CN105535033A - Method for extracting truffle polysaccharides from truffles - Google Patents
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- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The invention belongs to the technical field of traditional Chinese medicine and discloses a method for extracting truffle polysaccharides from truffles. The method comprises the following steps: ultrasonic extraction: black truffles are dried, ground and sieved, undersize powder is taken and subjected to ultrasonic extraction, water is adopted as an extraction solvent, the powder and the water are in a ratio being 1:40, the extraction temperature is 80 DEG C, the ultrasonic frequency is 100 kHz, extraction is performed for 2 h, filtering is performed after extraction, and filtering residues are extracted repeatedly; refining treatment: an extracted liquid is subjected to decompression concentration and left to stand until the room temperature is reached, absolute ethyl alcohol is added, the mixture is shaken up, left to stand and filtered, a precipitate is separated and washed with acetone, a washing liquid is removed, the precipitate is freeze-dried and ground, and white powder is obtained. The invention further discloses a compound prepared from the truffle polysaccharides and an application of the truffle polysaccharides in preparation of health care products with an immunity enhancement effect.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of method and application thereof of from Truffe, extracting Truffe polysaccharide.
Background technology
Immunity is the defense mechanism of human body self, be human bioequivalence and eliminate any foreign body (virus, antibacterial etc.) of external intrusion, process is old and feeble, damage, death, the ability of mutant cell and virus infected cell in the own cells of degeneration and identification and handling body.Immunology Today is thought, immunity is the physiological reaction of human bioequivalence and eliminating " dissident ".What perform this function in human body is immune system.Current truffle health-care product composition is single, does not have due effect.Up to now, people are finding a kind of Chinese medicine health care product making the raising immunity of simple, taking convenience.
Summary of the invention
Goal of the invention: in order to solve the problem, the object of the present invention is to provide a kind of method and application thereof of from Truffe, extracting Truffe polysaccharide.
Technical scheme: the object of the invention is by following scheme realize:
A kind of method extracting Truffe polysaccharide from Truffe, supersound extraction: Pollen pini thunbergii dew is dry, crush and screen, get minus sieve powder and carry out supersound extraction, water is Extraction solvent, solid-liquid ratio 1: 40, Extracting temperature is 80 DEG C, and supersonic frequency is 100kHz, extraction time 2h, filter after extracting, filtering residue carries out repeating to extract; Refinement treatment: by extracting solution concentrating under reduced pressure, puts to room temperature, adds anhydrous alcohol solution, shake up, and leaves standstill, and filter, sediment separate out, precipitate washing with acetone, discards cleaning mixture, precipitate lyophilization, pulverizes, obtains white powder.
The described method extracting Truffe polysaccharide from Truffe, Pollen pini thunbergii dew crushes and screens, and is sieving through 60 mesh sieves.
The described method extracting Truffe polysaccharide from Truffe, extracting solution concentrating under reduced pressure, pressure is 0.008 ~ 0.05Mpa, and temperature is 25 DEG C, and being concentrated into concentrated solution relative density is 1.25 ~ 1.36.
The described method extracting Truffe polysaccharide from Truffe, anhydrous alcohol solution consumption is 2 ~ 4 times of concentrated solution quality, leaves standstill 12 ~ 36h.
The described method extracting Truffe polysaccharide from Truffe, washing with acetone, acetone consumption is precipitate 0.5 ~ 3 times, washing with acetone 1 ~ 3 time.
A kind of have the truffle health-care product improving immunity, active component contained by it is prepared from by following raw materials in part by weight proportioning: Truffe polysaccharide 2 parts, Herba Equiseti Arvinsis 20 parts, the Radix Astragali 20 parts, Radix Codonopsis 18 parts, Radix Maesae Japonicae 14 parts, Herba Leibnitziae 20 parts, 6 parts, Radix Glycyrrhizae, and Truffe polysaccharide is that said method preparation obtains.
Described have the truffle health-care product improving immunity, preparation method step is: get above-mentioned other medical materials merging except Truffe polysaccharide, decoct with water twice, collecting decoction, when being concentrated into 65 DEG C, relative density is 1.10-1.20, add ethanol, stir, make alcohol content by volume percentage calculation reach 65-85%, leave standstill, filter, during filtrate reduced in volume to 65 DEG C, relative density is 1.20-1.30 and reclaims ethanol, and by concentrated solution spraying dry, gained granular powder is broken into dry extract, add Truffe polysaccharide and adjuvant, make required preparation.
