CN107586346A - One kind tree butterfly polysaccharide and preparation method and application - Google Patents
One kind tree butterfly polysaccharide and preparation method and application Download PDFInfo
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- CN107586346A CN107586346A CN201710778774.0A CN201710778774A CN107586346A CN 107586346 A CN107586346 A CN 107586346A CN 201710778774 A CN201710778774 A CN 201710778774A CN 107586346 A CN107586346 A CN 107586346A
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Abstract
The invention belongs to the technical field of food, discloses a kind of tree butterfly polysaccharide and preparation method and application.Methods described is：Butterfly grinding and sieving will be set, adds distilled water, will be ultrasonically treated, heating and refluxing extraction, will be filtered, filtrate concentration, obtains Thick many candies solution；The macroporous absorbent resin by pretreatment, decolorization, filtering are added, and washes out the polysaccharide adsorbed in resin repeatedly with water, polysaccharide solution after decolouring is merged, is concentrated under reduced pressure, precipitating reagent is added and is precipitated, centrifugation, sediment and drying are collected, obtain setting butterfly Thick many candies.The polysaccharide of the present invention has anti-oxidant, antitumor and immune-enhancing activity, and dosage is low, and the toxic side effect under same dosage is relatively low.
Description
Technical field
The invention belongs to the technical field of food, and in particular to one kind tree butterfly polysaccharide and preparation method and application.
Background technology
Polysaccharide is a kind of important biological polymeric compound in addition to protein and nucleic acid.Modern pharmacological research table
Bright polysaccharide has a variety of physiologically actives, including anti-oxidant, antitumor, antiviral, reducing blood lipid, anti-aging, enhancing immunologic function
Deng.And the polysaccharide in most of higher plant sources is the material having no adverse reaction, larger side effect will not be produced to body,
Therefore, the polysaccharide separated from plant causes great concern in biomedicine.Over one hundred kind of plant polyose has been carried out at present
Active related research report.Research shows that plant polyose can be combined by a variety of acceptors with immunocyte surface, activated not
With signal path regulate and control the immune system in animal body, including：Stimulating expression of macrophage, T/B lymphocytes, NKT
The secretion of cell or propagation；Adjust the release of cell factor；Promote the secretion of antibody；Activating complement system etc..
The mechanism of action of these active materials is also evolving, wherein immunologic mechanism of the polysaccharide to non-specificity induction
More it is valued by people.Plant polyose is optimal vaccine candidate medicine.
Tree butterfly is ascolichen class (Ascolichenes) Lecanorales (Lecanorales) ox-hide leaf section
(Stictaceae) Lobaria (Lobaria Hoffm) Plant Light oakmoss, split the drying foliaceous thallus of bud oakmoss and retigera Lobaria,
With good pharmacological action, there is invigorating spleen for diuresis, dispelling wind and arresting itching and other effects.Almost do not have to the research report for setting butterfly polysaccharide
Have.Most of natural activity polysaccharide are the natural green products having no toxic side effect.With deepening continuously for research, natural polysaccharide is got over
More to turn into food, health products, the study hotspot of medicine and other fields.
The content of the invention
For overcome the deficiencies in the prior art and shortcoming, primary and foremost purpose of the invention is to provide a kind of tree butterfly polysaccharide
Preparation method.
Another object of the present invention is to provide the tree butterfly polysaccharide being prepared by above-mentioned preparation method, obtain its polysaccharide
Yield is higher.
It is still another object of the present invention to provide the application of above-mentioned tree butterfly polysaccharide.
The purpose of the present invention is realized by following proposal：
A kind of preparation method for setting butterfly polysaccharide, is comprised the following steps：
(1) butterfly grinding and sieving will be set, obtains setting butterfly coarse powder；Distilled water is added in butterfly coarse powder to setting, at ultrasound
Reason, heating and refluxing extraction, filtering, and filtrate is concentrated, the tree butterfly Thick many candies solution concentrated；
(2) butterfly Thick many candies solution will be set and the macroporous absorbent resin by pre-processing mixes, decolorization, filtered out more
Sugar juice, and wash out the polysaccharide adsorbed in resin repeatedly with water, the polysaccharide solution after decolouring is merged, is concentrated under reduced pressure, obtains dense
Contracting liquid glucose；
(3) add precipitating reagent into concentration liquid glucose to be precipitated, centrifuge, collect sediment and drying, it is thick to obtain tree butterfly
Polysaccharide.
