CN107868136A - A kind of extracting method of the Chinese pholidota pseudobulb or herb polysaccharide with cancer suppressing action - Google Patents
A kind of extracting method of the Chinese pholidota pseudobulb or herb polysaccharide with cancer suppressing action Download PDFInfo
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- CN107868136A CN107868136A CN201711094188.0A CN201711094188A CN107868136A CN 107868136 A CN107868136 A CN 107868136A CN 201711094188 A CN201711094188 A CN 201711094188A CN 107868136 A CN107868136 A CN 107868136A
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- 241000283966 Pholidota <mammal> Species 0.000 title claims abstract description 38
- 150000004676 glycans Chemical class 0.000 title claims abstract description 23
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 23
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 20
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 8
- 201000011510 cancer Diseases 0.000 title claims abstract description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 52
- 238000001556 precipitation Methods 0.000 claims abstract description 22
- 239000000287 crude extract Substances 0.000 claims abstract description 19
- 238000000605 extraction Methods 0.000 claims abstract description 9
- 239000012153 distilled water Substances 0.000 claims description 35
- 238000000502 dialysis Methods 0.000 claims description 16
- 239000006228 supernatant Substances 0.000 claims description 15
- 238000005119 centrifugation Methods 0.000 claims description 12
- 235000019441 ethanol Nutrition 0.000 claims description 11
- 239000000284 extract Substances 0.000 claims description 11
- 239000012141 concentrate Substances 0.000 claims description 10
- 239000012535 impurity Substances 0.000 claims description 10
- 239000000463 material Substances 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 9
- 125000005909 ethyl alcohol group Chemical group 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 7
- 238000004108 freeze drying Methods 0.000 claims description 6
- 229920002271 DEAE-Sepharose Polymers 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- 239000011343 solid material Substances 0.000 claims description 5
- 230000001186 cumulative effect Effects 0.000 claims description 4
- 238000007667 floating Methods 0.000 claims description 4
- 230000014759 maintenance of location Effects 0.000 claims description 4
- 239000004575 stone Substances 0.000 claims description 4
- 238000000746 purification Methods 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims 2
- 230000008014 freezing Effects 0.000 claims 2
- 235000006041 Prunus persica f compressa Nutrition 0.000 claims 1
- 240000006522 Prunus persica f. compressa Species 0.000 claims 1
- 206010009944 Colon cancer Diseases 0.000 abstract description 4
- 208000029742 colonic neoplasm Diseases 0.000 abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 3
- 241000238366 Cephalopoda Species 0.000 abstract description 2
- 238000007445 Chromatographic isolation Methods 0.000 abstract description 2
- 238000011097 chromatography purification Methods 0.000 abstract description 2
- 238000004134 energy conservation Methods 0.000 abstract description 2
- 239000012467 final product Substances 0.000 abstract description 2
- 238000010257 thawing Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 16
- 239000000499 gel Substances 0.000 description 9
- 238000010828 elution Methods 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000011521 glass Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 238000002390 rotary evaporation Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 206010011224 Cough Diseases 0.000 description 1
- 240000007817 Olea europaea Species 0.000 description 1
- 241000233855 Orchidaceae Species 0.000 description 1
- 241001048892 Pholidota chinensis Species 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007596 consolidation process Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Materials Engineering (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Sustainable Development (AREA)
- Medicines Containing Plant Substances (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of extracting method of the Chinese pholidota pseudobulb or herb polysaccharide with cancer suppressing action, water-swollen squid is carried out to Chinese pholidota pseudobulb or herb polysaccharide using the extracting method of the present invention, including by Chinese pholidota pseudobulb or herb water bath with thermostatic control, alcohol precipitation freeze thawing, obtain the first crude extract, de- albumen is carried out to the first crude extract again and obtains the second crude extract, then column chromatographic isolation and purification, obtain final product, above method extraction Chinese pholidota pseudobulb or herb polysaccharide is time saving, energy-conservation, and the yield of purified polysaccharide is high, the polysaccharide of extraction has the function that preferably to suppress Growth of Colon Cancer Cells.
