CN114099561B - Dandelion flower polysaccharide extract with antibacterial and probiotic activities and preparation method thereof - Google Patents
Dandelion flower polysaccharide extract with antibacterial and probiotic activities and preparation method thereof Download PDFInfo
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- CN114099561B CN114099561B CN202111147649.2A CN202111147649A CN114099561B CN 114099561 B CN114099561 B CN 114099561B CN 202111147649 A CN202111147649 A CN 202111147649A CN 114099561 B CN114099561 B CN 114099561B
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- SZDMSNWMQAMVTJ-UHFFFAOYSA-N quercetin-3-O-glucoside Natural products OC1OC(COC2=C(C(=O)c3cc(O)cc(O)c3O2)c4ccc(O)c(O)c4)C(O)C(O)C1O SZDMSNWMQAMVTJ-UHFFFAOYSA-N 0.000 description 1
- FZKBNCDAGYDHTP-UHFFFAOYSA-N quercetin-3-O-glycoside Natural products OC1C(O)C(O)C(O)OC1COC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O FZKBNCDAGYDHTP-UHFFFAOYSA-N 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- 235000005493 rutin Nutrition 0.000 description 1
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 1
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 1
- 229960004555 rutoside Drugs 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 229940005605 valeric acid Drugs 0.000 description 1
Images
Classifications
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/288—Taraxacum (dandelion)
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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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A61P31/04—Antibacterial agents
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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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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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A61K2236/50—Methods involving additional extraction steps
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- A61K2800/10—General cosmetic use
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a dandelion flower polysaccharide extract with antibacterial and probiotic activities and a method, which comprises the following steps: the dandelion flower polysaccharide extract is prepared by the steps of taking fresh dandelion flowers as raw materials, drying the fresh dandelion flowers in the sun, crushing the fresh dandelion flowers (the moisture content is lower than 4%), taking distilled water as a solvent, stirring the distilled water at a high temperature to carry out boiling extraction on active substances in the dandelion flowers, carrying out vacuum filtration while the dandelion flowers are hot, collecting filtrate, carrying out rotary evaporation, concentration and cooling the filtrate to room temperature, adding a proper amount of absolute ethyl alcohol and a concentrated solution according to a certain volume ratio, carrying out alcohol precipitation at a proper temperature for a period of time, carrying out vacuum filtration, collecting alcohol precipitates, and carrying out vacuum freeze drying or spray drying on the alcohol precipitates. The dandelion flower polysaccharide extract has the characteristics of low equipment investment, simple operation, short time, high efficiency, high quality, suitability for industrialization and the like. Can provide a new idea for high-valued intensive processing of dandelion flowers in China.
Description
Technical Field
The invention relates to the field of natural product chemistry for preparing fine chemicals with high added values by purifying and enriching chemical active ingredients of natural products, in particular to a dandelion flower polysaccharide extract with antibacterial and probiotic activities and a method thereof.
Background
Dandelion was originally recorded in Tang Dynasty "compendium of materia Medica" as Taraxacuum Webar of Dang Ding Dang Ying genus Taraxacum, about more than 2000 species worldwide. There are 70 varieties of Chinese, 1 variety. Taraxacum Mongolon (TM) is a perennial herb of Compositae, which is homologous in medicine and food, is listed in the list of food and medicine by Ministry of health, and has the effects of clearing heat and removing toxicity, relieving swelling and resolving masses, inducing diuresis and treating stranguria, invigorating stomach and diminishing inflammation, protecting liver and benefiting gallbladder and the like.
