CN105254777A - Extraction method of Yupingfeng polysaccharide - Google Patents
Extraction method of Yupingfeng polysaccharide Download PDFInfo
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Abstract
The invention discloses an extraction method of Yupingfeng polysaccharide. The method comprises the steps that 1, milkvetch roots, saposhnicovia divaricata and largehead atractylodes rhizome are prepared into Yupingfeng powder; 2, the Yupingfeng powder is put into deionized water to be soaked for 20-40 min and then put into a water bath to conduct reflux extraction, and extraction is conducted for three times; 3, cooling is conducted after extraction is conducted, filtration is conducted through four layers of gauze, filtrate is collected, vacuum filtration is conducted for two times, the filtrate obtained through vacuum filtration is collected to be heated and concentrated through warm fire, and a concentrated solution is cooled to room temperature; 4, absolute ethyl alcohol of which the volume is four times of that of the concentrated solution is added to be fully stirred, standing is conducted for a period of time at the room temperature, upper layer mixed liquor is poured out, and precipitate is taken out; 5, the precipitate is frozen and put in the vacuum environment to be frozen and dried, and the powdered Yupingfeng polysaccharide can be obtained.
Description
Technical field
The present invention relates to field of biological pharmacy, particularly relate to a kind of extracting method of Chinese medicine Yupingfeng polysaccharide.
Background technology
Yupingfeng San, comes from unit. and Wei Yilin Effective Formulas Tested by Physicians for Generations, has benefiting QI for strengthening the superficies, righting, eliminating evil effect.In side, the Radix Astragali has effect of benefiting QI for strengthening the superficies hidroschesis, is monarch drug in a prescription; Bighead atractylodes rhizome invigorating the spleen tonifying Qi is ministerial drug; A left side makes medicine be windproof, walks table and wind dispelling cold of faling apart, and closes the Radix Astragali, the bighead atractylodes rhizome is eliminating evil with beneficial gas.Modern study shows, this can the immunologic function of enhancing body, and to weakened defensive QI, deficiency of vital energy table is weak, and the card of susceptible chill has good therapeutic action.
Yupingfeng polysaccharide is made up of astragalus polysaccharides, sposknikovan and soluble polysaccharide isoreactivity polysaccharide.Show according to correlative study, Yupingfeng polysaccharide can impel growing of the immune organs such as man and animal thymus gland, spleen, strengthens the activity of immunocyte, improves the immune effect of vaccine and postpones the time length of antibody, therefore, Yupingfeng polysaccharide has unique advantage in immunomodulatory.
Because Yupingfeng polysaccharide has great use value and marketable value as immunostimulant for animals, and the vacuum filtration desiccating method adopted in present stage Yupingfeng polysaccharide extracting process and method for filling are processing high purity, poor effect during viscosity large Yupingfeng polysaccharide.Highly purified Yupingfeng polysaccharide viscosity is very big, and the Yupingfeng polysaccharide pole be stored in a constant volume device is not easy to take out and weighs and come into operation.The characteristic that its viscosity is large makes Yupingfeng polysaccharide waste in a large number, so rate of loss is high because of sticky wall.
Therefore, prior art has yet to be improved and developed.
Summary of the invention
In view of above-mentioned the deficiencies in the prior art, the object of the present invention is to provide a kind of extracting method of Yupingfeng polysaccharide, it is not suitable for the problem of Yupingfeng polysaccharide to be intended to solve existing extracting method.
Technical scheme of the present invention is as follows:
An extracting method for Yupingfeng polysaccharide, wherein, comprises step:
A, the Radix Astragali, the windproof and bighead atractylodes rhizome are configured to Yupingfeng San;
B, Yupingfeng San is placed in deionized water and soaks 20 ~ 40min, then put refluxing extraction in water-bath, extract 3 times altogether;
C, extracted rear cooling, first filtered with 4 layers of gauze, collected filtrate and carry out vacuum filtration twice again, the filtrate of then collecting vacuum filtration gained is concentrated with the heating of temperature fire, and solution to be concentrated is cooled to room temperature;
D, add the dehydrated alcohol of concentrated solution volume four times, fully stir, room temperature leaves standstill for some time, is gone by upper strata mixed solution, taking precipitate;
E, to freeze freezing for throw out, then be positioned over lyophilize in vacuum environment, pulverous Yupingfeng Crude polysaccharides can be obtained.
