CN104744602A - Scrophularia ningpoensis polysaccharide and method for microwave extraction of scrophularia ningpoensis polysaccharide - Google Patents
Scrophularia ningpoensis polysaccharide and method for microwave extraction of scrophularia ningpoensis polysaccharide Download PDFInfo
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Abstract
The invention discloses scrophularia ningpoensis polysaccharide and a method for microwave extraction of scrophularia ningpoensis polysaccharide. The method comprises the following steps: (1) pulping or grinding crude medicinal materials, and then enzymatically hydrolyzing the medicinal materials with a complex enzyme preparation; (2) carrying out microwave-assisted extraction at the material-to-liquid ratio of 1 to (10-100) of scrophularia ningpoensis to water, centrifugally filtering so as to obtain an extracting solution, and carrying out intermittent vacuumizing and vacuum breaking on an extracting tank during extracting; (3) loading the scrophularia ningpoensis polysaccharide extracting solution on a macroporous adsorption resin chromatographic column, eluting with de-ionized water and collecting eluate; (4) concentrating the eluate in reduced pressure, adding ethanol, fully stirring and uniformly mixing, and standing by; and (5) freeze-drying. By effectively integrating enzymatic hydrolysis with the complex enzyme preparation, vacuum pulsating, microwave assistance, resin adsorption and other technologies, the method disclosed by the invention can significantly improve the extraction rate and the purity of the scrophularia ningpoensis polysaccharide and can shorten an extraction time, and the method is strong in market competitiveness and broad in application prospect; and the prepared scrophularia ningpoensis polysaccharide has a significant inhibitory effect on cancer cells.
Description
Technical field
The present invention relates to radix scrophulariae polysaccharide and adopt microwave-assisted from Chinese medicinal materials radix scrophulariae, extract the method for polysaccharide.
Background technology
Vegetable polysaccharides is an important topic in biochemical field, and become new study hotspot at home and abroad, the water-soluble polysaccharide wherein extracted from herbal medicine is particularly popular.Polysaccharide be linked by monose form, the polymer of the polymerization degree more than more than 100,000, vegetable polysaccharides is a kind of composition be prevalent in plant cell wall and cell, and at present hundreds of polysaccharide existing is from nature separation and Extraction out.Large quantity research proves, polysaccharide has the biological activity of its uniqueness, is the main active substances of many herbal medicine, has antibacterial, antiviral, antitumor, radioprotective, anti-ageing effect of waiting for a long time, and wherein especially becomes the focus of Recent study exploitation with immunoregulation effect.Add up according to the Chinese Medicines&Health Produts chamber of import and export trade, by pulling of cosmopolitian plant pharmaceutical market demand growth, China in 2011 plant milk extract outlet presents the two good momentum increased of output valency, and especially export average price and comparatively within 2010, have increased significantly, market outlook are boundless.
Radix scrophulariae has another name called radix scrophulariae, black ginseng, heavy platform, ghost Tibetan, deer intestines, Xuan Tai etc., and radix scrophulariae is the dry root of goatweed radix scrophulariae (Scrophularia ningpoensis Hems1.), is China's traditional Chinese medicine.Radix scrophulariae cold nature, taste is sweet, bitter, salty, returns lung, stomach, kidney channel, has effect of cool blood enriching yin, eliminating fire and detoxication.Modern pharmacological research shows, the Main Function of radix scrophulariae has anti-inflammatory, strengthens immunologic function, hypoglycemic, protect the liver, antitumor, antifatigue etc., in addition, radix scrophulariae also has certain effect to neural system, as depressed in neuroprotective, Hangzhoupro, improve memory etc.
Radix scrophulariae polysaccharide also has huge pharmacological action as a kind of important extract of radix scrophulariae, and radix scrophulariae polysaccharide has Scavenging activity in various degree to hydroxyl radical free radical, ultra-oxygen anion free radical and nitrite.Wang Hui etc. are to the research of radix scrophulariae water soluble component, result shows that radix scrophulariae polysaccharide has and extends mouse swimming time, reduce the rear mice serum urinary nitrogen content of motion, reduce the effect of the rear Mouse Blood lactic acid content of motion and increase Mouse Liver glycogen, radix scrophulariae polysaccharide is improved the vigor of SOD, GSH-Px in mouse aging body and reduces the effect of MDA content in mouse aging body.
