CN1969682A - Application of bamboo shoot polysaccharide in preparation of health food - Google Patents
Application of bamboo shoot polysaccharide in preparation of health food Download PDFInfo
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- CN1969682A CN1969682A CNA2006101544770A CN200610154477A CN1969682A CN 1969682 A CN1969682 A CN 1969682A CN A2006101544770 A CNA2006101544770 A CN A2006101544770A CN 200610154477 A CN200610154477 A CN 200610154477A CN 1969682 A CN1969682 A CN 1969682A
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Abstract
The invention relates to the novel use of bamboo shoot crude polysaccharides in preparing health food having immunological regulation function, wherein bamboo shoots or dried bamboo shoots are used as raw materials to extract crude polysaccharides therein, the active constituents and acceptable auxiliary materials are employed to develop health food preparations including capsules, drop pills, tablets, granules, powders and oral liquids.
Description
Technical field:
The present invention relates to the purposes of bamboo shoot polysaccharide, especially the purposes in health food.
Technical background:
The bamboo shoots polysaccharide is the soluble polysaccharide that extracts from discarded bamboo shoot clothing, bamboo shoot head.The polymer that polysaccharide is made up of 10 above monose, as the dietary fiber of solubility, it has possessed the general physiological function of dietary fiber, and modern science proved already, dietary fiber has functions such as the hypertension of control, coronary heart disease etc. and removing external source harmful substance.All the shortage with dietary fiber is relevant for the sick morbidity as hypertension, high fat of blood, obesity etc. of many modern civilizations.Simultaneously, polysaccharide has prior physiologically active, the identification of the adjusting of polysaccharide and immunologic function, cell and cell toward contact.The transportation of intercellular substance, the Clinics and Practices of cancer etc. have confidential relation.Polysaccharide pharmaceutically still is being a kind of good adjuvant.It can also control the division and the differentiation of cell in addition, diversified biologically actives such as the growth of adjusting cell and aging.At present, in the ascendant to the research of the chemical research of polysaccharide and biological function, the potential function of separate sources polysaccharide is determined and need further be studied.
At present the physiological function of the functional polysaccharide of finding is mainly aspect following two:
One, antitumor
People have obtained multiple polysaccharide with antitumor activity from different biomaterials, has strong antitumor activity as the lentinan that from mushroom, obtains, it suppresses cancer and antitumous effect is not to directly act on the transplantability cancer cell, but has an effect by host's adjusting.
Two, strengthen immunization
Confirmed that different polysaccharide has different immunologic enhancements.Lentinan is desirable immunopotentiating agent, and it stimulates the participation organism immune response as the adjuvant and the helper cell of T celluar localization.Have now found that it has the high degree of specificity immunological enhancement.Tremella polysaccharides can promote lymphocytic conversion, strengthens the phagocytic function of Turnover of Mouse Peritoneal Macrophages, obviously promotes the synthetic of hepatic protein nucleic acid and promotes hemopoietic function of bone marrow, improves humoral immunity.Blackfungus polyhexose can improve the phagocytic index and the percentage of mouse macrophage effectively.Pharmacological evaluation proves that tea polysaccharide has the enhancing immunologic function, promotes the effect of mononuclear phagocyte system phagocytic function.Studies show that: tea polysaccharide is a kind of immunopotentiating agent, can obviously promote the phagocytic activity of macrophage, the immune organ spleen weight is increased, the polysaccharide of other polysaccharide such as extractions such as garlic, radish has different humidifications respectively to T lymphocyte, bone-marrow-derived lymphocyte and macrophage, can strengthen the body cell immunologic function.
Though the STUDY ON POLYSACHAROSE document is more, the bamboo shoots STUDY ON POLYSACHAROSE is comparatively rare, after the literature search, and rarely seen discrete functions report, the existing function of report is antitumor (zoopery).
Summary of the invention
The object of the present invention is to provide the new purposes of bamboo shoot polysaccharide, i.e. purposes in the health food functional localization.
Characteristics of the present invention are the application of bamboo shoot polysaccharide in health food.
The concrete bamboo shoot polysaccharide of the present invention has new application in the health food of immunoloregulation function in preparation.
The present invention is a raw material with bamboo shoots or its dried product dried bamboo shoots, extract thick polysaccharide wherein, it is developed to the suitable health food preparation with immunoloregulation function as acceptable other raw materials and auxiliary material on active component and the food technology, as capsule (soft capsule, hard shell capsules), pill, tablet, granule, powder, oral liquid etc.
In order to understand essence of the present invention better, illustrate that with the result of research of the immunoloregulation function of bamboo shoot polysaccharide and acute toxicity testing it has purposes in the health food of immunoloregulation function in preparation below.