Description
Technical field
The invention belongs to technical field of natural product extraction, and in particular to a kind of Chinese pholidota pseudobulb or herb polysaccharide with cancer suppressing action
Extracting method.
Background technology
Chinese pholidota pseudobulb or herb (Pholidota chinensis Lindl) wears disk, stone an enclosure for storing grain meat also referred to as stone olive, Shi Shanglian, stone
Deng it is a kind of common medicinal orchid family Pholidota plant, and medicinal position is pseudobulb or herb, in being mainly distributed on
The state southeast.The bulb of Chinese pholidota pseudobulb or herb has anti-inflammatory, analgesia and the effect of cough-relieving as its major part.Have in Chinese pholidota pseudobulb or herb bulb
Various bioactivators, found through this laboratory research, Chinese pholidota pseudobulb or herb polysaccharide, which has, suppresses human colon cancer cell caco-2 propagation
Effect, therefore be necessary to further investigate for the extracting method of Chinese pholidota pseudobulb or herb bulb polysaccharide.
The content of the invention
It is an object of the invention to overcome prior art defect, there is provided a kind of Chinese pholidota pseudobulb or herb polysaccharide with cancer suppressing action carries
Take method.
Technical scheme is as follows：
A kind of extracting method of the Chinese pholidota pseudobulb or herb polysaccharide with cancer suppressing action, comprises the following steps：
(1) Chinese pholidota pseudobulb or herb bulb portion is taken, after fully drying, dry Chinese pholidota pseudobulb or herb bulb sample is weighed and is put into distilled water,
The mass ratio of the Chinese pholidota pseudobulb or herb bulb sample and distilled water is 1: 28~32, and 2.5~4h is extracted in 92~95 DEG C of water-baths of thermostat water bath,
3000~4000r/min centrifuges 4~6min, goes the removal of impurity, obtains extract solution；
(2) extract solution obtained by step (1) is concentrated for 65~75 DEG C with Rotary Evaporators, after solution to be concentrated is cooled to room temperature
3~5 times of volume absolute ethyl alcohols are added, is placed in 3~5 DEG C of refrigerators and preserves more than 10h, 3000~4000r/min centrifugations 4~
6min, supernatant is abandoned, take precipitation；
(3) 4~6min is centrifuged with the precipitation obtained by distilled water dissolving step (2), multigelation, 3000~4000r/min
Impurity is removed, vacuum freeze drier drying is subsequently placed in, obtains the first crude extract, the condition of the multigelation of the step is：
Cryogenic temperature is -22~-18 DEG C, and solution temperature is 45~55 DEG C, is repeated 4~6 times；
(4) take above-mentioned first crude extract to weigh, be completely dissolved according to 1: 24~27 mass ratio in distilled water, by totality
Product 1/4 adds Sevage reagents, fully vibrates 1.5~2.5h, and 3000~4000r/min centrifuges 8~15min, and material divides three layers,
Intermediate layer is solid material, collects supernatant liquor, then the material in intermediate layer and lower floor is repeated the step handled to
Untill intermediate layer is without solid-like floating preteins, concentrate is concentrated to give to above-mentioned supernatant liquor with Rotary Evaporators, in the concentrate
3~5 times of volume absolute ethyl alcohols of middle addition, 3~5 DEG C stand overnight, and then 3000~4000r/min centrifuges 4~6min centrifuging and takings
Precipitation；
(5) precipitation obtained by distilled water dissolving step (4) is used, is concentrated after multigelation with Rotary Evaporators, using retention
Bag filter flowing water 20~25h of dialysis that molecular weight is 3300~3800, distilled water 20~25h of dialysis, freeze-drying obtain second
Crude extract, the condition of the multigelation of the step are：Cryogenic temperature is -22~-18 DEG C, and solution temperature is 45~55 DEG C, repeats 4
~6 times；
(6) the second runic thing is carried out after purification with DEAE-Sepharose CL-6B chromatographic columns, then by dialysis and it is cold
It is lyophilized dry, obtain the Chinese pholidota pseudobulb or herb polysaccharide.