Dandelion flowers are a by-product of dandelion planting industry, and contain many other bioactive substances, such as polysaccharides, phenolic acids, flavonoids and the like. The flavonoids include luteolin, quercetin, apigenin, rutin and their glycoside derivatives, such as luteolin-7-beta-D-glucoside, and quercetin-3-O-glucoside. The phenolic acid in the dandelion flower comprises chlorogenic acid, caffeic acid and the like, and has the effects of resisting inflammation, bacteria, viruses, tumors and the like. Regarding polysaccharide enrichment and extraction and separation of flavone and phenolic acid in dandelion flowers, the patent application at home and abroad finds that foreign patents have fewer reports on dandelion flowers, especially water-soluble active polysaccharide extracts, and the foreign patent application number is US201113068283, which describes a method for extracting rubber and carbohydrate from dandelion roots, and is significantly different from the target products dandelion flower polysaccharide and phenolic acid related in the invention. Domestic applications relate to patents of polysaccharides in dandelion flowers including: a herba Taraxaci extract and its preparation method, patent application No: CN201110062814.4 and CN201210521873.8, in the process of extracting flavone, 65% alcohol-water extraction is used, petroleum ether is used for decoloring, organic solvent residue exists, inorganic strong acid (hydrochloric acid) or acetic acid is added, corrosion of a metal extraction container is easily caused in the process of extraction, and the production cost is improved, so that the production benefit of enterprises is not improved. In addition, the raw material used in the patent is the whole dandelion, and the raw material used in the invention is dandelion flowers, so that the active substances can be directionally and accurately obtained, and the introduction of other impurities is reduced. The patent application number is CN02111738.1, and when the dandelion preparation is prepared, water extract and pharmaceutic adjuvant are directly prepared to be compounded into the dandelion preparation which is mainly used for enriching organic acid of the dandelion. A herba Taraxaci extract is disclosed, and its preparation method is patent No. 201910729006.5, and the obtained herba Taraxaci extract is mainly used for obtaining herba Taraxaci flavone and herba Taraxaci pigment as natural colorant.
In recent years, studies have shown that plant polysaccharides exert various functional activities and may be closely related to the regulation of intestinal flora. Because the organism lacks polysaccharide hydrolase, most of polysaccharide ingested by oral administration can not be directly digested and absorbed by human body, but intestinal flora in the body plays a role in the following two aspects, 1) the intestinal flora can degrade the polysaccharide to generate monosaccharide, the absorption and utilization of the polysaccharide by the organism are promoted, functional metabolites can be generated, such as short chain fatty acid, and 2) the polysaccharide can directly increase the number of beneficial bacteria and reduce the number of harmful bacteria by adjusting the proportion of the intestinal flora, so that the physiological level of the organism is improved. The intestinal flora decomposes polysaccharide chains to obtain energy by secreting glycoside hydrolase, lyase, esterase and the like, wherein the polysaccharide is glycolyzed into short-chain fatty acids such as formic acid, acetic acid, propionic acid, butyric acid, valeric acid and the like which are end products, and the short-chain fatty acids not only participate in the energy metabolism of animals, but also can be used as food of the intestinal flora and have good biological activity. Meanwhile, the type and the content of the short-chain fatty acid are directly related to the type and the composition proportion of intestinal flora. In addition, the short-chain fatty acid can also reduce the pH value of the intestinal tract, inhibit the multiplication of part of harmful colonies, improve the immunologic function of the intestinal mucosa and the like. This may be due to the relatively higher efficiency of growth of lactic acid bacteria, bifidobacteria using such substrates than other microorganisms, and the tolerance to short chain fatty acids and acidification microenvironment caused by fermentation.
Disclosure of Invention
The invention aims to fully and effectively utilize dandelion flowers, extract the active ingredient compound of polysaccharide, flavone and phenolic acid in the dandelion flowers and provide support for deep processing and development of the dandelion flowers.
The technical scheme adopted by the invention comprises the following steps: a taraxacum flower polysaccharide extract with antibacterial and probiotic activities and its preparation method are provided, wherein taraxacum flower is used as raw material, and water extraction and alcohol precipitation method is adopted to obtain taraxacum flower polysaccharide extract, which mainly contains polysaccharide, and is rich in flavone such as luteolin, quercetin, luteolin and organic acid, wherein the organic acid comprises p-coumaric acid, caffeic acid and chlorogenic acid; the dandelion flower polysaccharide extract with high-efficiency antibacterial and probiotic activities has an inhibiting effect on one or more of staphylococcus aureus (S.aureus), shigella (S.Castellani), escherichia coli (E.coli) and salmonella (S.enteritidis); has proliferation effect on one or both of Lactobacillus (L.Bulgaricus) and Bifidobacterium (Bifidobacterium).
The dandelion flower polysaccharide extract is obtained by taking dandelion flowers as a raw material and adopting a water extraction and alcohol precipitation method.
