CN104292353B - A kind of antioxidation natural plant polyose and preparation method thereof - Google Patents
A kind of antioxidation natural plant polyose and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the field of Chinese medicines, be specifically related to a kind of antioxidation natural plant polyose.The present invention is with natural Chinese medicinal herb Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae and Herba Epimedii for core, its active ingredient Polyose extraction has been extracted, and the formula of maximum oxidation resistance is obtained through scientific and reasonable compatibility, the formula invented serves the potentiation that three taste source medicine polysaccharide complementations are collaborative.Polysaccharide component in formula derives from natural plants, without chemosynthesis composition, without hormone, heavy metal or any western medicine composition, has the pharmacodynamic features such as stronger antioxidation, defying age and immunity lifting, great industrialization development potentiality.
Description
Technical field
The invention belongs to the field of Chinese medicines, be specifically related to a kind of antioxidation natural plant polyose.
Background technology
In recent years, along with society, life, work, environment change, human diseases spectrum there occurs huge change.The disease that incidence rate is high and harm is maximum has been turned to new hair style variability virus, body aging or body own metabolism by traditional infectious single factor test disease and has regulated and controled not normal for the main complexity multifactor property disease composing group.Under this trend, numerous be target with single disease target spot conventional medicament increasingly can not meet the needs of mankind's preventing and treating diseases.Therefore, from enhancing body non-oxidizability and autoimmunity, utilize the distinctive multicomponent of Chinese herb medicine, too many levels, Mutiple Targets Integrate adjustment ability flight against senium of human body antioxidation and immune system are improved, thus reaching to improve the purpose of human health level, it has also become medical treatment or a focus in health care activity.
Modern medicine study achievement shows, a lot of Chinese herbal medicine contain polysaccharide, organic acid, alkaloids, glycoside and volatile oil etc., these materials pass through to promote the oxidation resistance of body in human body, strengthen immune cell function, improve the effects such as cytokine secretion, the consolidation to each organ dysfunction of body and lifting can be realized, the preventing and treating of disease is had great significance.
Herba Scutellariae Barbatae, Herba Hedyotidis Diffusae and Herba Epimedii are the primary medicinal plants of Funiu Mountains of Henan Province In China.Recording according to tradition medical book, this three herbal medicine has invigorating the spleen and replenishing QI, blood circulation promoting and blood stasis dispelling, and effect of heat-clearing and toxic substances removing, as reasonable compatibility uses, when playing an important role in antioxidation and immunity lifting.By modern science bibliographical information, Scutellaria Barbata D. Don Polysaccharides can promote the ConA mouse boosting cell lymphocyte transformation induced in vitro, and its optimum concentration is 400 μ g/ml.Subcutaneous administrations one Zhou Houke significantly improves the percentage rate of esterase positive cell in Mouse Peripheral Blood Lymphocyte, promotes the DNCB mice delayed allergy brought out after intraperitoneal administration.Herba Scutellariae Barbatae water carries polysaccharide to H2O2There is stronger scavenging action, and in dose-effect relationship.Showing through mouse experiment, its oxidation-resisting and caducity effect is to resist old and feeble life-saving by raising SOD vigor and scavenging action and lipoid peroxidization resistant to negative oxygen-derived free radicals.Herba Hedyotidis Diffusae polysaccharide extract can strengthen immunity phagocyte activity, the proliferation activity of mouse boosting cell there is very strong facilitation, the generation and the monocytic cytokine that strengthen B cell antibody produce, strengthen the mononuclear cell phagocytic function to tumor cell, and promote the secretion of proliferation of bone marrow cells reaction and IL2.It addition, Herba Hedyotidis Diffusae polysaccharide has the effect of enhancing body specific immune function and non-specific immunity.Herba Epimedii can improve the phagocytic function of Turnover of Mouse Peritoneal Macrophages and the phagocytic function being subject to the Turnover of Mouse Peritoneal Macrophages of cyclophosphamide damage can be made to recover to normal level.The phagocytic function of immunologic hypofunction model mice macrophage caused by heavy dose of hydrocortisone and hydroxyurea is remarkably reinforced effect by Herba Epimedii.Macrophages secrete IL-1 and tumor necrosis factor are had dual regulation by epimedium brevicornum polysaccharide, and promote the lymphocytic proliferation and differentiation of T/B and natural killer cell activity, improve Immune Function.It addition, Herba Epimedii can promote the secretion of panimmunity cytokine, such as interferon and IL-2 etc..