In a preferred embodiment of the invention, the step (1) is：Chinese pholidota pseudobulb or herb bulb portion is taken, is fully dried
Afterwards, weigh dry Chinese pholidota pseudobulb or herb bulb sample to be put into distilled water, the mass ratio of the Chinese pholidota pseudobulb or herb bulb sample and distilled water is 1:
30,95 DEG C of water-bath extraction 3h, 3500r/min centrifugation 5min of thermostat water bath, the removal of impurity is gone, obtains extract solution.
In a preferred embodiment of the invention, the step (2) is：Extract solution obtained by step (1) is revolved
Turn 70 DEG C of concentrations of evaporimeter, solution to be concentrated adds 4 times of volume absolute ethyl alcohols after being cooled to room temperature, is placed in 4 DEG C of refrigerators and preserves 10h
More than, 3500r/min centrifugation 5min, supernatant is abandoned, takes precipitation.
In a preferred embodiment of the invention, the step (3) is：Obtained by distilled water dissolving step (2)
Precipitation, multigelation, 3500r/min centrifugations 5min remove impurity, are subsequently placed in vacuum freeze drier drying, it is thick to obtain first
Extract, the condition of the multigelation of the step are：Cryogenic temperature is -20 DEG C, and solution temperature is 50 DEG C, is repeated 5 times.
In a preferred embodiment of the invention, the step (4) is：Above-mentioned first crude extract is taken to weigh, according to 1
: 25 mass ratio is completely dissolved in distilled water, is added Sevage reagents by cumulative volume 1/4, is fully vibrated 2h, 3500r/min
10min is centrifuged, three layers of material point, intermediate layer is solid material, supernatant liquor is collected, then to intermediate layer and the material of lower floor
Repeat the step to be handled untill intermediate layer is without solid-like floating preteins, above-mentioned supernatant liquor is concentrated with Rotary Evaporators
Concentrate is obtained, 4 times of volume absolute ethyl alcohols are added in the concentrate, 4 DEG C stand overnight, and then 3500r/min centrifuges 5min and taken
Precipitation.
In a preferred embodiment of the invention, the step (5) is：Obtained by distilled water dissolving step (4)
Precipitate, concentrated after multigelation with Rotary Evaporators, use molecular cut off as 3500 bag filter flowing water dialysis 24h, distillation
Water dialysis 24h, freeze-drying obtain the second crude extract, and the condition of the multigelation of the step is：Cryogenic temperature is -20 DEG C, molten
It is 50 DEG C to solve temperature, is repeated 5 times.
In a preferred embodiment of the invention, the retention of the bag filter used in the dialysis in the step (6) point
Son amount is 3300~3800.
The beneficial effects of the invention are as follows：Water-swollen squid is carried out to Chinese pholidota pseudobulb or herb polysaccharide using the extracting method of the present invention, including
By Chinese pholidota pseudobulb or herb water bath with thermostatic control, alcohol precipitation freeze thawing, the first crude extract is obtained, then carry out de- albumen to the first crude extract and obtain second slightly carrying
Thing, then column chromatographic isolation and purification, obtains final product, above method extraction Chinese pholidota pseudobulb or herb polysaccharide is time saving, energy-conservation, and purified polysaccharide
Yield it is high, the polysaccharide of extraction has the function that preferably to suppress Growth of Colon Cancer Cells.
Embodiment
Technical scheme is further detailed and described below by way of embodiment.