The dandelion flower is fresh Taraxacum mongolicum hand-held-Mazz produced by Sixuan, is picked and dried in the sun for use, and has a water content of less than 4%.
The dandelion flowers are crushed, water is used as a solvent, the mixture is stirred uniformly under a boiling condition, the hot filtrate is collected by vacuum filtration while the mixture is hot, the mixture is subjected to rotary evaporation, concentration and cooling to room temperature, absolute ethyl alcohol and concentrated solution are added for alcohol precipitation, and after vacuum filtration, alcohol precipitates are collected and then subjected to vacuum freeze drying or water adding and redissolving, and spray drying is carried out to prepare the dandelion flower polysaccharide extract.
The boiling condition refers to that water with the temperature of 100 ℃ is distilled water or deionized water, the solid-liquid ratio of dandelion pollen to water ranges from 1:1 to 60g/mL, the mechanical stirring speed is 10 to 1000r/min, and the stirring time is 1 to 240min.
The rotary evaporation concentration condition is that the temperature is 45-75 ℃, and the final concentration volume range is 200-300 mL.
The volume ratio of the absolute ethyl alcohol to the concentrated solution is 1:1-10, and the temperature range of alcohol precipitation is-4 ℃.
The time range of the vacuum freeze drying is 36-72 h, the vacuum degree pressure range is 80-110 Pa, or the time range of the spray drying is 1-3 h, and the vacuum degree pressure range is 0.09-0.1M Pa.
Has the advantages that:
(1) The dandelion flower polysaccharide extract prepared by the invention is rich in organic acids such as chlorogenic acid, caffeic acid and flavone compounds; (2) Has potent antibacterial activity such as Staphylococcus aureus (S.aureus), shigella (S.Castellani), escherichia coli (E.coli) and Salmonella (S.Enteritidis) and probiotic activity such as proliferation of Lactobacillus (L.Bulgaricus) and Bifidobacterium (Bifidobacterium).
(2) The invention adopts a water extraction and alcohol precipitation method, has no organic solvent residue, can reduce the investment cost compared with an organic solvent and strong acid and strong alkali, and provides a safe and high-quality auxiliary agent for medicines, cosmetics and health care products.
(4) The method has the advantages of simple and stable process, high product yield, good result reproducibility, good product quality, high investment benefit and easy realization of industrial production.
(5) The polysaccharide extract of taraxacum flower has stronger probiotic proliferation capability than the polysaccharide extracts of taraxacum leaves and whole plants.
Drawings
FIG. 1 is a flow chart of the processing technique of dandelion flower polysaccharide extract of the present invention.
FIG. 2 HPLC chart in Dandelion flower polysaccharide extract.
FIG. 3 shows the antibacterial effect of taraxacum flower polysaccharide extract.
Figure 4 the prebiotic effect of taraxacum flower polysaccharide extract.
Figure 5 comparison of the probiotic activity of dandelion flower polysaccharide extract compared to dandelion leaf polysaccharide extract and whole dandelion polysaccharide extract.
Detailed Description
A dandelion flower polysaccharide extract with antibacterial and probiotic activities and a method thereof, wherein the dandelion flower is prepared by picking fresh Taraxacum mongolicum hand-Mazz produced by Siyang Jiangsu, sun-drying, pulverizing to 60-mesh pollen serving as a raw material (the water content is less than 4%), taking distilled water as a solvent, mechanically stirring for a period of time according to a certain solid-liquid ratio under a boiling condition, carrying out vacuum filtration for collecting filtrate while heating, carrying out rotary evaporation, concentration and cooling to room temperature, adding a proper amount of absolute ethyl alcohol and a concentrated solution according to a certain volume ratio, carrying out alcohol precipitation for a period of time at a proper temperature, carrying out vacuum filtration, collecting alcohol precipitate, carrying out vacuum freeze drying or adding a proper amount of water for redissolving, and carrying out spray drying. The taraxacum polysaccharide extract has high effect in inhibiting Staphylococcus aureus (S.Aureaus), shigella (S.Castellani), escherichia coli (E.Coli) and Salmonella (S.Enteritidis), and has proliferation effect of Lactobacillus (L.Bulgaricus) and Bifidobacterium (Bifidobacterium).