In sum, tradition medicinal plants Herba Scutellariae Barbatae, no matter Herba Hedyotidis Diffusae and Herba Epimedii are based on Traditional Chinese medical theory, or modern pharmacology Research foundation, all show extremely strong antioxidant activity and the potentiality of enhancing human body immunity function, its chief active position is polysaccharide.Therefore, targeting optimization separates Herba Scutellariae Barbatae, Herba Hedyotidis Diffusae and epimedium brevicornum polysaccharide, and carries out composite under drug effect instructs, and can be achieved to the compound prescription of a kind of effective antioxidation natural plant polyose.
Summary of the invention
It is an object of the invention to provide a kind of antioxidation natural plant polyose, under the premise without any synthetic chemical composition, exploitation has antioxidation, anti-aging and promote the natural plant polyose of immunity.
The present invention solves the problems referred to above, taked technical scheme be: a kind of antioxidation natural plant polyose, described vegetable polysaccharides is made up of Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide, and the weight ratio of each component is 1:1:1 needed for preparing vegetable polysaccharides；
Described Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are that respectively through ultrasonic Treatment, Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae and Herba Epimedii are prepared Herba Hedyotidis Diffusae extracting solution, Herba Scutellariae Barbatae extracting solution and extraction ofHerba Epimedii, then are evaporated prepared further by Herba Hedyotidis Diffusae extracting solution, Herba Scutellariae Barbatae extracting solution and extraction ofHerba Epimedii respectively.
The preparation method of described antioxidation natural plant polyose, comprises the following steps:
Step one, Herba Hedyotidis Diffusae polysaccharide preparation
Herba Hedyotidis Diffusae after pulverizing and distilled water are pressed the weight ratio mixing of 1:14, it is the ultrasonic extraction 30-40min of 40kHz by frequency afterwards, extracting solution aperture is the filter paper filtering of 20 μm, collect filtrate, repeat said extracted process 2-3 time, merging filtrate, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture I, the ethanol that volumetric concentration is 95% is added to mixture I, after making mixing, the volumetric concentration of ethanol is 60%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely Herba Hedyotidis Diffusae polysaccharide is prepared after filtrate being evaporated；
Step 2, Scutellaria Barbata D. Don Polysaccharides preparation
Herba Scutellariae Barbatae after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Scutellariae Barbatae is corresponding is 14ml, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat 5-6 rear merging filtrate of said extracted process, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture II, the ethanol that concentration of volume percent is 95% is added to mixture II, after making mixing, the volumetric concentration of ethanol is 80%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely Scutellaria Barbata D. Don Polysaccharides is prepared after filtrate being evaporated；
Step 3, epimedium brevicornum polysaccharide preparation
Herba Epimedii after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Epimedii is corresponding is 10ml, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat 4-5 rear merging filtrate of said extracted process, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture III, the ethanol that volumetric concentration is 95% is added to mixture III, after making mixing, the volumetric concentration of ethanol is 70%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely epimedium brevicornum polysaccharide is prepared after filtrate being evaporated；
Step 4, preparation
Prepared Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are mixed and stirred for after uniformly namely prepare vegetable polysaccharides by the weight ratio of 1:1:1.
Beneficial effect
The present invention is with natural Chinese medicinal herb Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae and Herba Epimedii for core, its active ingredient Polyose extraction has been extracted, and the formula of maximum oxidation resistance is obtained through scientific and reasonable compatibility, the formula invented serves the potentiation that three taste source medicine polysaccharide complementations are collaborative.Polysaccharide component in formula derives from natural plants, without chemosynthesis composition, without hormone, heavy metal or any western medicine composition, has the pharmacodynamic features such as stronger antioxidation, defying age and immunity lifting, great industrialization development potentiality.
Detailed description of the invention
A kind of antioxidation natural plant polyose, described vegetable polysaccharides is made up of Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide, and the weight ratio of each component is 1:1:1 needed for preparing vegetable polysaccharides；
Described Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are that respectively through ultrasonic Treatment, Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae and Herba Epimedii are prepared Herba Hedyotidis Diffusae extracting solution, Herba Scutellariae Barbatae extracting solution and extraction ofHerba Epimedii, then are evaporated prepared further by Herba Hedyotidis Diffusae extracting solution, Herba Scutellariae Barbatae extracting solution and extraction ofHerba Epimedii respectively.