Embodiment 1
A kind of extracting method of the Chinese pholidota pseudobulb or herb polysaccharide with cancer suppressing action, comprises the following steps：
(1) Chinese pholidota pseudobulb or herb bulb portion is taken, after fully drying, dry Chinese pholidota pseudobulb or herb bulb sample 100g is weighed and is put into 3000mL
In distilled water, 95 DEG C of water-bath extraction 3h, 3500r/min centrifugation 5min of thermostat water bath, the removal of impurity is gone, obtains extract solution；
(2) the 70 DEG C of concentrations of Rotary Evaporators of the extract solution obtained by step (1), solution to be concentrated are added after being cooled to room temperature
4 times of volume absolute ethyl alcohols, it is placed in 4 DEG C of refrigerators and preserves more than 10h, 3500r/min centrifugation 5min, abandons supernatant, take precipitation；
(3) impurity is removed with the precipitation obtained by distilled water dissolving step (2), multigelation, 3500r/min centrifugations 5min,
Vacuum freeze drier drying is subsequently placed in, obtains the first crude extract, the condition of the multigelation of the step is：Cryogenic temperature
For -20 DEG C, solution temperature is 50 DEG C, is repeated 5 times；
(4) take above-mentioned first crude extract to weigh, be completely dissolved according to 1: 25 mass ratio in distilled water, by cumulative volume 1/
4 add Sevage reagents, fully vibrate 2h, and 3500r/min centrifugation 10min, three layers of material point, intermediate layer is solid material,
Supernatant liquor is collected, the step is then repeated to the material in intermediate layer and lower floor is handled to intermediate layer to dissociate egg without solid-like
Untill white, concentrate is concentrated to give to above-mentioned supernatant liquor with Rotary Evaporators, 4 times of anhydrous second of volume are added in the concentrate
Alcohol, 4 DEG C stand overnight, and then 3500r/min centrifuges 5min and takes precipitation；
(5) precipitation obtained by distilled water dissolving step (4) is used, is concentrated after multigelation with Rotary Evaporators, using retention
The bag filter flowing water dialysis 24h that molecular weight is 3500, distilled water dialysis 24h, freeze-drying obtain the second crude extract, the step
The condition of multigelation is：Cryogenic temperature is -20 DEG C, and solution temperature is 50 DEG C, is repeated 5 times；
(6) the second runic thing is carried out after purification with DEAE-Sepharose CL-6B chromatographic columns, then by dialysis and it is cold
It is lyophilized dry, the Chinese pholidota pseudobulb or herb polysaccharide is obtained, it is specific as follows：
A, the pretreatment of DEAE-Sepharose CL-6B gels：Gel is placed in vacuum desiccator, connects recirculated water
Formula vavuum pump, carry out interval and vacuumize, pumping 30min (interval 5min), and stirred with glass bar, make it loose, make in gel
Gas can be completely drawn out, then fill post, during post is filled should from top end import level pad (distilled water), with except
The air gone in dead angle, after removal of bubbles, distilled water is kept 3cm height in post, gel is gently stirred evenly with glass bar and is hanged
Supernatant liquid, gel suspension is drained into post by glass bar, gel suspension is added from post opening's edge inwall, avoid producing bubble；
Fill column diameter：4.6cm, height：40cm；Gel precipitation is treated, opens constant flow pump, flow velocity 60mL/h is adjusted, with 2000mL distilled water
Balance, makes gel consolidation；
B, sample detection and collection：The crude extracts of 500mg second are taken to be dissolved in 10mL distilled water, upper DEAE-Sepharose CL-
6B chromatographic columns (4.6 × 40cm), post upper strata buffer solution is suctioned out after maintaining an equal level with gel top layer, pipette samples solution is along chromatographic column
The rotation of wall same direction is slowly added to, and makes sample uniform in glue post surface distributed；After adding, constant flow pump is opened, enters sample liquid
Cylinder, when cylinder upper surface residue about 1mm sample liquids, along chromatography column wall same direction, rotation is slowly added to distilled water, will
Adhering liquid on wall is washed down, then proceedes to according to said method add distilled water to away from glue post upper surface about 8cm height；Cover cylinder；
Adjust flow velocity 60mL/h；Successively use respectively and distill water elution, 0~1M NaCl solution linear gradient elutions, flow velocity 1mL/min, if
Determine 12min and collect a pipe, often pipe 12mL, automatic fraction collector are collected；
C, sugared content distribution situation is determined with phend-sulphuric acid, is distributed with determined by ultraviolet spectrophotometry protein content
Situation, record data simultaneously draw elution curve, collect the purifying homogeneous components that sodium chloride linear gradient elution obtains, rotary evaporation
Concentration, with the bag filter of molecular cut off 3500 to collection liquid running water flowing water dialysis 24h, then distilled water dialysis 24h, dialyses
After the completion of carry out rotary evaporation concentration, freeze-drying, obtain the Chinese pholidota pseudobulb or herb polysaccharide.