Ultrasonic time described in step (1) is 10~20min；Ultrasonic power is 60~400W；
Step (1) temperature being heated to reflux is 75~100 DEG C；The time of step (1) described heating and refluxing extraction is 2
~4h；The number of heating and refluxing extraction described in step (1) is 2~4 times；Butterfly coarse powder and distilled water are set described in step (1)
Mass ratio be 1:20~1:30；When hot water return extraction is multiple, the quality of tree butterfly coarse powder and the distilled water used each time
Than for 1:20~1:30.
To be concentrated under reduced pressure, the temperature of concentration is 40~60 DEG C for concentration described in step (1)；Decompression is dense described in step (2)
The temperature of contracting is 40~60 DEG C.
The condition of step (2) described decolorization is in 40~60 DEG C of 2~4h of water-bath；
Macroporous absorbent resin described in step (2) is D354FD resins；
The method of the pretreatment of macroporous absorbent resin is described in step (2)：By macroporous absorbent resin successively using distillation
Water, salt acid soak, are washed to neutrality, then are soaked with sodium hydroxide solution, then are washed to neutrality, obtain the macropore by pretreatment
Polymeric adsorbent.The time of the distilled water immersion is 12~24h, and the concentration of hydrochloric acid is 3~5wt%, and the time of salt acid soak is
0.5~3h, the concentration of sodium hydroxide solution is 3~5wt%, and the time of sodium hydroxide solution immersion is 0.5~3h.
The ethanol solution that precipitating reagent is absolute ethyl alcohol or volume fraction is 90~100% described in step (3)；
The temperature precipitated described in step (3) is 0~4 DEG C.
The addition of precipitating reagent described in step (3) is 3~5 times of concentration liquid glucose volume；Precipitated described in step (3)
Time is 8~24h；
The rotating speed centrifuged described in step (3) is 3000~5000r/min；The time of the centrifugation is 12~15min；Step
Suddenly the temperature dried described in (3) is 40~60 DEG C, is dried to constant weight.
One kind tree butterfly polysaccharide (LKY), is prepared by above-mentioned preparation method.
Application of the tree butterfly polysaccharide in anticancer, anti-oxidant and/or medicament for immunity enhancement is prepared.
The tree butterfly Thick many candies can be as new anticancer, anti-oxidant, medicament for immunity enhancement.
The principle of the present invention：The polysaccharide of plant origin has a variety of biologies such as anti-oxidant, antitumor and enhancing immunologic function
Activity.Polysaccharide especially can embody various beneficial pharmacological actions by adjusting the immunologic function of macrophage.The present invention passes through
Experiment in vitro is found and confirmation plant Thick many candies have anti-oxidant, antitumor and immune-enhancing activity, and dosage is low, same
Toxic side effect under sample dosage is relatively low.
The present invention is had the following advantages relative to prior art and effect：
(1) tree butterfly polysaccharide (LKY) of the invention is to human breast cancer cell line Bcap-37, lung carcinoma cell HCC827 and liver cancer
HepG-2 has certain inhibited proliferation；
(2) tree butterfly polysaccharide (LKY) of the invention has humidification to the release for strengthening RAW264.7 macrophages NO；
(3) tree butterfly polysaccharide (LKY) of the invention can significantly increase RAW264.7 macrophages IL-6 release；
(4) tree butterfly polysaccharide (LKY) of the invention produces TNF-α release to RAW264.7, and there is more significant enhancing to make
With.