The preparation method of described antioxidation natural plant polyose, comprises the following steps:
Step one, Herba Hedyotidis Diffusae polysaccharide preparation
Herba Hedyotidis Diffusae after pulverizing and distilled water are pressed the weight ratio mixing of 1:14, it is the ultrasonic extraction 30-40min of 40kHz by frequency afterwards, extracting solution aperture is the filter paper filtering of 20 μm, collect filtrate, repeat said extracted process 2-3 time, merging filtrate, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture I, the ethanol that volumetric concentration is 95% is added to mixture I, after making mixing, the volumetric concentration of ethanol is 60%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely Herba Hedyotidis Diffusae polysaccharide is prepared after filtrate being evaporated；
Step 2, Scutellaria Barbata D. Don Polysaccharides preparation
Herba Scutellariae Barbatae after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Scutellariae Barbatae is corresponding is 14ml, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat 5-6 rear merging filtrate of said extracted process, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture II, the ethanol that concentration of volume percent is 95% is added to mixture II, after making mixing, the volumetric concentration of ethanol is 80%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely Scutellaria Barbata D. Don Polysaccharides is prepared after filtrate being evaporated；
Step 3, epimedium brevicornum polysaccharide preparation
Herba Epimedii after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Epimedii is corresponding is 10ml, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat 4-5 rear merging filtrate of said extracted process, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture III, the ethanol that volumetric concentration is 95% is added to mixture III, after making mixing, the volumetric concentration of ethanol is 70%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely epimedium brevicornum polysaccharide is prepared after filtrate being evaporated；
Step 4, preparation
Prepared Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are mixed and stirred for after uniformly namely prepare vegetable polysaccharides by the weight ratio of 1:1:1.
Embodiment 1
A kind of antioxidation natural plant polyose, described vegetable polysaccharides is made up of Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide, and the weight ratio of each component is 1:1:1 needed for preparing vegetable polysaccharides；
Described Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are that respectively through ultrasonic Treatment, Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae and Herba Epimedii are prepared Herba Hedyotidis Diffusae extracting solution, Herba Scutellariae Barbatae extracting solution and extraction ofHerba Epimedii, then are evaporated prepared further by Herba Hedyotidis Diffusae extracting solution, Herba Scutellariae Barbatae extracting solution and extraction ofHerba Epimedii respectively.
The preparation method of described antioxidation natural plant polyose, comprises the following steps:
Step one, Herba Hedyotidis Diffusae polysaccharide preparation
Herba Hedyotidis Diffusae after pulverizing and distilled water are pressed the weight ratio mixing of 1:14, it is the ultrasonic extraction 30min of 40kHz by frequency afterwards, extracting solution aperture is the filter paper filtering of 20 μm, collect filtrate, repeat said extracted process 2 times, merging filtrate, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture I, adding, to mixture I, the ethanol that volumetric concentration is 95%, after making mixing, the volumetric concentration of ethanol is 60%, after standing 24h, with the filter paper filtering that aperture is 20 μm, after filtrate being evaporated, namely prepare Herba Hedyotidis Diffusae polysaccharide；
Step 2, Scutellaria Barbata D. Don Polysaccharides preparation
Herba Scutellariae Barbatae after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Scutellariae Barbatae is corresponding is 14ml, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat merging filtrate after said extracted process 5 times, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture II, the ethanol that concentration of volume percent is 95% is added to mixture II, after making mixing, the volumetric concentration of ethanol is 80%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely Scutellaria Barbata D. Don Polysaccharides is prepared after filtrate being evaporated；
Step 3, epimedium brevicornum polysaccharide preparation
Herba Epimedii after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Epimedii is corresponding is 10ml, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat 4-5 rear merging filtrate of said extracted process, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture III, the ethanol that volumetric concentration is 95% is added to mixture III, after making mixing, the volumetric concentration of ethanol is 70%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely epimedium brevicornum polysaccharide is prepared after filtrate being evaporated；
Step 4, preparation
Prepared Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are mixed and stirred for after uniformly namely prepare vegetable polysaccharides by the weight ratio of 1:1:1.