The boiling condition refers to that water with the temperature of 100 ℃ is distilled water or deionized water, the solid-liquid ratio of dandelion pollen to water is 1:1-60 (g/mL), the mechanical stirring speed is 10-1000 r/min, and the stirring time is 1-240 min.
The rotary evaporation concentration condition is that the temperature is 45-75 ℃, and the final concentration volume range is 200-300 mL.
The volume ratio of the absolute ethyl alcohol to the concentrated solution is 1:1-1
The time range of the vacuum freeze drying is 36-72 h, the vacuum degree pressure range is 80-110 Pa or the spray drying time range is 1-3 h, the vacuum degree pressure range is 0.09-0.1M Pa.
The prepared dandelion flower polysaccharide extract is rich in flavones such as luteolin, quercetin, luteolin and the like besides polysaccharide, organic acids such as p-coumaric acid, caffeic acid, chlorogenic acid and the like, and the high-efficiency bacteriostasis refers to one or more of staphylococcus aureus (S.aureus), shigella (S.castellani), escherichia coli (E.coli) and salmonella (S.enteritidis). By highly effective probiotic activity is meant a significant proliferative effect on one or both of lactobacilli (l.bulgaricus) and bifidobacteria (bifidobacteria).
Example 1
Weighing 100g of dandelion flower dry powder, adding 3000mL of distilled water into a big beaker, extracting for 4h at a mechanical stirring speed of 1000r/min under the boiling condition of 100 ℃, repeatedly extracting for 2 times, carrying out suction filtration while hot, collecting filtrate for 2 times, carrying out rotary evaporation and concentration at 55 ℃ until the concentrated solution is about 300mL, cooling the concentrated solution to room temperature, adding 800mL of ethanol to precipitate out polysaccharide, placing the polysaccharide at the temperature of-4 ℃ for standing for 24h, carrying out vacuum suction filtration, and carrying out vacuum freeze drying (drying for 48h, the vacuum pressure range is 100 Pa) to obtain the solid content of the dandelion flower polysaccharide extract of 1.86g, wherein the solid content of the dandelion flower polysaccharide extract can reach 60.39%, the contents of chlorogenic acid, caffeic acid and p-coumaric acid are respectively 1.36%, 3.76% and 9.88%, and the total flavone is about 8.7%. When the concentration of the dandelion flower polysaccharide extract is 1.0mg/mL, the bacteriostatic efficiency of shigella and salmonella is 60% and 40% respectively, and the proliferation efficiency of probiotics exceeds 15%, such as the proliferation efficiency of lactobacillus and bifidobacterium is 17.5% and 15.2% respectively.
Example 2
Weighing 125g of dandelion flower dry powder, adding 3750mL of distilled water into a big beaker, extracting for 4h at a mechanical stirring rate of 700r/min under the boiling condition of 100 ℃, repeatedly extracting for 2 times, carrying out suction filtration while hot, collecting filtrate for 2 times, carrying out rotary evaporation and concentration at 60 ℃ until the concentrated solution is about 400mL, cooling the concentrated solution to room temperature, adding 1600mL of ethanol to precipitate polysaccharide, placing the polysaccharide at the temperature of-2 ℃ for standing for 24h, carrying out vacuum suction filtration, and carrying out vacuum freeze drying (drying for 60h, the vacuum pressure range is 90 Pa) to obtain the dandelion flower polysaccharide extract with the solid content of 2.23g, wherein the content of the total polysaccharide can reach 55.43%, the contents of chlorogenic acid, caffeic acid and p-coumaric acid are respectively 1.51%, 4.05% and 10.13%, and the total flavone is about 9.12%. When the concentration of the dandelion flower polysaccharide extract is 1.0mg/mL, the bacteriostatic efficiency of shigella and salmonella is 61.13% and 39.28% respectively, and the proliferation efficiency of probiotics such as lactobacillus and bifidobacterium is 19.3% and 16.8% respectively.