Radix scrophulariae polysaccharide is as a kind of Medicines and Health Product, and its function is huge, but its extraction process need to improve.The extraction of current radix scrophulariae polysaccharide is mostly adopt canned single batch of extraction, and productive rate is low, and extraction time is longer, and Extraction solvent expends comparatively large, and operate more loaded down with trivial details, environmental pollution is more serious.Therefore, the extracting method of exploitation radix scrophulariae polysaccharide, simplifies the operation, reduces the loss to active polysaccharide material, shorten extracting cycle, and the yield and the purity that improve polysaccharide are significant.
Summary of the invention
For overcoming the deficiencies in the prior art, the present inventor, through lot of experiments, gropes and sums up a kind of microwave extract method of radix scrophulariae polysaccharide, utilizes present method can significantly improve the extraction yield of radix scrophulariae polysaccharide and purity, shortening extraction time, reduce extraction cost, increase economic efficiency.
For overcoming the deficiencies in the prior art, the present invention takes following technical scheme:
(1) raw-material process: no matter radix scrophulariae is dry wet, wet carrying out making beating process, be then crushed to 20-50 order if dry, the compound enzymic preparation adding water and quality of medicinal material 8/2000 to thousand/1000ths carries out enzymolysis, enzymolysis time 4-10 hour, hydrolysis temperature 40-50 DEG C; Prove, with the extraction yield that can significantly improve polysaccharide after ferment treatment through test.
The proportioning of described compound enzymic preparation is cellulase massfraction 50-70%, and polygalacturonase massfraction is 15-40%, and all the other are zytase, and the average enzyme of compound enzymic preparation is lived as 10000-30000 unit;
(2) microwave extraction: take water as solvent, the solid-liquid ratio of radix scrophulariae and water is 1:10-1:100, extracts radix scrophulariae polysaccharide with microwave assisting method, obtains extracting solution after centrifuging;
In said extracted process, step vacuumizing and vacuum breaker is carried out to extractor, utilize vacuum impulse reinforcing mass transfer.The mechanism of vacuum impulse reinforcing mass transfer is: moisture when vacuumizing in radix scrophulariae cell tissue can gasify because of decompression, cause radix scrophulariae cell tissue internal pressure to be greater than outside radix scrophulariae histocyte, the water having dissolved polysaccharide can be forced through radix scrophulariae cell tissue under differential pressure action; When vacuum breaker, radix scrophulariae histocyte external pressure is greater than pressure in radix scrophulariae histocyte, moisture outside radix scrophulariae histocyte enters rapidly radix scrophulariae histocyte inside and goes to dissolve polysaccharide because of differential pressure action, the alternately change of this radix scrophulariae histocyte external and internal pressure enhances moisture and is dissolved in the polysaccharide mass transfer in moisture, makes that the extraction efficiency of polysaccharide improves, residual rate reduction.
(3) chromatography: radix scrophulariae polysaccharide extraction liquid is loaded to macroporous adsorption resin chromatography post, adopts deionized water wash-out, collects elutriant;
(4) alcohol precipitation: after elutriant concentrating under reduced pressure, adds the abundant stirring and evenly mixing of ethanol, leaves standstill, collecting precipitation;
(5) lyophilize: redissolved by alcohol precipitation gained throw out use water, lyophilize obtains radix scrophulariae polysaccharide sterling.
In technique scheme, in step (2), the optimal technical scheme of microwave extraction is: microwave power is 800-1500W, and Extracting temperature is 60 DEG C-100 DEG C, and extraction time is 10-60 minute.
In technique scheme, in step (3), chromatographic step is preferably: by polysaccharide extraction liquid loading to ADS-17 macroporous adsorptive resins, with the speed of 1.5-2.5ml/min deionized water wash-out, collects elutriant.ADS-17 macroporous adsorptive resins is high to radix scrophulariae polysaccharide adsorption rate, and easy wash-out, regenerate also easier.