Embodiment 2
A kind of antioxidation natural plant polyose, described vegetable polysaccharides is made up of Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide, and the weight ratio of each component is 1:1:1 needed for preparing vegetable polysaccharides；
Described Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are that respectively through ultrasonic Treatment, Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae and Herba Epimedii are prepared Herba Hedyotidis Diffusae extracting solution, Herba Scutellariae Barbatae extracting solution and extraction ofHerba Epimedii, then are evaporated prepared further by Herba Hedyotidis Diffusae extracting solution, Herba Scutellariae Barbatae extracting solution and extraction ofHerba Epimedii respectively.
The preparation method of described antioxidation natural plant polyose, comprises the following steps:
Step one, Herba Hedyotidis Diffusae polysaccharide preparation
Herba Hedyotidis Diffusae after pulverizing and distilled water are pressed the weight ratio mixing of 1:14, it is the ultrasonic extraction 40min of 40kHz by frequency afterwards, extracting solution aperture is the filter paper filtering of 20 μm, collect filtrate, repeat said extracted process 3 times, merging filtrate, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture I, adding, to mixture I, the ethanol that volumetric concentration is 95%, after making mixing, the volumetric concentration of ethanol is 60%, after standing 24h, with the filter paper filtering that aperture is 20 μm, after filtrate being evaporated, namely prepare Herba Hedyotidis Diffusae polysaccharide；
Step 2, Scutellaria Barbata D. Don Polysaccharides preparation
Herba Scutellariae Barbatae after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Scutellariae Barbatae is corresponding is 14ml, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat merging filtrate after said extracted process 6 times, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture II, the ethanol that concentration of volume percent is 95% is added to mixture II, after making mixing, the volumetric concentration of ethanol is 80%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely Scutellaria Barbata D. Don Polysaccharides is prepared after filtrate being evaporated；
Step 3, epimedium brevicornum polysaccharide preparation
Herba Epimedii after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Epimedii is corresponding is 10ml, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat 4-5 rear merging filtrate of said extracted process, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture III, the ethanol that volumetric concentration is 95% is added to mixture III, after making mixing, the volumetric concentration of ethanol is 70%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely epimedium brevicornum polysaccharide is prepared after filtrate being evaporated；
Step 4, preparation
Prepared Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are mixed and stirred for after uniformly namely prepare vegetable polysaccharides by the weight ratio of 1:1:1.
Experiment one, the collocation of whole formula and preparation can be divided into following four step
One, Herba Hedyotidis Diffusae polysaccharide preparation technology
It is as follows that Herba Hedyotidis Diffusae polysaccharide extracts preparation process, Herba Hedyotidis Diffusae is ground into powder, mixes with certain solid-liquid ratio with distilled water, carries out supersound extraction with the time set, and crosses leaching supernatant, repeats to extract once, merges supernatant.Rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach preset percentage, stands overnight, be filtered to remove insoluble matter, be evaporated to obtain polysaccharide.For optimizing polysaccharide yield and purity, first carry out experiment of single factor as follows:
1, Herba Hedyotidis Diffusae polysaccharide extracts experiment of single factor
A) different feed liquid is than test
Set solid-liquid ratio (g/ml):
Material: Spreading Hedyotis Herb 5g, distilled water
1:6,1:10,1:14,1:18,1:24.
Putting into ultrasonic Treatment 30min, cross leaching supernatant, repeat to extract once, merge supernatant, rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, and anthrone colorimetry measures polysaccharide.
B) test
Solid-liquid ratio 1:10(g/ml)
Material: Spreading Hedyotis Herb 5g, distilled water
Set and put into ultrasonic time 10min, 20min, 30min, 40min, 50min.
Crossing leaching supernatant, repeat to extract once, merge supernatant, rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, and anthrone colorimetry measures polysaccharide.
C) different extraction times
Solid-liquid ratio 1:10(g/ml)
Material: Spreading Hedyotis Herb 5g, distilled water
Ultrasonic 30min is put in setting, crosses leaching supernatant, sets and repeats extraction 1,2,3,4,5, secondary, merge supernatant, rotary evaporation is to original volume 1/4th, adding ethanol, make concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, anthrone colorimetry measures polysaccharide.