Described have the truffle health-care product improving immunity, and in preparation method, decocting condition is: amount of water is the 8-12 times amount of medical material weight for the first time, decocts 1-2h, and second time amount of water is the 6-10 times amount of medical material weight, decocts 1-2h.
Described have the truffle health-care product improving immunity, and in preparation method, spray drying condition is: inlet temperature is 90-110 DEG C, and leaving air temp is 60-90 DEG C, and temperature of charge is 60-90 DEG C, and atomizing pressure is 0.2-0.5 MPa, and spray velocity is 1-10ml/s.
Described have the application of truffle health-care product in preparation raising immunity health product improving immunity.
Above-mentioned Truffe is Pollen pini thunbergii dew, (Latin is called Tubermelanosporum to Pollen pini thunbergii dew, English is called perigordtruffle), also claim truffles, be a kind of wild edible fungi being grown on underground, appearance is rugged and rough, color and luster is between dark-brown and black, in small embossment shape, throughout the texture of Lycoperdon polymorphum Vitt or light/dark balance and white, its abnormal smells from the patient is special.
Herba Equiseti Arvinsis, Latin is called Equisetumarvense), herbaceos perennial, Equisetales, Equisetaceae, Equisetum, nature and flavor: bitter, cool.Heat clearing away, removing heat from blood, cough-relieving, diuresis.Control epistaxis, spit blood, spitting of blood, has blood in stool, metrorrhagia, traumatic hemorrhage, cough and asthma, gonorrhea, conjunctivitis.Aerial parts (Herba Equiseti Arvinsis) is bitter, puckery, cool.Clearing away heat and promoting diuresis, hemostasis, suppressing the hyperactive liver improving eyesight, relieving cough and asthma.For nose nephew, enterorrhagia, spitting of blood, hemorrhoid hemorrhage, menorrhagia, stranguria, fracture, cough with asthma, conjunctival congestion and swelling pain.The present invention is with all herbal medicine.
Radix Maesae Japonicae is Myrsinacea Maesa plant Radix Maesae Japonicae Maesajaponica (Thunb.) Moritzi, is used as medicine with root, leaf.The whole year can adopt, and cleans and dries.To dispel the wind diuresis, hemostasis, detumescence.Root: headache, lumbago, edema, ascites; Leaf: wound hemorrhage is controlled in external.The present invention is used as medicine with root.
Herba Leibnitziae is the herb of feverfew Herba Leibnitziae, perennial herb.Be born in the dark and damp places such as roadside, hillside, Lin Bian, meadow, limes marginis, be distributed in the ground such as Chinese north and south.There is clearing away heat-damp and promoting diuresis; Effect of removing toxic substances and promoting subsidence of swelling.Cure mainly cough due to lung-heat; Damp-heat dysentery; Pyretic stranguria; Rheumatic arthritis; Carbuncle furuncle and phyma poison; Ecthyma; Early venom; Burn and scald; Traumatic hemorrhage.All the other are pharmacopeia kind.
Beneficial effect: immunity is low is exactly positive QI-insufficiency by traditional Chinese medical science saying.Tonify deficiency righting is not only wanted in treatment, also answers blood circulation promoting and blood stasis dispelling, heat-clearing and toxic substances removing.Therefore in we, Herba Equiseti Arvinsis, Herba Leibnitziae heat-clearing and toxic substances removing are monarch drug, and Radix Maesae Japonicae cooling blood for hemostasis is ministerial drug, Truffe, the Radix Astragali, Radix Codonopsis, Radix Glycyrrhizae QI invigorating, clearing away heat-damp and promoting diuresis, and dispel the wind as adjuvant, all medicines are harmonious, and play benefiting QI for activating blood circulation altogether, for improving immunity.
Detailed description of the invention
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1: Pollen pini thunbergii dew powder crosses 60 mesh sieves, gets minus sieve powder 100g, adds water 1500g, at 20 DEG C, KQ-100DV type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.), supersonic frequency 30kHz, extraction time 0.5h.Extraction time 3 times, filters, and is merged by three extracting solution, at vacuum pressure 0.08MPa, at temperature 25 DEG C, be evaporated to relative density 1.36, put to room temperature, add 3 times of anhydrous alcohol solution, shake up, leave standstill 12h, filter, sediment separate out, with 0.5 times of washing with acetone 3 times, discard cleaning mixture, precipitate is lyophilization at-50 DEG C, pulverizing according to a conventional method, obtains 19.96g white powder.Polysaccharide yield is 8.79% after measured.