To gained Chinese pholidota pseudobulb or herb polysaccharide system detectio its homogeneity of high performance liquid chromatography Agilent 1100, condition is：
Liquid-phase chromatographic column Shodex Sugar KS-804, flow velocity 1mL/min, temperature 50 C, Composition distribution.Pass through high-efficient liquid phase color
Spectrum detection, a homogeneity component (being named as PCP-I) can be obtained in 1mol/LNaCl linear gradient elutions.Using mtt assay
PCP-I active anticancer is detected, disclose PCP-I has obvious inhibitory action to human colon cancer cell caco-2 propagation, and it increases
It is 69.542ug/mL to grow inhibiting rate IC50 values.PCP-I belongs to natural extracts, and its source is to have long edible history
Chinese pholidota pseudobulb or herb, therefore there is extensive potentiality to be exploited.
The foregoing is only a preferred embodiment of the present invention, therefore can not limit the scope that the present invention is implemented according to this, i.e.,
The equivalent changes and modifications made according to the scope of the claims of the present invention and description, all should still it belong in the range of the present invention covers.
Claims (7)
- A kind of 1. extracting method of the Chinese pholidota pseudobulb or herb polysaccharide with cancer suppressing action, it is characterised in that：Comprise the following steps：(1) Chinese pholidota pseudobulb or herb bulb portion is taken, after fully drying, dry Chinese pholidota pseudobulb or herb bulb sample is weighed and is put into distilled water, the stone The mass ratio of flat peach bulb sample and distilled water is 1: 28~32, and 2.5~4h is extracted in 92~95 DEG C of water-baths of thermostat water bath, 3000 ~4000r/min centrifuges 4~6min, goes the removal of impurity, obtains extract solution；(2) 65~75 DEG C of concentrations of Rotary Evaporators of the extract solution obtained by step (1), solution to be concentrated are added after being cooled to room temperature 3~5 times of volume absolute ethyl alcohols, it is placed in 3~5 DEG C of refrigerators and preserves more than 10h, 3000~4000r/min centrifuges 4~6min, abandons Supernatant, take precipitation；(3) centrifuged 4~6min with the precipitation obtained by distilled water dissolving step (2), multigelation, 3000~4000r/min and removed Impurity, vacuum freeze drier drying is subsequently placed in, obtains the first crude extract, the condition of the multigelation of the step is：Freezing Temperature is -22~-18 DEG C, and solution temperature is 45~55 DEG C, is repeated 4~6 times；(4) take above-mentioned first crude extract to weigh, be completely dissolved according to 1: 24~27 mass ratio in distilled water, by cumulative volume 1/ 4 add Sevage reagents, fully vibrate 1.5~2.5h, and 3000~4000r/min centrifuges 8~15min, and three layers of material point is middle Layer is solid material, collects supernatant liquor, and then repeating the step to the material in intermediate layer and lower floor is handled to centre Untill layer is without solid-like floating preteins, concentrate is concentrated to give to above-mentioned supernatant liquor with Rotary Evaporators, added in the concentrate Enter 3~5 times of volume absolute ethyl alcohols, 3~5 DEG C stand overnight, and then 3000~4000r/min centrifuges 4~6min centrifuging and takings precipitation；(5) precipitation obtained by distilled water dissolving step (4) is used, is concentrated after multigelation with Rotary Evaporators, using retention molecule Bag filter flowing water 20~25h of dialysis for 3300~3800, distilled water 20~25h of dialysis are measured, freeze-drying obtains second and slightly carried Thing, the condition of the multigelation of the step are：Cryogenic temperature is -22~-18 DEG C, and solution temperature is 45~55 DEG C, repeats 4~6 It is secondary；(6) the second runic thing is carried out after purification with DEAE-Sepharose CL-6B chromatographic columns, then it is dry by dialysis and freezing It is dry, obtain the Chinese pholidota pseudobulb or herb polysaccharide.