D)
Solid-liquid ratio 1:10(g/ml)
Material: Spreading Hedyotis Herb 5g, distilled water
Setting and put into ultrasonic time 30min, cross leaching supernatant, repeat to extract once, merge supernatant, rotary evaporation is to original volume 1/4th, add ethanol, make concentration of alcohol reach 50%, 60%, 70%, 80%, 90%, overnight it is filtered to remove insoluble matter afterwards, is evaporated to obtain polysaccharide, anthrone colorimetry measures polysaccharide.
In experiment of single factor, each obtains the light absorption value (i.e. polyoses content) of sample
Thereby determining that, in orthogonal test, parameter solid-liquid ratio is 1:6,1:10 and 1:14；Ultrasonic time is 20min, 30min and 40min；Extraction time is 2,3 and 4 times；Concentration of alcohol is 60%, 70% and 90%.
2, Herba Hedyotidis Diffusae polysaccharide extracts orthogonal test
Orthogonal test parameter
Parameter designed in table is used to proceed as follows:
Solid-liquid ratio is the ratio of the quality of Spreading Hedyotis Herb and distilled water, and the quality of every part of Herba Hedyotidis Diffusae is 5g；The solution (quality of every part of medical material is 5g) prepared in proportion is put in ultrasound wave；The ultrasonic time reaching defined is stop 1 minute in every 5 minutes；Cross leaching supernatant, repeat to extract, after conjunction and supernatant；Rotary evaporation, to original volume 1/4th, adds the ethanol content of regulation, overnight；It is filtered to remove insoluble matter, is evaporated to obtain polysaccharide；Anthrone colorimetry measures polysaccharide.
Orthogonal experiments is analyzed
It follows that the optimum extraction parameter of Herba Hedyotidis Diffusae polysaccharide is: solid-liquid ratio=1:14；Extraction time 30-40min；Repeat extraction time 3 times；Extract concentration of alcohol 60%.
Two, Scutellaria Barbata D. Don Polysaccharides preparation technology
It is as follows that Scutellaria Barbata D. Don Polysaccharides extracts preparation process, Herba Scutellariae Barbatae is ground into powder, mixes with certain solid-liquid ratio with distilled water, carries out supersound extraction with the time set, and crosses leaching supernatant, repeats to extract once, merges supernatant.Rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach preset percentage, stands overnight, be filtered to remove insoluble matter, be evaporated to obtain polysaccharide.For optimizing polysaccharide yield and purity, first carry out experiment of single factor as follows:
1, Scutellaria Barbata D. Don Polysaccharides extracts experiment of single factor
A) different feed liquid is than test
Material: Herba Scutellariae Barbatae powder 5g, distilled water
Set solid-liquid ratio (g/ml): 1:6,1:10,1:14,1:18,1:24.
Putting into ultrasonic time 30min, cross leaching supernatant, repeat to extract once, merge supernatant, rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, and anthrone colorimetry measures polysaccharide.
B) different supersound extraction time tests
Solid-liquid ratio 1:10
Material: Herba Scutellariae Barbatae powder 5g, distilled water
Set and put into ultrasonic time 10min, 20min, 30min, 40min, 50min
Crossing leaching supernatant, repeat to extract once, merge supernatant, rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, and anthrone colorimetry measures polysaccharide.
C) different extraction times
Solid-liquid ratio 1:10
Material: Herba Scutellariae Barbatae powder 5g, distilled water
Ultrasonic Treatment 30min is put in setting, crosses leaching supernatant, sets and repeats extraction 1,2,3,4,5, secondary, merge supernatant, rotary evaporation is to original volume 1/4th, adding ethanol, make concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, anthrone colorimetry measures polysaccharide.
D) different ethanol concentration
Solid-liquid ratio 1:10
Material: Herba Scutellariae Barbatae powder 5g, distilled water
Setting and put into ultrasonic time 30min, cross leaching supernatant, repeat to extract once, merge supernatant, rotary evaporation is to original volume 1/4th, add ethanol, make concentration of alcohol reach 50%, 60%, 70%, 80%, 90%, overnight it is filtered to remove insoluble matter afterwards, is evaporated to obtain polysaccharide, anthrone colorimetry measures polysaccharide.
In experiment of single factor, each obtains the light absorption value (i.e. polyoses content) of sample
Thereby determining that, in orthogonal test, parameter solid-liquid ratio is 1:6,1:10 and 1:14；Ultrasonic time is 10min, 20min and 30min；Extraction time is 3,5 and 6 times；Concentration of alcohol is 60%, 80% and 90%.