The extracting method of described Yupingfeng polysaccharide, wherein, in described step B, after adding deionized water, the solid-liquid ratio of Yupingfeng San is 1:16.
The extracting method of described Yupingfeng polysaccharide, wherein, in described steps A, the Radix Astragali, mass ratio that the is windproof and bighead atractylodes rhizome are 3:1:1.
The extracting method of described Yupingfeng polysaccharide, wherein, in described step B, soak time is 30min.
The extracting method of described Yupingfeng polysaccharide, wherein, in described step B, refluxing extraction condition is: extract 1.5h in 90 DEG C.
The extracting method of described Yupingfeng polysaccharide, wherein, in described step D, room temperature leaves standstill 12 hours.
The extracting method of described Yupingfeng polysaccharide, wherein, in described step e, lyophilize 12 hours in vacuum environment.
Beneficial effect: by the improvement of producing Yupingfeng polysaccharide process, the present invention can produce pulverous Yupingfeng polysaccharide in a large number, improves the sticky wall problem that Yupingfeng polysaccharide in the past occurs because purity is high, viscosity large.
Accompanying drawing explanation
Fig. 1 is Glucose standards solution curve in the present invention.
Embodiment
The invention provides a kind of extracting method of Yupingfeng polysaccharide, for making object of the present invention, technical scheme and effect clearly, clearly, the present invention is described in more detail below.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
The extracting method of a kind of Yupingfeng polysaccharide provided by the present invention, wherein, comprises step:
S1, the Radix Astragali, the windproof and bighead atractylodes rhizome are configured to Yupingfeng San;
S2, Yupingfeng San is placed in deionized water and soaks 20 ~ 40min, then put refluxing extraction in water-bath, extract 3 times altogether;
S3, extracted rear cooling, first filtered with 4 layers of gauze, collected filtrate and carry out vacuum filtration twice again, the filtrate of then collecting vacuum filtration gained is concentrated with the heating of temperature fire, and solution to be concentrated is cooled to room temperature;
S4, add the dehydrated alcohol of concentrated solution volume four times, fully stir, room temperature leaves standstill for some time, is gone by upper strata mixed solution, taking precipitate;
S5, to freeze freezing for throw out, then be positioned over lyophilize in vacuum environment, pulverous Yupingfeng Crude polysaccharides can be obtained.
In step sl, each raw material can be made medicinal powder, then by medicinal powder in mass ratio 3:1:1 be mixed with the Yupingfeng San of Radix Astragali 18g, windproof 6g and bighead atractylodes rhizome 6g altogether 30g, be placed in the round-bottomed flask of 1L.
Further, in described step S2, add deionized water in round-bottomed flask after, last solid-liquid ratio is 1:16(w:v).Then soak 20 ~ 40min(and be preferably 30min), put refluxing extraction in water-bath (extracting 1.5h in 90 DEG C), extract 3 times altogether.
In described step S3, be cooled to room temperature after having extracted, first filter with 4 layers of gauze, get filtrate and carry out vacuum filtration (two layers of filter paper) twice again, get filtrate and concentrate with temperature fire, solution to be concentrated is cooled to room temperature.
In step S4, room temperature time of repose is preferably 12 hours.In step S5, lyophilize 12 hours in vacuum environment.
Embodiment 1
(1) by medicinal powder in mass ratio 3:1:1 be mixed with the Yupingfeng San of Radix Astragali 18g, the windproof and each 6g of bighead atractylodes rhizome 30g altogether, be placed in the round-bottomed flask of 1L;
(2) Yupingfeng San is placed in respective amount deionized water by solid-liquid ratio (w:v) (1:16) and soaks 30min, put refluxing extraction in water-bath (extracting 1.5h in 90 DEG C), extract 3 times;
(3) extracted rear cooling, first filtered with 4 layers of gauze, got filtrate and carry out vacuum filtration (two layers of filter paper) twice again, get filtrate, with temperature fire, extracting solution is concentrated into 60ml, solution to be concentrated is cooled to room temperature;
(4) add the dehydrated alcohol that tetraploid is long-pending, stir, dehydrated alcohol is fully contacted with the effective constituent of polysaccharide (as stirred 15min) with glass stick, then room temperature leaves standstill 12 hours, is gone by upper strata mixed solution, taking precipitate (recovery ethanol);
(5) throw out is put into that refrigerator-20 DEG C of layers are freezing to freeze, then through vacuum freeze drier be in the vacuum environment of-50 DEG C dry 12 hours in temperature, be i.e. available pulverous Yupingfeng Crude polysaccharides.To be dispensed in cell therefor and to put Refrigerator store after sealing stand-by.