Brief description of the drawings
Fig. 1 is effects of the tree butterfly polysaccharide LKY to the proliferation inhibition rate of human breast cancer cell line Bcap-37 of the preparation of embodiment 1
Figure；
Fig. 2 is design sketch of the tree butterfly polysaccharide LKY to lung carcinoma cell HCC827 proliferation inhibition rate of the preparation of embodiment 1；
Fig. 3 is design sketch of the tree butterfly polysaccharide LKY to lung carcinoma cell HepG-2 proliferation inhibition rate of the preparation of embodiment 1；
Fig. 4 is design sketch of the tree butterfly polysaccharide LKY to RAW264.7 macrophage survival rates of the preparation of embodiment 1；
Fig. 5 is the influence figure that tree butterfly polysaccharide LKY prepared by embodiment 1 discharges NO to RAW264.7 macrophages；Its
In, *：Compared with control groups (Negative control), P<0.05；**：Compared with control groups, P<0.01；
Fig. 6 is the influence figure that tree butterfly polysaccharide LKY prepared by embodiment 1 discharges IL-6 to RAW264.7 macrophages；Its
In, *：Compared with control groups (Negative control), P<0.05；**：With control groups (Negative
Control) compare, P<0.01；
Fig. 7 is the influence figure that tree butterfly polysaccharide LKY prepared by embodiment 1 discharges TNF-α to RAW264.7 macrophages；Its
In, *：Compared with control groups (Negative control), P<0.05；**：With control groups (Negative
Control) compare, P<0.01.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.In embodiment, human breast cancer cell line Bcap-37, lung carcinoma cell HCC827, HepG-2 cell and mouse macrophage
RAW 264.7 is purchased from Shanghai life science institute of Chinese Academy of Sciences cell resource center；Tree butterfly is purchased from the peaceful medicinal material market in Guangzhou.
Tree butterfly polysaccharide (LKY) is prepared from tree butterfly in embodiment 1：
(1) set after butterfly crushes and cross 20 mesh sieves, weigh 100g, add distilled water, 20min is handled with 60W power ultrasonic,
Heating and refluxing extraction, wherein, ratio of water to material 20:1 (mass ratio), refluxing extraction temperature are 75 DEG C, reflux extracting time 4h, are returned
Flow extraction time 3 times；Extract solution vacuum filtration is then combined with, filtrate is concentrated under reduced pressure at 40 DEG C with Rotary Evaporators
200mL, obtain polysaccharide solution；
(2) D354FD resins are first used into distilled water immersion 12h, then soaked with the hydrochloric acid solution that mass fraction is 5%
0.5h, distillation is washed to neutrality, then soaks 0.5h with the sodium hydroxide solution that mass fraction is 5%, and distillation is washed to neutrality,
200 mesh filtered through gauze, obtain the D354FD resins by pretreatment；
(3) D354FD resin mixing of the polysaccharide solution and 200g step (1) being prepared by pretreatment, is placed in 50
Water bath with thermostatic control 3h is decolourized in DEG C thermostat water bath, interval stirring, then filters out polysaccharide solution, and clean (5 repeatedly with water
It is secondary) go out the polysaccharide adsorbed in resin, the polysaccharide liquid after decolouring is merged, is concentrated under reduced pressure into Rotary Evaporators at 40 DEG C
200mL, obtain concentrating liquid glucose；
(4) under conditions of stirring, the absolute ethyl alcohol of 4 times of volumes is added in the concentration liquid glucose to step (3), in 4 DEG C of bars
Alcohol precipitation 8h under part, 15min is then centrifuged under 4000rpm, abandoning supernatant, sediment is taken out in 45 DEG C of drying, is set
Butterfly polysaccharide (LKY).