2, Scutellaria Barbata D. Don Polysaccharides extracts orthogonal test
Orthogonal test parameter
Parameter designed in table is used to proceed as follows:
Solid-liquid ratio is distilled water on the mass ratio of Herba Scutellariae Barbatae powder, and the quality of every part of Herba Scutellariae Barbatae is 5g；The solution (quality of every part of medical material is 5g) prepared in proportion is put in ultrasound wave；The ultrasonic time reaching defined, every 5 minutes stop 1 minute；Cross leaching supernatant, repeat to extract, merge supernatant；Rotary evaporation, to original volume 1/4th, adds the ethanol content of regulation, overnight；It is filtered to remove insoluble matter, is evaporated to obtain polysaccharide；Anthrone colorimetry measures polysaccharide.
Orthogonal experiments is analyzed
It follows that the optimum extraction parameter of Scutellaria Barbata D. Don Polysaccharides is solid-liquid ratio=1:14；Extraction time=10min；Repeat extraction time=5 time；Extract concentration of alcohol=80%.
Three, epimedium brevicornum polysaccharide preparation technology
It is as follows that epimedium brevicornum polysaccharide extracts preparation process, Herba Epimedii is ground into powder, mixes with certain solid-liquid ratio with distilled water, carries out supersound extraction with the time set, and crosses leaching supernatant, repeats to extract once, merges supernatant.Rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach preset percentage, stands overnight, be filtered to remove insoluble matter, be evaporated to obtain polysaccharide.For optimizing polysaccharide yield and purity, first carry out experiment of single factor as follows:
1, epimedium brevicornum polysaccharide extracts experiment of single factor
A) different feed liquid is than test
Material: Epimedium Herb 5g, distilled water
Set solid-liquid ratio (g/ml) 1:6,1:10,1:14,1:18,1:24；
Put into ultrasonic time 30min；Cross leaching supernatant, repeat to extract once, merge supernatant；Rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, and anthrone colorimetry measures polysaccharide.
B) different supersound extraction time tests
Solid-liquid ratio 1:10
Material: Epimedium Herb 5g, distilled water
Set and put into ultrasonic time 10min, 20min, 30min, 40min, 50min
Crossing leaching supernatant, repeat to extract once, merge supernatant, rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, and anthrone colorimetry measures polysaccharide.
C) different extraction times
Solid-liquid ratio 1:10
Material: Epimedium Herb 5g, distilled water
Ultrasonic 30min is put in setting；Crossing leaching supernatant, set and repeat extraction 1,2,3,4,5 time, merge supernatant, rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach 60%, be overnight filtered to remove insoluble matter afterwards, be evaporated to obtain polysaccharide, and anthrone colorimetry measures polysaccharide.
D) different ethanol concentration
Solid-liquid ratio 1:10
Material: Epimedium Herb 5g, distilled water
Set and put into ultrasonic time 30min；Cross leaching supernatant, repeat to extract once, merge supernatant；Rotary evaporation, to original volume 1/4th, adds ethanol, makes concentration of alcohol reach 50%, and 60%, 70%, 80%, 90%, overnight it is filtered to remove insoluble matter afterwards, is evaporated to obtain polysaccharide, anthrone colorimetry measures polysaccharide.
In experiment of single factor, each obtains the light absorption value (i.e. polyoses content) of sample
Thereby determining that, in orthogonal test, parameter solid-liquid ratio is 1:6,1:10 and 1:22；Ultrasonic time is 10,30 and 50min；Extraction time is 2,5 and 6 times；Concentration of alcohol is 70%, 80% and 90%.
2, epimedium brevicornum polysaccharide extracts orthogonal test
Orthogonal test parameter
Parameter designed in table is used to proceed as follows:
Solid-liquid ratio is distilled water on the mass ratio of Epimedium Herb, and the quality of every part of Herba Epimedii is 5g；The solution (quality of every part of medical material is 5g) prepared in proportion is put in ultrasound wave；Ultrasound wave to reach the time of defined, and every 5 minutes stop 1 minute；Cross leaching supernatant, repeat to extract, after conjunction and supernatant；Rotary evaporation, to original volume 1/4th, adds the ethanol content of regulation, overnight；It is filtered to remove insoluble matter, is evaporated to obtain polysaccharide；Anthrone colorimetry measures polysaccharide.