Embodiment 2
(1) by medicinal powder in mass ratio 4:1:1 be mixed with the Yupingfeng San of Radix Astragali 24g, the windproof and each 6g of bighead atractylodes rhizome 36g altogether, be placed in the round-bottomed flask of 1L;
(2) Yupingfeng San is placed in respective amount deionized water by solid-liquid ratio (w:v) (1:12) and soaks 20min, put refluxing extraction in water-bath (extracting 1h in 92 DEG C), extract 4 times;
(3) extracted rear cooling, first filtered with 4 layers of gauze, got filtrate and carry out vacuum filtration (two layers of filter paper) twice again, get filtrate, with temperature fire, extracting solution is concentrated into 60ml, solution to be concentrated is cooled to room temperature;
(4) add the dehydrated alcohol that tetraploid is long-pending, stir, dehydrated alcohol is fully contacted with the effective constituent of polysaccharide (as stirred 10min) with glass stick, then room temperature leaves standstill 12 hours, is gone by upper strata mixed solution, taking precipitate (recovery ethanol);
(5) throw out is put into that refrigerator-20 DEG C of layers are freezing to freeze, then through vacuum freeze drier be in the vacuum environment of-50 DEG C dry 12 hours in temperature, be i.e. available pulverous Yupingfeng Crude polysaccharides.To be dispensed in cell therefor and to put Refrigerator store after sealing stand-by.
Embodiment 3
(1) by medicinal powder in mass ratio 2:1:1 be mixed with the Yupingfeng San of Radix Astragali 12g, the windproof and each 6g of bighead atractylodes rhizome 24g altogether, be placed in the round-bottomed flask of 1L;
(2) Yupingfeng San is placed in respective amount deionized water by solid-liquid ratio (w:v1:18) and soaks 40min, put refluxing extraction in water-bath (extracting 2h in 93 DEG C), extract 2 times;
(3) extracted rear cooling, first filtered with 4 layers of gauze, got filtrate and carry out vacuum filtration (two layers of filter paper) twice again, get filtrate, with temperature fire, extracting solution is concentrated into 60ml, solution to be concentrated is cooled to room temperature;
(4) add the dehydrated alcohol that tetraploid is long-pending, stir, dehydrated alcohol is fully contacted with the effective constituent of polysaccharide (as stirred 20min) with glass stick, then room temperature leaves standstill 12 hours, is gone by upper strata mixed solution, taking precipitate (recovery ethanol);
(5) throw out is put into that refrigerator-20 DEG C of layers are freezing to freeze, then through vacuum freeze drier be in the vacuum environment of-50 DEG C dry 12 hours in temperature, be i.e. available pulverous Yupingfeng Crude polysaccharides.To be dispensed in cell therefor and to put Refrigerator store after sealing stand-by.
The extracting method that the present invention takes is the water extracting alcohol precipitator method of improvement.By to three kinds of Chinese medicines: the Radix Astragali, the bighead atractylodes rhizome, windproofly mixes, by a certain percentage at water-bath water-bath reflux to obtain its effective constituent.Therefore, extract the effective elements of the medicine in three kinds and need to control well the time and temperature of extracting.Consider the character of vegetable polysaccharides, the water temperature of water-bath is controlled to 90 ~ 93 DEG C by the present invention, and each water-bath return time is 1.5h, is divided into 3 times and extracts, so that the Effective Component of Chinese Medicine obtained is more, improves the productive rate of polysaccharide.
By electric furnace, polysaccharide extraction liquid is concentrated to a certain degree, when the near room temperature of temperature (a little more than 25 DEG C), pour dehydrated alcohol into wherein in the ratio of 1:4, in the process poured into, stir along a direction with glass stick, then by solution left standstill 12 hours.Pouring stirring during dehydrated alcohol into, in order to make the effective constituent of polysaccharide fully contact with dehydrated alcohol, being beneficial to the sedimentation of effective constituent.After concentrated solution temperature is down to certain degree, pours dehydrated alcohol into, is the contact in order to be beneficial to polysaccharide effective constituent and dehydrated alcohol equally, is beneficial to the sedimentation of polysaccharide effective constituent.