Tree butterfly polysaccharide (LKY) is prepared from tree butterfly in embodiment 2：
(1) set after butterfly crushes and cross 20 mesh sieves, weigh 100g, add distilled water, 15min is handled with 80W power ultrasonic,
Heating and refluxing extraction, ratio of water to material 20:1 (mass ratio), refluxing extraction temperature are 80 DEG C, reflux extracting time 3h, refluxing extraction
Number 2 times；Extract solution is then combined with, is filtered by vacuum, filtrate at 50 DEG C is concentrated under reduced pressure into 200mL with Rotary Evaporators, obtained
To polysaccharide solution；
(2) by D354FD resins distilled water immersion 24h, 2h then is soaked with the hydrochloric acid solution that mass fraction is 5%, is steamed
Distilled water is washed till neutrality, then soaks 2h with the sodium hydroxide solution that mass fraction is 5%, and distillation is washed to neutrality, 200 mesh gauze mistakes
Filter, obtains the D354FD resins by pretreatment；
(3) D354FD resin mixing of the polysaccharide solution and 200g step (1) being prepared by pretreatment, is placed in 40
Water bath with thermostatic control 2h is decolourized in DEG C thermostat water bath, interval stirring, then filters out polysaccharide solution, and clean (4 repeatedly with water
It is secondary) go out the polysaccharide adsorbed in resin, the polysaccharide liquid after decolouring is merged, is concentrated under reduced pressure into Rotary Evaporators at 50 DEG C
200mL, obtain concentrating liquid glucose；
(4) under conditions of stirring, the volume fraction that 3 times of volumes are added in the concentration liquid glucose to step (3) is 95%
Ethanol solution, the alcohol precipitation 12h under the conditions of 0 DEG C, 12min is then centrifuged under 3000rpm, abandoning supernatant, takes out sediment
In 50 DEG C of drying, obtain setting butterfly polysaccharide (LKY).
Tree butterfly polysaccharide (LKY) is prepared from tree butterfly in embodiment 3：
(1) set after butterfly crushes and cross 20 mesh sieves, weigh 100g, add distilled water, handled with 100W power ultrasonic
10min, heating and refluxing extraction, wherein, ratio of water to material 20:1 (mass ratio), refluxing extraction temperature are 95 DEG C, reflux extracting time
For 2h, refluxing extraction number 4 times, extract solution is then combined with, be filtered by vacuum, filtrate depressurized at 60 DEG C with Rotary Evaporators dense
200mL is reduced to, obtains polysaccharide solution；
(2) by D354FD resins distilled water immersion 18h, 3h then is soaked with the hydrochloric acid solution that mass fraction is 5%, is steamed
Distilled water is washed till neutrality, then soaks 3h with the sodium hydroxide solution that mass fraction is 5%, and distillation is washed to neutrality, 200 mesh gauze mistakes
Filter, obtains the D354FD resins by pretreatment；
(3) the D354FD resins of the polysaccharide solution and 200g that are prepared in step (1) by pretreatment are mixed, be placed in
Water bath with thermostatic control 4h is decolourized in 60 DEG C of thermostat water baths, interval stirring, goes out polysaccharide solution with the filter-cloth filtering of 200 mesh afterwards,
And clean (5 times) repeatedly with water and wash out the polysaccharide adsorbed in resin, the polysaccharide liquid after decolouring is merged, with Rotary Evaporators 60
200mL is concentrated under reduced pressure at DEG C, obtains concentrating liquid glucose；
(4) under conditions of stirring, the volume fraction that 5 times of volumes are added in the concentration liquid glucose to step (3) is 95%
Ethanol, the alcohol precipitation 24h under the conditions of 4 DEG C, 14min is then centrifuged under 5000rpm, abandoning supernatant, takes out sediment in 60
DEG C drying, obtain set butterfly polysaccharide (LKY).
The tree butterfly polysaccharide (LKY) of embodiment 4 is to human breast cancer cell line Bcap-37 and lung carcinoma cell HCC827, liver cancer cells
HepG-2 inhibited proliferation
By human breast cancer cell line Bcap-37 and lung carcinoma cell HCC827 cells, HepG-2 cell, with 1 × 105Individual/ml
Concentration is inoculated in 96 porocyte culture plates (100 μ L/ holes), puts 37 DEG C, is cultivated in the CO2gas incubator of volume fraction 5%.Treat
After cell in orifice plate is completely adherent, the sample solution 100 of various concentrations (125,250,500,1000,2000 μ g/m L) is added
μ l (sample solution that tree butterfly polysaccharide prepared by embodiment 1 is made into).Each concentration set 6 it is parallel.At 37 DEG C, 5%CO2Culture
Continue to cultivate in case, taken out after 24h, inverted microscope observes the change of cellular morphology.Carefully inhaled and abandoned in 96 orifice plates with syringe
Nutrient solution, with the PBS board-washings 2 times of not calcic, magnesium ion, 5mg/m L μ L of MTT solution 20 are then added per hole, and (need to turn off the light behaviour
Make, MTT is shown in that light easily decomposes) and μ L of DMEM complete culture solutions 180, at 37 DEG C, 5%CO2Continue to cultivate 4h in incubator.It is careful to inhale
The cell culture fluid abandoned in hole, 150 μ L DMSO are added per hole, vibrate 10min.In wavelength it is 490nm with enzyme-linked immunosorbent assay instrument
Lower its absorbance of measure.