One: the immunological regulation test of bamboo shoot polysaccharide: the mouse spleen lymphocyte conversion test (mtt assay) that ConA induces
I: test principle: one. principle: after mitogen stimulates blast transformation takes place when T lymphocyte receptor ConA causes, living cells particularly proliferative cell develops the color (formazan) crystallization of praising by the first moon that MTT (a kind of flaxen azoles salt) is decomposed into blue purple by the mitochondria hydrolase, and its OD value can reflect the propagation situation of cell.
II: test equipment and reagent:
| Desinfection chamber, superclean bench | N H 4CI | Bromogeramine |
| Positive pressure devices | RPMI1640 | Calf serum |
| Filter (0.22 μ m, 0.45 μ m) | Glutamine | Penicillin, streptomysin |
| Heavy sheet glass bottle (500ml, 250ml, 100ml, 50ml) penicillin Bottle and closure | N aHCO 3 | Hanks liquid |
| Various droppers, water dropper, suction pipe, suction nozzle, glass-tube, pipette | ConA | Platform is expected blue dyeing liquor |
| 200 eye mesh screens | N aCl | Bamboo shoot polysaccharide (freeze drying, vacuum drying) |
| The eye scissors of tweezers, scissors, straight peen and elbow, ophthalmology tweezer number | KCl | |
| Culture dish several | MTT | |
| Micro sample adding appliance (1ml, 20ul, 200ul) | Isopropyl alcohol | |
| 96 porocyte plates | Ethanol | |
| 24 porocyte plates | NaCl | |
| HCl | ||
| Above equipment and reagent all need be divided into ultraviolet sterilization, moist heat sterilization, filtration sterilization etc. through sterilization according to circumstances when experiment. | ||
III: test procedure:
1. reagent preparation:
1.1 preparation RPMI1640 complete culture solution: the explanation according to RPMI1640 prepares the RPMI1640 nutrient solution, filtration sterilization is with preceding adding 10% calf serum, 1% glutamine (200mmol/L), penicillin (100U/ml), streptomysin (100ug/L) and 5 * 10
-5The mol/L 2 mercapto ethanol is transferred pH7.0-7.2, i.e. complete culture solution with the HCl of aseptic 1mol/L or the NaOH of 1mol/L.
1.2ConA liquid is mixed with the solution of 100ug/mL with distilled water, filtration sterilization is preserved at low temperature refrigerator (20 ℃).With preceding being mixed with 5 μ l/ml
1.3PBS liquid: 0.02M pH7.2 1L
1.4 0.155M NH
4Cl 0.83g constant volume is to 100ml
1.5 aseptic Hanks liquid is transferred pH7.2-7.4 with preceding aseptic NaHCO3 with 3.5%.
1.6 MTT liquid 5mgMTT is dissolved among the PBS of 1mLpH7.2, now joins use.
1.7 add the HCL of 4ml 1mol/L in the acid isopropyl alcoholic solution 96mL isopropyl alcohol, face and use preceding preparation.
1.8 bamboo shoot polysaccharide stoste (100mg/ml) preparation
In bamboo shoot polysaccharide powder 40g → PBS400ml → ice cube, stirring and evenly mixing 30min → ultrasonic processing (ice bath) → 4 ℃ 15000rpm 10min is centrifugal → get supernatant, and 10 layers of filtered through gauze → 0.45 μ m filter membrane suction filtration → aseptic 0.22 μ m filter membrane suction filtration → 4 ℃ preservation → with preceding 33.33mg/ml, the 11.11mg/ml of being mixed with respectively
2. splenocyte suspension preparation:
Put to death mouse, 75%Alc soaks, and cuts off, and the aseptic spleen of getting places the little plate that fills an amount of aseptic Hanks liquid, gently spleen is torn up with tweezers, makes the individual cells suspension.Filter through 200 eye mesh screens, wash with Hanks liquid, centrifugal 10min (1500r/min) adds 1mlNH
4Cl3.5min adds Hanks liquid, and centrifugal 10min (1500r/min) uses the centrifugal 10min of Hanks liquid (1500r/min) to wash once again.Then cell is suspended in the complete culture solution of 2mL, with the blue dyeing counting viable count of platform phenol (should more than 95%), adjusting cell concentration be 2 * 10
6Individual/Ml.