Show that the optimum process condition utilizing water extraction and alcohol precipitation method to extract Yupingfeng polysaccharide is by above-described embodiment and orthogonal test: solid-liquid ratio 1:16, Extracting temperature 90 DEG C, extraction time 1.5h, extraction time 3 times.The Yupingfeng polysaccharide that the present invention obtains is Powdered.Decrease the waste of polysaccharide to a certain extent, improve the output of polysaccharide; Pulverous Yupingfeng polysaccharide significantly improves the situation that polysaccharide in the past glues wall, is more convenient to preservation and the quantitative separating of Yupingfeng polysaccharide; Yupingfeng polysaccharide is mixed with certain density solution frozen, accurate quick more during use.
The extremely sticky wall of the Yupingfeng Crude polysaccharides obtained after alcohol precipitation, be difficult to scrape off, Crude polysaccharides is often bonded on wall because of can not scraping any more, and cause Crude polysaccharides to lose serious, extraction yield is had a greatly reduced quality.Same, if do not take process, directly polysaccharide freeze-drying in same container, obtain doing rule and loss being made even more serious and also can time consumption and energy consumption of Yupingfeng Crude polysaccharides.For this reason, the Crude polysaccharides of removing dehydrated alcohol is spread evenly across and speckles with on the culture dish of deionized water by the present invention, then is put into freeze-drying in vacuum freeze drier.Very thin one deck polysaccharide is uniformly coated on culture dish, increases the contact area of polysaccharide, make polysaccharide within a certain period of time freeze-drying obtain rapider, freeze-drying obtains more thorough, improves the efficiency of freeze-drying.Through multiple authentication, culture dish is sprayed an a small amount of water, polysaccharide is made to become puffy gradually during freeze-drying in vacuum freeze drier, the benefit done like this makes the polysaccharide of freeze-drying when scraping off, the polysaccharide be bonded on wall significantly reduces, solve polysaccharide and lose serious problem because of sticky wall, ensure that the productive rate of polysaccharide.Making the efficiency of scraping polysaccharide also increase substantially like this, avoiding when scraping Crude polysaccharides, Crude polysaccharides makes moist because of the impact of ambient moisture and becomes the problem of glutinous sticky wall.
Phenol-dense sulphuric acid method can be adopted in the present invention to measure the content of Yupingfeng polysaccharide:
1, the phenol Standard solution preparation of mass percent 5%
Take 5g phenol with beaker, add deionized water warm in right amount and dissolve, be transferred to 100mL volumetric flask after cooling, wash beaker 2-3 time with deionized water, washings all proceeds to volumetric flask, adds deionized water to scale, shakes up, carry out mark, is stored in 4 DEG C of refrigerators.
2, the preparation of Glucose standards solution
Accurately take the glucose 0.602g that 105 DEG C are dried to constant weight, be dissolved in 100mL deionized water, shake up dissolving, therefrom precision measures 10mL and is placed in 100mL volumetric flask, adds deionized water to scale, obtains the Glucose standards solution that concentration is 0.602mg/mL.
3, the drafting of typical curve
Precision measure Glucose standards solution 1.5,3.0,4.5,6.0,7.5,9.0mL is placed in 50mL volumetric flask, add deionized water and be settled to scale, precision measures 2mL to 25mL volumetric flask respectively, add the mixing of 1.0mL5% phenol solution more respectively, add rapidly the mixing of the 5.0mL vitriol oil, after leaving standstill 25min, under wavelength is 490nm, survey its absorbance, processing data drawing standard curve, take glucose concn as ordinate zou, absorbancy is that X-coordinate makes typical curve (see figure 1), obtains regression equation:
Y=75.83x-9.67，R
2=0.9946。
4, Yupingfeng determination of polysaccharide
Get respectively and prepare gained Yupingfeng polysaccharide 0.034g, 0.061g, 0.047g are placed in 100mL volumetric flask, are dissolved in water and are diluted to scale, shaking up, and therefrom precision measures 10mL and is placed in 100mL volumetric flask respectively, is dissolved in water and is diluted to scale, shake up.Measure its absorbance (mensuration wavelength chooses is 490nm) according to the method under Specification Curve of Increasing, calculate the content (first calculate polysaccharide concentration ug/mL, then calculate polysaccharide content) of polysaccharide.The results are shown in Table 1.