Example 3
Weighing 200g of dandelion flower dry powder, adding 6000mL of distilled water into a big beaker, extracting for 4h at a mechanical stirring speed of 800r/min under the boiling condition of 100 ℃, repeatedly extracting for 2 times, carrying out suction filtration while hot, collecting filtrate for 2 times, carrying out rotary evaporation and concentration at 50 ℃ until the concentrated solution is about 520mL, cooling the concentrated solution to room temperature, adding 2100mL of ethanol to precipitate out polysaccharide, standing for 24h at 0 ℃, carrying out vacuum filtration, and carrying out vacuum freeze drying (drying for 72h, the vacuum degree pressure range is 100 Pa) to obtain the dandelion flower polysaccharide extract with the solid content of 3.36g, wherein the dandelion flower polysaccharide extract contains 58.64% of total polysaccharide, 1.39%, 3.96% and 12.06% of chlorogenic acid, caffeic acid and p-coumaric acid respectively, and the total flavone is about 10.08%. When the concentration of the dandelion flower polysaccharide extract is 10.0mg/mL, the bacteriostatic efficiency of staphylococcus aureus and escherichia coli is 90.52% and 62.34% respectively, and the proliferation efficiency of probiotics such as lactobacillus and bifidobacterium is-10.1% and-12.5% respectively.
Example 4
Weighing 150g of dandelion flower dry powder, adding 4500mL of distilled water into a glass extractor, extracting for 3.5h at a mechanical stirring speed of 900r/min under the boiling condition of 100 ℃, repeatedly extracting for 2 times, carrying out suction filtration while hot, collecting 2 times of filtrate, carrying out rotary evaporation and concentration at 60 ℃ until the concentrated solution is about 350mL, cooling the concentrated solution to room temperature, adding 1400mL of ethanol to precipitate out polysaccharide, standing for 24h at 4 ℃, carrying out vacuum filtration, and carrying out vacuum freeze drying (drying for 72h and vacuum degree pressure range of 80 Pa) to obtain the dandelion flower polysaccharide extract with the solid content of 2.72g, wherein the content of the total polysaccharide can reach 62.01%, the contents of chlorogenic acid, caffeic acid and p-coumaric acid are 1.86%, 4.18% and 11.32% respectively, and the total flavone is about 9.51%. When the concentration of the dandelion flower polysaccharide extract is 1.0mg/mL, the bacteriostatic efficiency of staphylococcus aureus and escherichia coli is 63.63% and 22.56% respectively, and the proliferation efficiency of probiotics such as lactobacillus and bifidobacterium is 18.72% and 16.09% respectively.
Example 5
Weighing 133g of dandelion flower dry powder, adding 4000mL of distilled water into a glass extractor, extracting for 4h at a mechanical stirring speed of 100r/min under the boiling condition of 100 ℃, repeatedly extracting for 2 times, carrying out suction filtration while hot, collecting 2 times of filtrate, carrying out rotary evaporation and concentration at 55 ℃ until the concentrated solution is about 400mL, cooling the concentrated solution to room temperature, adding 1600mL of ethanol to precipitate polysaccharide, standing for 24h at 0 ℃, carrying out vacuum filtration, adding 900mL of distilled water to redissolve, and carrying out spray drying (the air inlet temperature is 135 ℃, the air outlet temperature is 80 ℃, the sample injection amount is 15mL/min, the drying time is 1h, and the range in a drying cavity is 0.09-0.1M Pa) to obtain the dandelion flower polysaccharide extract with the solid content of 2.23g, wherein the content of the total polysaccharide can reach 58.39%, the contents of chlorogenic acid, caffeic acid and p-coumaric acid are respectively 1.02%, 3.11% and 8.77%, and the total flavone content is about 7.87%. When the concentration of the dandelion flower polysaccharide extract is 1.0mg/mL, the bacteriostatic efficiency of staphylococcus aureus and escherichia coli is 60.01% and 19.48% respectively, and the proliferation efficiency of probiotics such as lactobacillus and bifidobacterium is 15.28% and 11.36% respectively.
The concentration of the dandelion flower polysaccharide extract expressed in the above embodiment means that it is heated with distilled water until it is completely dissolved. In addition, through GC-MS measurement, the dandelion flower is found to be composed of five monosaccharides, namely D-rhamnose, glucose, D-galactose, D-xylose and D-arabinose.