3. lymphproliferation response:
Cell suspension branch hole is added in 24 well culture plates every hole 1Ml.Add corresponding reagent again.
| Blank PBS50 μ l | High concentration freeze drying bamboo shoot polysaccharide powder 50 μ l | High concentration vacuum drying bamboo shoot polysaccharide powder 50 μ l |
| ConA contrasts 50 μ l | Low concentration freeze drying bamboo shoot polysaccharide powder 50 μ l | High concentration vacuum drying bamboo shoot polysaccharide powder 50 μ l |
| The collaborative contrast of ConA PBS50 μ l+ConA25 μ l | High concentration freeze drying bamboo shoot polysaccharide powder 50 μ l+ConA25 μ l | High concentration vacuum drying bamboo shoot polysaccharide powder 50 μ l+ConA25 μ l |
Put 5%CO2, cultivate 72h for 37 ℃.Cultivate and finish preceding 4h, supernatant 0.7mL is inhaled in every hole gently, adds the RPMI1640 nutrient solution that 0.7mL does not contain calf serum, adds MTT (5mg/mL) 50ul/ hole simultaneously, continues to cultivate 4h.After cultivating end, every hole adds 1mL acid isopropyl alcohol, and the piping and druming mixing dissolves purple crystal fully.Divide then to install in 96 well culture plates, enzyme-linked immunosorbent assay instrument is used as parallel sample in each hole packing 3 hole.Measure OD value with the 570nm wavelength.
IV: experimental result and judgement:
1. experimental result:
2. judge:
From experimental result as can be seen: the OD of vacuum drying bamboo shoot polysaccharide powder test solution
570Value is apparently higher than other value.From this experimental result, vacuum drying bamboo shoot polysaccharide powder test solution has the function that promotes cellular immunity.
Two: the acute toxicity test of bamboo shoot polysaccharide:
1. experiment material and method
1.1 animal used as test Kunming kind small white mouse 18～22g, male and female half and half, the quarantine back is standby.
1.2 the experimental technique mouse is divided into 6 dosage groups by the body weight equilibrium, 20 every group.Fasting be can't help water 12 hours before the experiment, according to the preliminary experiment result, be divided into six dosage group: 11.03g/Kg, 13.00g/Kg, 14.25g/Kg, 16.00g/Kg, 18.40g/Kg, 20.75g/Kg, adjacent doses ratio is 0.88, dilution is to mouse stomach, to observing 4h, 24h, 48h, 72h, each observation in 7 days behind the sample continuously once.Record small white mouse toxic reaction and death toll after 7 days.
2. result of the test:
| Dosage (g/Kg) | Number of animals (only) | Dead animal number (only) (♂: ♀) | The death rate (%) |
| 11.03 | 20 | 0(0∶0) | 0 |
| 13.00 | 20 | 0(0∶0) | 0 (but poisoning symptom is arranged) |
| 14.25 | 20 | 10(2∶3) | 50 |
| 16.00 | 20 | 12(3∶3) | 60 |
| 18.40 | 20 | 16(3∶5) | 80 |
| 20.75 | 20 | 20(5∶5) | 100 |
After observing in continuous seven days, not dead again.Obvious pathology is not seen in the dead animal perusal that becomes celestial.Experiment draws this sample LD50=14.25g/Kg (90% fiducial limit) thus.Press acute toxicity grading criteria and judge the true border of this sample non-toxic type.
In sum, bamboo shoot polysaccharide has good result in the function aspects of external promotion cellular immunity, and toxic and side effect is little, and the excellent development application prospect is arranged.
The used raw material of bamboo shoot polysaccharide of the present invention is the old root or the bamboo shoot clothing of bamboo shoots and dried product thereof, but not the tender point of dried bamboo shoots of higher commodity value is arranged, and this will improve the effective rate of utilization of bamboo shoots to a certain extent.Method technology of the present invention is simple, meets the requirement of Chinese Pharmacopoeia, need not special installation.The method that the common water of bamboo shoot polysaccharide employing among the present invention is carried, film concentrates or the low pressure film concentrates, alcohol precipitation extraction method etc. is suitable for suitability for industrialized production is prepared.The bamboo shoot polysaccharide that obtains is light lark powder.
Above-mentioned bamboo shoot polysaccharide extraction method the steps include: to calculate by weight, take by weighing 0.1～20 part on drying bamboo shoots, add 0.5～200 part in water, heating and refluxing extraction 3～5 hours, temperature are controlled at 60～105 ℃ (heating temperature in early stage can be provided with highlyer, and the later stage is as long as keep temperature), the extract vacuum is concentrated (vacuum-0.4--0.01Mpa, temperature 55-70 ℃), being concentrated to concentration is 25-55 degree (with refractometer), cooling.Under agitation, slowly add concentrate in the 3-15 times of volume of ethanol or 3-15 times of volume of ethanol slowly added in the concentrate of stirring (speed of agitator is 30-400 rev/min), precipitate 0.2-10 hour, sediment is carried out vacuum drying or freeze drying, obtain bamboo shoot polysaccharide.