Embodiment 2: Pollen pini thunbergii dew powder crosses 60 mesh sieves, and get minus sieve powder 100g, add water 3000g, Extracting temperature is 50 DEG C, supersonic frequency 80kHz, extraction time 2h.Extraction time 2 times, filters, is merged by extracting solution, be evaporated to relative density 1.18, put to room temperature, add 4 times of anhydrous alcohol solution, shake up, leave standstill 24h, filter, sediment separate out, with 1.5 times of washing with acetones 2 times, discard cleaning mixture, precipitate is lyophilization at-50 DEG C, pulverizing according to a conventional method, obtains 20.32g white powder.Polysaccharide yield is 8.97% after measured.
Embodiment 3: Pollen pini thunbergii dew powder crosses 60 mesh sieves, and get minus sieve powder 100g, add water 4000g, Extracting temperature is 80 DEG C, supersonic frequency 100kHz, 2 hours extraction times.Extraction time 1 time, filters, extracting solution is evaporated to relative density 1.29, put to room temperature, add 3 times of anhydrous alcohol solution, shake up, leave standstill 36 hours, filter, sediment separate out, with 3 times of washing with acetones 1 time, discards cleaning mixture, precipitate is lyophilization at-50 DEG C, pulverizing according to a conventional method, obtains 22.15g white powder.Polysaccharide yield is 9.36% after measured.
Embodiment 4: get Herba Equiseti Arvinsis 20g, Radix Astragali 20g, Radix Codonopsis 18g, Radix Maesae Japonicae 14g, Herba Leibnitziae 20g, Radix Glycyrrhizae 6g medical material mixes, decoct with water twice, first time adds water as 8 times amount of medical material weight, decoct 1.5h, second time adds water as 6 times amount of medical material weight, decoct 1h, collecting decoction, filter, filtrate is concentrated into relative density 1.10 (65 DEG C), adding ethanol in proper amount makes alcohol content (volume fraction) reach 65%, stir, leave standstill 24 hours, filter, filtrate recycling ethanol is also concentrated into the extractum of relative density 1.10 (65 DEG C), it is spray-dried that (condition is inlet temperature is 100 DEG C, leaving air temp is 80 DEG C, temperature of charge is 70 DEG C, atomizing pressure is 0.2 MPa, spray velocity is 5ml/s.), be ground into dry extract, then add Truffe polysaccharide 2g, dextrin 50g, with appropriate 80% ethanol wet, soft material processed, cross 14 mesh sieves and granulate, 70 DEG C of dryings, 60 order granulate, obtain granule.
Embodiment 5: get Herba Equiseti Arvinsis 20g, Radix Astragali 20g, Radix Codonopsis 18g, Radix Maesae Japonicae 14g, Herba Leibnitziae 20g, Radix Glycyrrhizae 6g medical material mixes, decoct with water twice, add water as 10 times amount of medical material weight time, decoct 2h, second time adds water as 8 times amount of medical material weight, decoct 1h, collecting decoction, filter, filtrate is concentrated into relative density 1.15 (65 DEG C), adding ethanol in proper amount makes alcohol content (volume fraction) reach 75%, stir, leave standstill 24 hours, filter, filtrate recycling ethanol is also concentrated into the extractum of relative density 1.25 (65 DEG C), it is spray-dried that (condition is inlet temperature is 120 DEG C, leaving air temp is 90 DEG C, temperature of charge is 80 DEG C, atomizing pressure is 0.3 MPa, spray velocity is 7.5/s.), be ground into dry extract, then add Truffe polysaccharide 2g, starch 50g, mix homogeneously, with appropriate 80% ethanol wet, soft material processed, crosses 30 mesh sieves and granulates, in 70 ~ 80 DEG C of dryings, with 60 mesh sieve granulate, and tabletting, bag fat clothing, subpackage, outer package, censorship is qualified, obtains finished product.