Table 1 Yupingfeng measurement of the polysaccharide content result
The acquisition of Yupingfeng polysaccharide final product still will by vacuum freeze drier, in addition will by complete for polysaccharide freeze-drying, also will control time of freeze-drying well.To the freeze-drying of Yupingfeng Crude polysaccharides, if freeze-drying time is not enough, then Crude polysaccharides is as easy as rolling off a log is bonded on wall, causes loss to a certain degree and product can stick into one, is unfavorable for packing.
Yupingfeng Crude polysaccharides viscosity before freeze-drying is very big, and extremely difficulty is scraped from container and taken out, and also storing its packing makes troubles.It is large equally to there is viscosity in the halfway Yupingfeng Crude polysaccharides of freeze-drying, the problem of difficult packing, and it is in block, is exposed in air, and extremely easily water suction is made moist.The present invention, by adjusting freeze-drying time (as freeze-drying 6 hours), can make pulverous polysaccharide, but its freeze-drying degree is still not enough, and it is still sticked on wall, not easily scraping during use.Further prolongation freeze-drying time (extending to 12 hours), the thorough freeze-drying of Crude polysaccharides, obtains pulverous Crude polysaccharides, the weak or not sticky wall of sticky wall degree.
Finally by obtained Yupingfeng Crude polysaccharides powder quantitative separating, conveniently can take.Further, Yupingfeng Crude polysaccharides powder is joined ultrapure water and dissolve, be mixed with certain density Yupingfeng polysaccharide soln, under low temperature, store (-20 DEG C), accurate quick more during use.
In sum, by the improvement of producing Yupingfeng polysaccharide process, the present invention can produce pulverous Yupingfeng polysaccharide in a large number, improves the sticky wall problem that Yupingfeng polysaccharide in the past occurs because purity is high, viscosity large.
Should be understood that, application of the present invention is not limited to above-mentioned citing, for those of ordinary skills, can be improved according to the above description or convert, and all these improve and convert the protection domain that all should belong to claims of the present invention.
Claims (7)
1. an extracting method for Yupingfeng polysaccharide, is characterized in that, comprises step:
A, the Radix Astragali, the windproof and bighead atractylodes rhizome are configured to Yupingfeng San;
B, Yupingfeng San is placed in deionized water and soaks 20 ~ 40min, then put refluxing extraction in water-bath, extract 3 times altogether;
C, extracted rear cooling, first filtered with 4 layers of gauze, collected filtrate and carry out vacuum filtration twice again, the filtrate of then collecting vacuum filtration gained is concentrated with the heating of temperature fire, and solution to be concentrated is cooled to room temperature;
D, add the dehydrated alcohol of concentrated solution volume four times, fully stir, room temperature leaves standstill for some time, is gone by upper strata mixed solution, taking precipitate;
E, to freeze freezing for throw out, then be positioned over lyophilize in vacuum environment, pulverous Yupingfeng Crude polysaccharides can be obtained.
2. the extracting method of Yupingfeng polysaccharide according to claim 1, is characterized in that, in described step B, after adding deionized water, the solid-liquid ratio of Yupingfeng San is 1:16.
3. the extracting method of Yupingfeng polysaccharide according to claim 1, is characterized in that, in described steps A, the Radix Astragali, mass ratio that the is windproof and bighead atractylodes rhizome are 3:1:1.
4. the extracting method of Yupingfeng polysaccharide according to claim 1, is characterized in that, in described step B, soak time is 30min.
5. the extracting method of Yupingfeng polysaccharide according to claim 1, is characterized in that, in described step B, refluxing extraction condition is: extract 1.5h in 90 DEG C.
6. the extracting method of Yupingfeng polysaccharide according to claim 1, is characterized in that, in described step D, room temperature leaves standstill 12 hours.
7. the extracting method of Yupingfeng polysaccharide according to claim 1, is characterized in that, in described step e, and lyophilize 12 hours in vacuum environment.
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Application publication date: 20160120 |