The proliferation inhibition rate of cancer cell=(blank control group absorbance-sample sets absorbance)/blank control group
Light absorption value.Blank control group is complete medium.
Test result as shown in Figure 1, Figure 2 and Figure 3；Fig. 1 is the tree butterfly polysaccharide (LKY) of the preparation of embodiment 1 to people's mammary gland
The design sketch of cancer cell MCF-7 proliferation inhibition rate；Fig. 2 is the tree butterfly polysaccharide (LKY) of the preparation of embodiment 1 to lung carcinoma cell
The design sketch of HCC827 proliferation inhibition rate；Fig. 3 is the tree butterfly polysaccharide (LKY) of the preparation of embodiment 1 to HepG-2 cell
Proliferation inhibition rate design sketch.
Tree butterfly polysaccharide (LKY) prepared by embodiment 1 increases with the increase of concentration the inhibiting rate of cancer cell；Set butterfly
Polysaccharide LKY has certain rejection ability to HCC827, HepG-2；In summary, tree butterfly purified polysaccharide LKY has antitumor work
Property.
Influences of the tree butterfly polysaccharide LKY of embodiment 5 to RAW264.7 cell survival rates
By mouse monokaryon macrophage RAW264.7 with 1 × 106Individual/ml concentration is inoculated in 96 porocyte culture plates (100 μ
L/ holes), 37 DEG C are put, 5%CO2Cultivated in incubator.After the cell in orifice plate is completely adherent, 100 μ L various concentrations are added
Tree butterfly polysaccharide (31.25,62.5125,250,500,1000 μ g/mL) prepared by embodiment 1, while set cell blank control
Group.At 37 DEG C, 5%CO2Continue to cultivate in incubator, taken out after 24h, the metamorphosis of cell is observed under inverted microscope.With
Syringe, which is carefully inhaled, abandons culture supernatants in hole, and cell survival rate detection is carried out with twice of PBS, then with mtt assay, i.e. every hole
The 5mg/m L μ L of 20 μ L and DMEM complete culture solution of MTT solution 180 are added, at 37 DEG C, 5%CO2Continue to cultivate in incubator
4h.Then the supernatant abandoned in hole is carefully inhaled, 150 μ L DMSO is added per hole, vibrates 10min.Existed with enzyme-linked immunosorbent assay instrument
Wavelength is to determine its absorbance under 490nm, and the survival rate of cell is calculated according to formula below:
Relative propagation degree (PGR)=experimental group mean absorbance values/blank control group absorbance × 100%.Experimental group
As add the experimental group of tree butterfly polysaccharide.
Test result is as shown in Figure 4；Fig. 4 is that tree butterfly polysaccharide LKY prepared by embodiment 1 deposits to RAW264.7 macrophages
The influence figure of motility rate.
As a result show：Tree butterfly polysaccharide of the concentration below 500 μ g/mL to RAW264.7 cells without overt toxicity, however,
When polysaccharide concentration is more than 1000 μ g/mL, generation overt toxicity.Therefore, polysaccharide sample is in 31.25~500 μ g/mL concentration ranges
The immunocompetent research of subsequent cell can be carried out.