Embodiment 6: get Herba Equiseti Arvinsis 20g, Radix Astragali 20g, Radix Codonopsis 18g, Radix Maesae Japonicae 14g, Herba Leibnitziae 20g, Radix Glycyrrhizae 6g medical material mixes, decoct with water twice, first time adds water as 10 times amount of medical material weight, decoct 1.5h, second time adds water as 8 times amount of medical material weight, decoct 1.5h, collecting decoction, filter, filtrate is concentrated into relative density 1.15 (65 DEG C), adding ethanol in proper amount makes alcohol content (volume fraction) reach 75%, stir, leave standstill 24 hours, filter, filtrate recycling ethanol is also concentrated into the extractum of relative density 1.25 (65 DEG C), it is spray-dried that (condition is inlet temperature is 120 DEG C, leaving air temp is 90 DEG C, temperature of charge is 90 DEG C, atomizing pressure is 0.4 MPa, spray velocity is 7.5/s.), be ground into dry extract, cross No. 6 sieves, add Truffe polysaccharide 2g, reinstall No. 1 capsule finished product.
Embodiment 7: the present invention improves the pharmacodynamic study of immunity
Experiment purpose: with the gavage mice 30d of the present invention of variable concentrations, compares the present invention and normal group mice by the difference of the splenic lymphocyte proliferation of ConA induction, judges whether the present invention strengthens splenic lymphocyte proliferation.
Laboratory animal: Kunming mouse, male, body weight 18-22g, Shanghai Slac Experimental Animal Co., Ltd. provides, production licence: SCXK (Shanghai) 2007-0005.
Experimental agents: prepare by the preparation method of above-mentioned granule embodiment 4.
Experimental procedure: mice, by the grouping of body weight stratified random, often organizes 12.As shown in the table, normal group gavage feeds water; Levamisole hydrochloride group gavage levamisole hydrochloride; The other gavage 1,2,4gkg of the basic, normal, high dosage component of compound recipe
-1inventive samples, the continuous gastric infusion 30d of each group.Asepticly get spleen, conveniently make individual cells suspension, adjustment cell concentration is 5 × 10
5individual/mL.Be added in by splenocyte suspension in 24 orifice plates, every hole 1mL, every animal establishes two multiple holes, and a hole is normally cultivated (adding 50uLPRMI1640 culture fluid); One hole adds 50uL concanavalin A, Con A (ConA) liquid (final concentration is 4.76ug/mL).Be placed in 5%CO
2, 37 DEG C of CO
248h is cultivated in incubator.Experiment terminates front 4h, and every hole sucks supernatant 700uL, then adds 700uL not containing the PRMI1640 culture fluid of hyclone, and every hole adds the MTT liquid of 50uL5mg/mL simultaneously.After cultivation terminates, every hole adds 1mL acid isopropyl alcohol, and the even crystallization to purple of careful piping and druming is all dissolved.Be dispensed in 96 well culture plates before detection, 4 multiple holes are done in every hole, and at 570nm wavelength, place measures its optical density value.
Experimental result: of the present invention group of mouse spleen lymphocyte is significantly higher than normal group mice by the rate of increase conversion values after ConA induction, as shown in the table.
Table 1 the present invention is on the impact of spleen lymphocyte proliferation
n=12)
Note: compared with normal group
★ ★p < 0.01,
★p < 0.05.
Conclusion: the present invention can strengthen spleen lymphocytic hyperplasia, therefore the present invention can improve immunity, can be used for preparation and improves efficacy of immunity health product.
Claims (10)
1. one kind is extracted the method for Truffe polysaccharide from Truffe, it is characterized in that: supersound extraction: Pollen pini thunbergii dew is dry, crush and screen, get minus sieve powder and carry out supersound extraction, water is Extraction solvent, solid-liquid ratio 1: 40, Extracting temperature is 80 DEG C, and supersonic frequency is 100kHz, extraction time 2h, filter after extracting, filtering residue carries out repeating to extract; Refinement treatment: by extracting solution concentrating under reduced pressure, puts to room temperature, adds anhydrous alcohol solution, shake up, and leaves standstill, and filter, sediment separate out, precipitate washing with acetone, discards cleaning mixture, precipitate lyophilization, pulverizes, obtains white powder.
2. from Truffe, extract the method for Truffe polysaccharide according to claim 1, it is characterized in that: Pollen pini thunbergii dew crushes and screens, and is sieving through 60 mesh sieves.
3. from Truffe, extract the method for Truffe polysaccharide according to claim 1, it is characterized in that: extracting solution concentrating under reduced pressure, pressure is 0.008 ~ 0.05Mpa, and temperature is 25 DEG C, and being concentrated into concentrated solution relative density is 1.25 ~ 1.36.