Comparative example
The dandelion flower contains higher phenols or flavonoids compounds than other tissue parts, and has stronger capability of inhibiting harmful bacteria. And monosaccharide compositions and structures in the polysaccharide are also different, and the produced polysaccharide also has different intestinal microecological regulation capacity. If the whole dandelion is adopted, a large amount of impurities with non-probiotic activity can be introduced by adopting the same extraction method (water extraction and alcohol precipitation method), and meanwhile, the obtained polysaccharide components and structures have differences, and the probiotic activity is not the same as that of the polysaccharide extract rich in polyphenol or flavone of pure petal tissues, and the specific effect is shown in figure 5.
The extraction method described in the comparative example is as follows:
weighing 120.0g of dried and pulverized dandelion flower or leaf or whole plant sample respectively, uniformly dispersing in 4000mL of distilled water, adjusting the temperature of the system to about 100 ℃, keeping the boiling state, and extracting for 4h under the condition of mechanical stirring (3000 r/min). Centrifuging the extractive solution at 8000g for 10min, and mixing the supernatants. After concentration by rotary evaporation, 4 volumes of absolute ethanol were added and left overnight at 4 ℃ to precipitate the polysaccharide. Centrifuging to collect the precipitated polysaccharide, and freeze-drying to obtain dandelion flower polysaccharide extract, dandelion leaf polysaccharide extract and dandelion whole plant extract. All the dandelion samples are called as Dayedandelion, and are obtained from the planting base of Sixue dandelion in Jiangsu Sixue in the last 3 th ten days of 2021 year at the same time, and are treated under the same processing conditions of drying (drying to constant weight at 50 ℃) and crushing (sieving by about 0.42mm through 60 meshes). The results of the measurement on the dandelion sample are shown in table 1, the monosaccharide composition in the dandelion flower polysaccharide extract is mainly mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and xylose, the molar ratio is 1; the monosaccharide composition in the whole dandelion polysaccharide extract mainly comprises mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and xylose, wherein the molar ratio of the monosaccharide to the galactose to the glucuronic acid to the xylose is 1.0. The dandelion leaf polysaccharide extract contains mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and xylose, and the molar ratio of the mannose to the rhamnose to the glucuronic acid to the galactose is 1.2. In addition, the total polysaccharide content in the dandelion flower extract is 66.8%, and the total polysaccharide content in the whole dandelion plant extract and the dandelion leaf extract is 61.3% and 59.86% respectively, which are lower than the total polysaccharide content in the dandelion flower extract.
TABLE 1 monosaccharide composition in polysaccharide extracts of different dandelion samples
Remarking: the total polysaccharide of the dandelion polysaccharide extract sample is determined by phenol-sulfuric acid method; the monosaccharide composition in the dandelion polysaccharide extract is determined by sequentially hydrolyzing with trifluoroacetic acid, derivatizing with PMP, and detecting with HPLC reverse phase chromatography under 254nm by combining with an external standard method.
In conclusion, the polysaccharide extract prepared from dandelion flowers serving as a raw material has strong antibacterial and probiotic activities, and can be directly applied to health-care products, medicines and cosmetics, so that the problem of safe use is solved. The invention adopts the freeze drying technology to prepare the concentrated solution of the polysaccharide active substance extracted from the dandelion flowers, furthest reserves the active ingredients in the polysaccharide extract of the dandelion flowers, reduces the use of organic solvents and the post-treatment requirement in the process of preparing the polysaccharide extract of the dandelion flowers, and reduces the cost, thereby being beneficial to the industrialized development of the dandelion. Therefore, the method for preparing the polysaccharide extract rich in the phenolic acid and the flavone by finely utilizing the dandelion flowers is a new way for processing and utilizing the dandelion, and has important significance for the basic research and development of the application of the dandelion.