- 2. extracting method as claimed in claim 1, it is characterised in that：The step (1) is：Chinese pholidota pseudobulb or herb bulb portion is taken, is filled Divide after drying, weigh dry Chinese pholidota pseudobulb or herb bulb sample and be put into distilled water, the quality of the Chinese pholidota pseudobulb or herb bulb sample and distilled water Than for 1: 30,95 DEG C of water-baths extraction 3h, 3500r/min centrifugation 5min of thermostat water bath, going the removal of impurity, obtaining extract solution.
- 3. extracting method as claimed in claim 1, it is characterised in that：The step (2) is：By the extraction obtained by step (1) Liquid 70 DEG C of concentrations of Rotary Evaporators, solution to be concentrated add 4 times of volume absolute ethyl alcohols after being cooled to room temperature, are placed in 4 DEG C of refrigerators More than 10h is preserved, 3500r/min centrifugation 5min, supernatant is abandoned, takes precipitation.
- 4. extracting method as claimed in claim 1, it is characterised in that：The step (3) is：With distilled water dissolving step (2) The precipitation of gained, multigelation, 3500r/min centrifugations 5min remove impurity, are subsequently placed in vacuum freeze drier drying, obtain First crude extract, the condition of the multigelation of the step are：Cryogenic temperature is -20 DEG C, and solution temperature is 50 DEG C, is repeated 5 times.
- 5. extracting method as claimed in claim 1, it is characterised in that：The step (4) is：Above-mentioned first crude extract is taken to claim Weight, is completely dissolved in distilled water according to 1: 25 mass ratio, adds Sevage reagents by cumulative volume 1/4, fully vibrates 2h, 3500r/min centrifuges 10min, and material points three layers, intermediate layer is solid material, collects supernatant liquor, then to intermediate layer and The material of lower floor repeats the step and handled untill intermediate layer is without solid-like floating preteins, with Rotary Evaporators on above-mentioned Layer clear liquid is concentrated to give concentrate, 4 times of volume absolute ethyl alcohols is added in the concentrate, 4 DEG C stand overnight, then 3500r/min Centrifugation 5min takes precipitation.
- 6. extracting method as claimed in claim 1, it is characterised in that：The step (5) is：With distilled water dissolving step (4) The precipitation of gained, concentrated with Rotary Evaporators after multigelation, use molecular cut off to be dialysed for 3500 bag filter flowing water 24h, distilled water dialysis 24h, freeze-drying obtain the second crude extract, and the condition of the multigelation of the step is：Cryogenic temperature for- 20 DEG C, solution temperature is 50 DEG C, is repeated 5 times.
- 7. extracting method as claimed in claim 1, it is characterised in that：The bag filter used in dialysis in the step (6) Molecular cut off is 3300~3800.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711094188.0A CN107868136B (en) | 2017-11-08 | 2017-11-08 | Method for extracting polysaccharide from herba Bulbophylli Inconspicui with anticancer effect |
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| CN201711094188.0A CN107868136B (en) | 2017-11-08 | 2017-11-08 | Method for extracting polysaccharide from herba Bulbophylli Inconspicui with anticancer effect |
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| CN107868136B CN107868136B (en) | 2020-03-10 |
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Cited By (2)
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| CN113278090A (en) * | 2021-06-28 | 2021-08-20 | 华侨大学 | Albizzia julibrissin polysaccharide, preparation method and application |
| CN113667030A (en) * | 2021-08-11 | 2021-11-19 | 华南理工大学 | Method for decoloring and purifying osmanthus fragrans crude polysaccharide |
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| CN106632716A (en) * | 2016-12-23 | 2017-05-10 | 桂林医学院 | Application of pholidota chinensis lindl. polysaccharides to preparation of hepatoprotective |
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| HAIHUA YANG等: "Characterization and antioxidant activity of a novel polysaccharidefrom Pholidota chinensis Lindl.", 《CARBOHYDRATE POLYMERS》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113278090A (en) * | 2021-06-28 | 2021-08-20 | 华侨大学 | Albizzia julibrissin polysaccharide, preparation method and application |
| CN113667030A (en) * | 2021-08-11 | 2021-11-19 | 华南理工大学 | Method for decoloring and purifying osmanthus fragrans crude polysaccharide |
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