4. from Truffe, extract the method for Truffe polysaccharide according to claim 1, it is characterized in that: anhydrous alcohol solution consumption is 2 ~ 4 times of concentrated solution quality, leave standstill 12 ~ 36h.
5. from Truffe, extract the method for Truffe polysaccharide according to claim 1, it is characterized in that: washing with acetone, acetone consumption is precipitate 0.5 ~ 3 times, washing with acetone 1 ~ 3 time.
6. one kind has the truffle health-care product improving immunity, it is characterized in that, active component contained by it is prepared from by following raw materials in part by weight proportioning: Truffe polysaccharide 2 parts, Herba Equiseti Arvinsis 20 parts, the Radix Astragali 20 parts, Radix Codonopsis 18 parts, Radix Maesae Japonicae 14 parts, Herba Leibnitziae 20 parts, 6 parts, Radix Glycyrrhizae, and Truffe polysaccharide is that the method preparation of the claims 1 obtains.
7. there is truffle health-care product as claimed in claim 6 that improve immunity, it is characterized in that, preparation method step is: get above-mentioned other medical materials merging except Truffe polysaccharide, decoct with water twice, collecting decoction, when being concentrated into 65 DEG C, relative density is 1.10-1.20, add ethanol, stir, make alcohol content by volume percentage calculation reach 65-85%, leave standstill, filter, during filtrate reduced in volume to 65 DEG C, relative density is 1.20-1.30 and reclaims ethanol, by concentrated solution spraying dry, gained granular powder is broken into dry extract, add Truffe polysaccharide and adjuvant, make required preparation.
8. there is truffle health-care product as claimed in claim 7 that improve immunity, it is characterized in that, in preparation method, decocting condition is: amount of water is the 8-12 times amount of medical material weight for the first time, decocts 1-2h, second time amount of water is the 6-10 times amount of medical material weight, decocts 1-2h.
9. have the truffle health-care product improving immunity as claimed in claim 7, it is characterized in that, in preparation method, spray drying condition is: inlet temperature is 90-110 DEG C, leaving air temp is 60-90 DEG C, temperature of charge is 60-90 DEG C, and atomizing pressure is 0.2-0.5 MPa, and spray velocity is 1-10ml/s.
10. there is truffle health-care product as claimed in claim 6 that improve immunity and improving the application in immunity health product.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105535032A (en) * | 2016-03-04 | 2016-05-04 | 陈爱华 | Extraction method of truffle polysaccharides and health-care truffle product |
| CN107536019A (en) * | 2017-10-23 | 2018-01-05 | 深圳深深爱健康科技有限公司 | A kind of preparation method of black pine nano powder |
| CN112535287A (en) * | 2020-12-07 | 2021-03-23 | 皖西学院 | Compound edible fungus oral liquid and preparation process thereof |
| CN112690311A (en) * | 2020-12-28 | 2021-04-23 | 广州市锦华食品有限公司 | Black truffle polysaccharide slurry-popping cookie biscuits and making method thereof |
| CN115368481A (en) * | 2022-08-08 | 2022-11-22 | 北京工商大学 | Preparation method and application of multifunctional truffle polysaccharide |
Citations (1)
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| CN101580551B (en) * | 2009-06-23 | 2011-11-30 | 宽甸玉竹农副产品有限公司 | Method for extracting polygonatum odoratum polysaccharide |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101580551B (en) * | 2009-06-23 | 2011-11-30 | 宽甸玉竹农副产品有限公司 | Method for extracting polygonatum odoratum polysaccharide |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105535032A (en) * | 2016-03-04 | 2016-05-04 | 陈爱华 | Extraction method of truffle polysaccharides and health-care truffle product |
| CN107536019A (en) * | 2017-10-23 | 2018-01-05 | 深圳深深爱健康科技有限公司 | A kind of preparation method of black pine nano powder |
| CN112535287A (en) * | 2020-12-07 | 2021-03-23 | 皖西学院 | Compound edible fungus oral liquid and preparation process thereof |
| CN112690311A (en) * | 2020-12-28 | 2021-04-23 | 广州市锦华食品有限公司 | Black truffle polysaccharide slurry-popping cookie biscuits and making method thereof |
| CN115368481A (en) * | 2022-08-08 | 2022-11-22 | 北京工商大学 | Preparation method and application of multifunctional truffle polysaccharide |
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