of more than 150KDa has obvious blood sugar reducing effect on diabetic mice. Therefore, the nutrition powder particles have the effect of reducing blood sugar for diabetes and can improve diabetic complications.
It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should also be understood that various alterations, modifications and/or variations can be made to the present invention by those skilled in the art after reading the technical content of the present invention, and all such equivalents fall within the protective scope defined by the claims of the present application.
Claims (2)
1. A preparation method of dendrobium nutrition powder containing a purified dendrobium officinale extract comprises the following steps:
(1) extracting free amino acid from dendrobium officinale leaves;
(2) preparing modified slowly digestible starch by using the extracted free amino acid of the dendrobium officinale;
(3) extracting and purifying a dendrobium officinale polysaccharide component, wherein the relative molecular weight of the polysaccharide component is more than 150 KDa;
(4) extracting and purifying dendrophenol from the dendrobium officinale degreasing fluid in the step of extracting polysaccharide;
(5) preparing slurry and constructing dendrobium nutrition powder particles, wherein the mass ratio of the dendrobium candidum polysaccharide to the dendrophenol is 100: 0.1-0.5;
wherein, the step (1) is specifically prepared as follows:
(a) sieving the washed, enzyme-deactivated, dried and crushed dendrobium officinale leaf powder with a 100-mesh sieve, adding distilled water with the mass being 10-40 times that of the powder, leaching the powder in a water bath oscillator at 50-60 ℃ for 20-30min, after leaching, carrying out vacuum filtration on the hot leaching solution, collecting filtrate, and cooling to room temperature; adding 0.5-1 times volume of anhydrous ethanol into the filtrate, purifying at 4 deg.C for 6-18h to remove residual protein, centrifuging at 8000-; the extraction content of the crude product of the free amino acid is 35-50 mg/g;
(b) dissolving the crude amino acid product in 50-60% ethanol 6-8 times the mass of the crude amino acid product, concentrating, and vacuum evaporating for crystallization to obtain refined amino acid crystal;
wherein the specific preparation process of the step (2) is as follows:
1) dissolving slowly digestible starch in distilled water, and stirring at room temperature to prepare a slowly digestible starch emulsion with the mass fraction of 20-40%;
2) adding free amino acid crystals extracted from dendrobium officinale leaves into the slowly digestible starch emulsion, magnetically stirring at room temperature for 20-30min, and drying until the water content is 50-60%;
3) putting the product obtained in the step into a closed heat-preservation reactor, and preserving heat for 10-16 h at 55-60 ℃;
4) taking out the reactant, naturally cooling to room temperature, vacuum drying until the moisture content is less than 10%, crushing, and sieving to obtain the dendrobium officinale free amino acid modified slowly digestible starch;
wherein, the step (3) is specifically prepared as follows:
1) preparing a dendrobium polysaccharide crude product: sieving the dendrobium officinale stem dry powder with a 60-mesh sieve, soaking the dendrobium officinale stem dry powder in food-grade 95% ethanol with the mass of 8-10 times of that of the dendrobium officinale stem dry powder for 30min, and then performing reflux degreasing for 1-1.5 h; dissolving the degreased product powder with 15-20 times of distilled water, heating in water bath to 50-55 deg.C, adding cellulase 1-2% of the dry powder of caulis Dendrobii stem, performing enzymolysis for 1-2 hr, breaking cell wall, and fully releasing polysaccharide component; heating the enzymolysis solution to 90-95 deg.C for inactivating enzyme at high temperature, maintaining the temperature for 30-60min, extracting with ultrasound for 1-2min every 5min, filtering, and collecting filtrate; adding 10 times of distilled water into the filter residue, heating to 90-95 deg.C, maintaining the temperature for 40-60min, ultrasonically extracting for 1-2min every 5min, filtering, mixing filtrates, vacuum concentrating the filtrate to one tenth volume by a rotary evaporator, adding 3-4 times of volume of anhydrous ethanol into the concentrated solution, standing at 0-4 deg.C or ice bath for 12-24 hr to precipitate polysaccharide, filtering, washing the precipitate with 75% edible ethanol, oven drying, and pulverizing to obtain crude polysaccharide product containing protein; dissolving the crude polysaccharide extract product in water with the mass of 20-50 times, and adding a Sevag reagent with the volume ratio of 20-25%, wherein the Sevag reagent is prepared from isoamylol: chloroform is 1: 4, centrifuging for 15-20min at 3000-; adding 3-4 times volume of ethanol into the supernatant, standing in ice bath for 10-12 hours, filtering, collecting precipitate, washing with absolute ethanol, evaporating to remove solvent, and vacuum drying to obtain crude product of herba Dendrobii polysaccharide;