The Griess methods of embodiment 6 detect the influence that LKY discharges NO to RAW264.7 cells
Cellar culture cell, the RAW264.7 cells in growth period of taking the logarithm, piping and druming cell is into single cell suspension, under microscope
After being counted with blood cell counting plate, 1000r/min, centrifugation 5min removes supernatant, and culture medium is resuspended and adjusts cell concentration, by 200,000
The cell density in individual/hole is inoculated in 24 well culture plates, is placed in 37 DEG C, 5%CO2Incubator culture, adherent 24h, suction are abandoned supernatant, pressed
Following packet requires administration, and every group sets 3 multiple holes：Control groups (blank group), lipopolysaccharides (LPS, final concentration of 1 μ g/mL)
Group, polysaccharide sample group (the tree butterfly polysaccharide LKY (μ of final concentration of 31.25,62.5,125,250 and 500 that embodiment 1 is prepared
G/mL), the culture medium of control group addition same volume.After administration, 37 DEG C are placed in, 5%CO2Incubator kind, continue to cultivate 24h.Point
Each μ L of hole cell conditioned medium 100 are not taken, are accordingly added in another 96 new orifice plates.Add 50 μ L Griess reagent As and 50 μ L per hole
Griess reagent B, be subsequently placed in 37 DEG C of incubators and react 10min, put plate immediately on multiple labeling micropore board detector,
Each hole absorbance is detected under 550nm wavelength.
Test result is as shown in Figure 5；Fig. 5 is that tree butterfly polysaccharide LKY prepared by embodiment 1 is released RAW264.7 macrophages
Put NO influence figure.Tree butterfly polysaccharide has the function that promotion RAW264.7 produces NO, and in 31.25~500 μ g/mL concentration models
In enclosing, NO burst size increases and increased with concentration.
The Elisa methods of embodiment 7 detection tree butterfly polysaccharide LKY discharges the influence of IL-6 and TNF-α to RAW264.7 cells
Cellar culture cell, the RAW264.7 cells in growth period of taking the logarithm, piping and druming cell is into single cell suspension, under microscope
After being counted with blood cell counting plate, 1000r/min, centrifugation 5min removes supernatant, and culture medium is resuspended and adjusts cell concentration, by 200,000
The cell density in individual/hole is inoculated in 24 well culture plates, is placed in 37 DEG C, 5%CO2Incubator culture, adherent 24h, suction are abandoned supernatant, pressed
Following packet requires administration, and every group sets 3 multiple holes：Control groups (blank group), lipopolysaccharides (LPS, final concentration of 1 μ g/mL)
Group, LKY groups (LKY that embodiment 1 is prepared, final concentration of 31.25,62.5,125,250,500 μ g/mL), control group addition
The culture medium of same volume.After administration, 37 DEG C are placed in, 5%CO2In incubator, continue to cultivate 24h.Each hole cell conditioned medium is taken respectively
According to mouse IL-6ELISA kits and the release conditions of mouse TNF-α ELISA kit operating method detection cell factor.
Test result is as shown in Figure 6,7；Fig. 6 is the tree butterfly polysaccharide LKY of the preparation of embodiment 1 to RAW264.7 macrophages
Discharge IL-6 influence figure.Fig. 7 is that tree butterfly polysaccharide LKY prepared by embodiment 1 discharges TNF-α to RAW264.7 macrophages
Influence figure.
As a result show, in 31.25~500 μ g/mL concentration ranges, tree butterfly polysaccharide LKY to RAW264.7 produce IL-6,
TNF-α has certain effect, and acts on enhancing with polysaccharide concentration increase, all has pole conspicuousness (p under each concentration<0.01).
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
- A kind of 1. preparation method for setting butterfly polysaccharide, it is characterised in that：Comprise the following steps：(1) butterfly grinding and sieving will be set, obtains setting butterfly coarse powder；Distilled water is added into tree butterfly coarse powder, is ultrasonically treated, adds Circumfluence distillation, filtering, and filtrate is concentrated, the tree butterfly Thick many candies solution concentrated；(2) butterfly Thick many candies solution will be set and the macroporous absorbent resin by pre-processing mixes, decolorization, it is molten to filter out polysaccharide Liquid, and wash out the polysaccharide adsorbed in resin repeatedly with water, the polysaccharide solution after decolouring is merged, is concentrated under reduced pressure, obtains concentration sugar Liquid；(3) add precipitating reagent into concentration liquid glucose to be precipitated, centrifuge, collect sediment and drying, obtain setting butterfly Thick many candies.