Claims (3)
1. A taraxacum flower polysaccharide extract with antibacterial and probiotic activities is characterized in that taraxacum flower is used as a raw material, a water extraction and alcohol precipitation method is adopted to obtain the taraxacum flower polysaccharide extract, the taraxacum flower polysaccharide extract mainly contains polysaccharide, and is also rich in flavone such as luteolin, quercetin, luteolin and organic acid, and the organic acid comprises the following componentsp-coumaric acid, caffeic acid, chlorogenic acid; the dandelion flower polysaccharide extract with antibacterial and probiotic activities has the effect of treating staphylococcus aureus (I)S. aureus) Shigella bacteria (A), (B), (C)S. Castellani) Escherichia coli (E. Coli) ((E. Coli))E. Coli) And Salmonella bacteriaS. Enteritidis) Has inhibitory effect; for lactobacillus bacterium (b), (b)L. Bulgaricus) And Bifidobacterium (Bifidobacterium) One or both of them have a proliferative effect; the dandelion flower is fresh herba Taraxaci Mongolici produced by Jiangsu Siyang (herba Taraxaci)Taraxacum mongolicumHand. -mazz.), is used after being picked and sun-dried, and has the water content of less than 4 percent; crushing dandelion flowers, taking water as a solvent, stirring uniformly under a boiling condition, carrying out vacuum filtration while the dandelion flowers are hot, collecting filtrate, carrying out rotary evaporation, concentration and cooling to room temperature, adding absolute ethyl alcohol and a concentrated solution for alcohol precipitation, carrying out vacuum filtration, collecting alcohol precipitates, and carrying out vacuum freeze drying or spray drying after water redissolution to prepare a dandelion flower polysaccharide extract; the boiling condition refers to that water with the temperature of 100 ℃ is distilled water or deionized water, the solid-to-liquid ratio range of dandelion pollen to water is 1 to 60g/mL, the mechanical stirring speed is 10 to 1000r/min, and the stirring time is 1 to 240 min; the rotary evaporation concentration condition is that the temperature is 45-75 ℃, and the final concentration volume ranges from 200-300 mL; the volume ratio of the absolute ethyl alcohol to the concentrated solution is 1 to 10, and the alcohol precipitation is carried outThe temperature range was-4~4 ℃.
2. The method for preparing dandelion flower polysaccharide extract with both antibacterial and probiotic activities according to claim 1, wherein dandelion flowers are used as raw material, and the dandelion flower polysaccharide extract is obtained by water extraction and alcohol precipitation method; the dandelion flower is fresh herba Taraxaci Mongolici produced by Jiangsu Siyang (herba Taraxaci)Taraxacum mongolicumHand. -mazz.), is used after being picked and sun-dried, and has the water content of less than 4 percent; crushing dandelion flowers, taking water as a solvent, stirring uniformly under a boiling condition, carrying out vacuum filtration while the dandelion flowers are hot, collecting filtrate, carrying out rotary evaporation, concentration and cooling to room temperature, adding absolute ethyl alcohol and a concentrated solution for alcohol precipitation, carrying out vacuum filtration, collecting alcohol precipitates, and carrying out vacuum freeze drying or spray drying after water redissolution to prepare a dandelion flower polysaccharide extract; the boiling condition refers to that water with the temperature of 100 ℃ is distilled water or deionized water, the solid-to-liquid ratio of dandelion pollen to water is 1 to 60g/mL, the mechanical stirring speed is 10 to 1000r/min, and the stirring time is 1 to 240 min; the rotary evaporation concentration condition is that the temperature is 45 to 75 ℃, and the final concentration volume range is 200 to 300 mL; the volume ratio of the absolute ethyl alcohol to the concentrated solution is 1 to 10, and the temperature range of alcohol precipitation is-4~4 ℃.
3. The preparation method of the dandelion flower polysaccharide extract with antibacterial and probiotic activities according to claim 2, wherein the vacuum freeze-drying time range is 36 to 72h, the vacuum pressure range is 80 to 110Pa, or the spray-drying time range is 1 to 3h, and the vacuum pressure range is 0.09 to 0.1M Pa.
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| "中药蒲公英花提取物的抑菌性能研究";傅力明等;《山西医药杂志》;20150430;第44卷(第8期);第947-949页 * |
| "蒲公英不同器官多糖含量测定及抗氧化性研究";付学鹏等;《食品研究与开发》;20080331;第29卷(第3期);第26-28页 * |
| "蒲公英多糖的研究进展;刘炜熹等;《食品研究与开发》;20200531;第41卷(第10期);第214-219页 * |
| 付学鹏等."蒲公英不同器官多糖含量测定及抗氧化性研究".《食品研究与开发》.2008,第29卷(第3期),第26-28页. * |
| 傅力明等."中药蒲公英花提取物的抑菌性能研究".《山西医药杂志》.2015,第44卷(第8期),第947-949页. * |
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