Bamboo shoot polysaccharide has significantly immunoloregulation function, and tasteless odorless, can be easily and other raw-food material compatibilities, or separately as the major ingredient development, make bamboo shoots ground active ingredient be fully utilized, thereby filled up the blank that China bamboo shoots utilize, had practical value.
The specific embodiment
Following example is in order to further specifying the present invention, but do not limit the scope of the invention thus.
Embodiment 1:
Calculate by weight, take by weighing drying bamboo shoots 500 grams, put into the refluxing extraction jar, add 2500 ml waters, the heater heating, and in reflux condenser, connect condensed water, after the feed liquid boiling, turn down heater, make that feed liquid is little boils, extracted 4 hours, obtain the dark-brown extract after the cooling; Extract is put into the vacuum film evaporimeter, and control vacuum is-0.08, and temperature is 65 ℃, and feed liquid is evaporated to 40 degree (with refractometer), obtains about 200 milliliters of concentrates; 1000 milliliter 95% edible ethanol is put into 2000 ml beakers, insert agitator, open and stir, adjust rotating speed at 100 rev/mins, concentrate joined with the speed of 20 ml/min in 95% the edible ethanol, add the material back and continue to stir more than 30 minutes, polysaccharide is fully precipitated; Stop stirring, precipitate 4 hours, the sucking-off supernatant liquor will precipitate with filter paper and leach, and will precipitate with 50 milliliters of absolute ethanol washings twice, and (55 ℃ of temperature, vacuum-0.08Mpa) obtain 5 canescence bamboo shoot polysaccharide powder about restraining in vacuum drying.
Embodiment 2:
Calculate by weight, take by weighing 50 kilograms on drying bamboo shoots, put into multi-function extractor, adding 300 kilograms purifies waste water, open the heating of jacket steam valve, and in reflux condenser, connect condensed water, after the feed liquid boiling, turn down steam valve, material is remained on more than 70 ℃, extracted 6 hours, slag is purified waste water same extraction once with 200 kilograms again, merge extract, obtain the dark-brown extract after the cooling; Extract is put into vacuum concentration pot, and control vacuum is-0.08, and temperature is 70 ℃, and feed liquid is evaporated to 38 degree (with refractometer), obtains about 60 liters of concentrates; 400 liter 95% edible ethanol is put into 500 liters of recycling cans, open to stir, adjust rotating speed, concentrate is joined with 1 liter/minute speed in 95% the edible ethanol, add the material back and continue to stir 60 minutes, polysaccharide is fully precipitated at 50 rev/mins; Stop stirring, precipitate 4 hours, the sucking-off supernatant liquor is squeezed into tripod pendulum type batch centrifugal with lower floor's feed liquid, is filter medium with the industrial filter cloth, precipitation is leached, collecting precipitation will precipitate with 10 liters of absolute ethanol washings twice, will precipitate vacuum drying (45 ℃ of temperature after centrifugal, vacuum-0.09Mpa), obtain the canescence bamboo shoot polysaccharide powder about 650 grams.
Claims (1)
1, bamboo shoot polysaccharide has application in the health food of immunoloregulation function in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006101544770A CN1969682A (en) | 2006-11-02 | 2006-11-02 | Application of bamboo shoot polysaccharide in preparation of health food |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006101544770A CN1969682A (en) | 2006-11-02 | 2006-11-02 | Application of bamboo shoot polysaccharide in preparation of health food |
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| CN1969682A true CN1969682A (en) | 2007-05-30 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107708445A (en) * | 2015-06-15 | 2018-02-16 | 玉井乃 | The manufacture method of bamboo extractive from fermentative and immunity effect agent |
| CN107827994A (en) * | 2017-12-05 | 2018-03-23 | 无锡盛雅生物科技有限公司佛山分公司 | A kind of extracting method of winter bamboo shoot polysaccharide |
-
2006
- 2006-11-02 CN CNA2006101544770A patent/CN1969682A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107708445A (en) * | 2015-06-15 | 2018-02-16 | 玉井乃 | The manufacture method of bamboo extractive from fermentative and immunity effect agent |
| US10820612B2 (en) | 2015-06-15 | 2020-11-03 | Osamu Tamai | Method for producing fermented bamboo extract and immunostimulating agent |
| CN107827994A (en) * | 2017-12-05 | 2018-03-23 | 无锡盛雅生物科技有限公司佛山分公司 | A kind of extracting method of winter bamboo shoot polysaccharide |
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Open date: 20070530 |