Orthogonal experiments is analyzed
It follows that the optimum extraction parameter of epimedium brevicornum polysaccharide is: solid-liquid ratio=1:10；Extraction time=10min；Repeat extraction time=4 time；Extract concentration of alcohol=70%.
Four, Herba Hedyotidis Diffusae-Herba Scutellariae Barbatae-Herba Epimedii polysaccharide formula and preparation
For determining Herba Hedyotidis Diffusae, the optimum proportioning of Herba Scutellariae Barbatae and epimedium brevicornum polysaccharide, nine groups of proportionings of ad hoc meter, by three kinds of polysaccharide respectively with Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=1:1:1；Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=2:1:2；Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=3:1:3；Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=1:2:2；Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=2:2:3；Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=3:2:1；Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=1:3:3；Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=2:3:1；Herba Hedyotidis Diffusae: Herba Scutellariae Barbatae: Herba Epimedii=3:3:2 mixes.
To nine groups of samples DPPH free radical scavenging activity being measured under variable concentrations obtained, thus obtaining its oxidation resistance, result is following table such as:
In table in vain: half: excessive, Herba Hedyotidis Diffusae polysaccharide is represented: Scutellaria Barbata D. Don Polysaccharides: epimedium brevicornum polysaccharide.
Using orthogonal test that the proportioning parameters relationship of Herba Hedyotidis Diffusae in formula, Herba Scutellariae Barbatae and epimedium brevicornum polysaccharide is analyzed, its result is as follows:
Orthogonal experiments is analyzed
By result above it may be concluded that when Herba Hedyotidis Diffusae polysaccharide: Scutellaria Barbata D. Don Polysaccharides: when epimedium brevicornum polysaccharide=1:1:1 carries out compatibility, formula can obtain the antioxidant activity of the best.
Claims (1)
1. an antioxidation natural plant polyose, it is characterised in that: its preparation method comprises the following steps:
Step one, Herba Hedyotidis Diffusae polysaccharide preparation
Herba Hedyotidis Diffusae after pulverizing and distilled water are pressed the weight ratio mixing of 1:14, it is the ultrasonic extraction 30-40min of 40kHz by frequency afterwards, extracting solution aperture is the filter paper filtering of 20 μm, collect filtrate, repeat said extracted process 2-3 time, merging filtrate, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture I, the ethanol that volumetric concentration is 95% is added to mixture I, after making mixing, the volumetric concentration of ethanol is 60%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely Herba Hedyotidis Diffusae polysaccharide is prepared after filtrate being evaporated；
Step 2, Scutellaria Barbata D. Don Polysaccharides preparation
Herba Scutellariae Barbatae after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Scutellariae Barbatae is corresponding is 14mL, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat 5-6 rear merging filtrate of said extracted process, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture II, the ethanol that concentration of volume percent is 95% is added to mixture II, after making mixing, the volumetric concentration of ethanol is 80%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely Scutellaria Barbata D. Don Polysaccharides is prepared after filtrate being evaporated；
Step 3, epimedium brevicornum polysaccharide preparation
Herba Epimedii after pulverizing and distilled water are mixed and stirred for uniformly, the distilled water volume that every gram of Herba Epimedii is corresponding is 10mL, it is the ultrasonic extraction 10min of 40kHz by frequency afterwards, by the filter paper filtering that extracting solution aperture is 20 μm, collect filtrate, repeat 4-5 rear merging filtrate of said extracted process, and filtrate is put in Rotary Evaporators and be concentrated into original volume 1/4th, prepare mixture III, the ethanol that volumetric concentration is 95% is added to mixture III, after making mixing, the volumetric concentration of ethanol is 70%, after standing 24h, with the filter paper filtering that aperture is 20 μm, namely epimedium brevicornum polysaccharide is prepared after filtrate being evaporated；
Step 4, preparation
Prepared Herba Hedyotidis Diffusae polysaccharide, Scutellaria Barbata D. Don Polysaccharides and epimedium brevicornum polysaccharide are mixed and stirred for after uniformly namely prepare vegetable polysaccharides by the weight ratio of 1:1:1.
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| CN107216401A (en) * | 2016-12-23 | 2017-09-29 | 翁绍瑀 | A kind of preparation method of Epimedium polysaccharide |
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