2) purifying polysaccharide: dissolving the crude product of dendrobe polysaccharide in water, dialyzing with a dialysis membrane with molecular weight cutoff of 4000Da for 24-48h to remove monosaccharide and oligosaccharide, taking out the liquid in the dialysis bag, centrifuging at 10000-12000rpm for 10-15min, concentrating the supernatant, and freeze-drying to obtain a dendrobe polysaccharide prefabricated product; separating and purifying the polysaccharide prefabricated product by using a propylene dextran column chromatography with a molecular sieve effect or a DEAE-52 cellulose ion exchange column eluted in a NaCl stage, collecting main peak components, combining and concentrating polysaccharide components with the concentration of more than 150KDa, adding a proper amount of ethanol, standing in an ice bath for 12 hours, filtering, collecting precipitates, washing the precipitates with absolute ethanol, precipitating, and freeze-drying to obtain the dendrobium officinale polysaccharide refined product;
wherein, the step (4) is specifically prepared as follows:
collecting ethanol extraction components in the reflux degreasing step in the preparation of dendrobe polysaccharide, concentrating under reduced pressure, extracting with n-hexane for 2-3 times, extracting with ethyl acetate for 3-4 times, and evaporating to remove filtrate to obtain crude extract of dendrophenol; adding 20% sodium carbonate solution, stirring for 2-3 hr, standing for 4-12 hr, centrifuging at high speed, collecting supernatant, concentrating, charging pure carbon dioxide, acidifying for 1-2 hr to obtain white flocculent precipitate, centrifuging to obtain precipitate, and washing with distilled water twice; adding ethanol into the precipitate until the precipitate is completely dissolved, performing adsorption separation on a reverse phase silica gel column at 2-3BV/h, performing gradient elution by a water-ethanol system, performing tracking detection by thin-layer chromatography, collecting dendrophenol section eluate, concentrating under reduced pressure to obtain a dendrophenol crude product, crystallizing by absolute ethanol, and recrystallizing for 2-3 times to obtain a white dendrophenol crystal; wherein the mass proportion of the dendrophenol crude extract to the sodium carbonate solution is 1: 10-20 parts of;
wherein, the step (5) is specifically prepared as follows:
1) adding the prepared dendrobe amino acid modified slow-digestion starch into distilled water to prepare 15-30 wt% of starch emulsion, uniformly stirring on a magnetic stirrer at a constant speed until the starch emulsion is uniform, adding trehalose, gellan gum and the dendrobe polysaccharide and dendrophenol prepared by the steps while stirring, uniformly stirring, and homogenizing by using a homogenizer, wherein the contents of the mixed dendrobe polysaccharide, dendrobe phenol, trehalose and gellan gum are as follows: 5-15 wt% of dendrobe polysaccharide, 2-3 wt% of trehalose, 0.1 wt% of gellan gum and 0.1-0.5 wt% of dendrophenol; carrying out electrostatic spray drying on the homogenized mixed solution to prepare gel particle powder;
2) adding hydroxypropyl methyl cellulose, stevioside and chitosan into a 3-5 wt% sodium alginate solution, homogenizing and stirring uniformly to obtain viscous slurry, wherein the mass percentage of each raw material is as follows: 3-5 wt% of sodium alginate, 1-3 wt% of hydroxypropyl methylcellulose, 0.1-0.3 wt% of stevioside and 0.1-0.2 wt% of chitosan; and (3) uniformly spraying the viscous slurry on the surface of the gel particle powder by using a spraying dryer, so as to prepare the dendrobium nutritional powder containing the purified dendrobium officinale extract.
2. The dendrobium nutrition powder containing the purified dendrobium officinale extract prepared by the preparation method of claim 1.
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