- 2. the preparation method of butterfly polysaccharide is set according to claim 1, it is characterised in that：The condition of step (2) described decolorization is in 40~60 DEG C of 2~4h of water-bath；The ethanol solution that precipitating reagent is absolute ethyl alcohol or volume fraction is 90~100% described in step (3)；The temperature of the precipitation Spend for 0~4 DEG C.
- 3. the preparation method of butterfly polysaccharide is set according to claim 1, it is characterised in that：Ultrasonic power described in step (1) For 60~400W, ultrasonic time is 10~20min；The temperature being heated to reflux is 75~100 DEG C；Step heats back described in (1) The time of stream extraction is 2~4h；The number of the heating and refluxing extraction is 2~4 times；The mass ratio that butterfly coarse powder and distilled water are set described in step (1) is 1:20~1:30；When hot water return extraction is multiple, tree The mass ratio of butterfly coarse powder and the distilled water used each time is 1:20~1:30.
- 4. the preparation method of butterfly polysaccharide is set according to claim 1, it is characterised in that：Macroporous absorption described in step (2) Resin is D354FD resins.
- 5. the preparation method of butterfly polysaccharide is set according to claim 1, it is characterised in that：Macroporous absorption described in step (2) The method of the pretreatment of resin is：By macroporous absorbent resin successively using distilled water, salt acid soak, wash to neutrality, then use hydrogen Sodium hydroxide solution is soaked, then is washed to neutrality, obtains the macroporous absorbent resin by pretreatment.
- 6. the preparation method of butterfly polysaccharide is set according to claim 5, it is characterised in that：The time of the distilled water immersion is 12~24h, the concentration of hydrochloric acid are 3~5wt%, and time of salt acid soak is 0.5~3h, the concentration of sodium hydroxide solution for 3~ 5wt%, the time of sodium hydroxide solution immersion is 0.5~3h.
- 7. the preparation method of butterfly polysaccharide is set according to claim 1, it is characterised in that：Precipitating reagent described in step (3) Addition is 3~5 times of concentration liquid glucose volume；The time precipitated described in step (3) is 8~24h；To be concentrated under reduced pressure, the temperature of concentration is 40~60 DEG C for concentration described in step (1)；It is concentrated under reduced pressure described in step (2) Temperature is 40~60 DEG C.
- 8. the preparation method of butterfly polysaccharide is set according to claim 1, it is characterised in that：What is centrifuged described in step (3) turns Speed is 3000~5000r/min；The time of the centrifugation is 12~15min；The temperature dried described in step (3) is 40~60 ℃。
- A kind of 9. tree butterfly polysaccharide obtained by any one of claim 1~8 preparation method.
- 10. the application of butterfly polysaccharide is set according to claim 9, it is characterised in that：It is described tree butterfly polysaccharide prepare anticancer, Application in anti-oxidant and/or medicament for immunity enhancement.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106478833A (en) * | 2016-11-09 | 2017-03-08 | 华南理工大学 | A kind of Calyx Hibisci Sabdariffae polysaccharide and preparation method and application |
| CN106519055A (en) * | 2016-11-11 | 2017-03-22 | 华南理工大学 | Bitter orange calyx polysaccharide and preparation method and application thereof |
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2017
- 2017-09-01 CN CN201710778774.0A patent/CN107586346A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106478833A (en) * | 2016-11-09 | 2017-03-08 | 华南理工大学 | A kind of Calyx Hibisci Sabdariffae polysaccharide and preparation method and application |
| CN106519055A (en) * | 2016-11-11 | 2017-03-22 | 华南理工大学 | Bitter orange calyx polysaccharide and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
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| 申超群等: "《树蝴蝶多糖LKY-I的结构和抗氧化、抗肿瘤活性评价》", 《食品科学》 * |
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