In technique scheme, in step (4), alcohol precipitation step is preferably: chromatography gained elutriant is evaporated to 4-6 mg/ml, adds 3-6 doubly 95 % ethanol, 3-5 DEG C of standing 12-72 hour, centrifugal collecting precipitation.
Radix scrophulariae polysaccharide the present invention obtained carries out infrared spectrum characterization, at 3415.31cm
-1appearance one broad peak in place's is the stretching vibration of O-H; At 3000-2800cm
-1absorption peak in region is the characteristic peak of carbohydrate, and radix scrophulariae polysaccharide is at 2924.52cm
-1the absorption peak at place is polysaccharide molecule C-H stretching vibration; 1617.02cm
-1the absorption peak at place is the C=0 stretching vibration of kharophen in radix scrophulariae polysaccharide, illustrates in radix scrophulariae polysaccharide containing uronic acid; 1265.07 and 1142.62cm
-1there is the absorption band of the asymmetric and symmetrical stretching vibration of the C-O of ester group-C at place.
Prove through function test, the radix scrophulariae polysaccharide that the present invention obtains has obvious restraining effect to cancer cells.
Compared with prior art, beneficial effect of the present invention is also:
Raw medicinal material utilizes compound enzymic preparation to carry out enzymolysis before extracting, and by the cell wall damage of radix scrophulariae, enables polysaccharide component Fast Stripping, improves the extraction yield of polysaccharide.
Compound enzymic preparation of the present invention is an optimum combination, and for the feature of radix scrophulariae cell walls main component, design cellulase massfraction 50-70%, polygalacturonase massfraction is 15-40%, and all the other are zytase, makes hydrolysis result best.
Take water as solvent, adopt the polysaccharide component of microwave radiation exaraction to radix scrophulariae to extract, effectively shorten extraction time, improve the extraction yield of effective constituent; ADS-17 macroporous adsorptive resins is high to radix scrophulariae polysaccharide adsorption rate, and selectivity is strong, and easy wash-out, regenerate easy.
In the leaching process of radix scrophulariae polysaccharide, carry out vacuumizing and vacuum breaker, utilize vacuum impulse reinforcing mass transfer, shorten the extraction yield of extraction time and polysaccharide further.
In sum, the present invention is by carrying out effective integration by above multinomial technology, and significantly improve the extraction yield of radix scrophulariae polysaccharide and purity, shortening extraction time, reduce extraction cost, improve economic benefit, the market competitiveness is strong, has a extensive future.
Accompanying drawing explanation
Fig. 1 is the radix scrophulariae polysaccharide infrared absorpting light spectra utilizing the present invention to obtain.
Embodiment
Now by reference to the accompanying drawings, the microwave extract method of radix scrophulariae polysaccharide is illustrated further.
Embodiment 1:
(1) raw-material process: no matter radix scrophulariae is dry wet, wet carrying out making beating process, be then crushed to 25 orders if dry, the compound enzymic preparation adding water and quality of medicinal material 3/1000ths carries out enzymolysis, enzymolysis time 9 hours, hydrolysis temperature 45 DEG C;
The proportioning of described compound enzymic preparation is cellulase massfraction 52%, and polygalacturonase massfraction is 30%, and all the other are zytase, and the average enzyme work of compound enzymic preparation is 18000 units;
(2) microwave extraction: take water as solvent, the solid-liquid ratio of radix scrophulariae and water is 1:12, extracts radix scrophulariae polysaccharide with microwave assisting method, obtains extracting solution after centrifuging;
In said extracted process, step vacuumizing and vacuum breaker is carried out to extractor, utilize vacuum impulse reinforcing mass transfer.
(3) chromatography: radix scrophulariae polysaccharide extraction liquid is loaded to macroporous adsorption resin chromatography post, adopts deionized water wash-out, collects elutriant;
(4) alcohol precipitation: after elutriant concentrating under reduced pressure, adds the abundant stirring and evenly mixing of ethanol, leaves standstill, collecting precipitation;
(5) lyophilize: redissolved by alcohol precipitation gained throw out use water, lyophilize obtains radix scrophulariae polysaccharide sterling.
In step (2), microwave power is 850W, and Extracting temperature is 65 DEG C, and extraction time is 30 minutes.
In step (3), by polysaccharide extraction liquid loading to ADS-17 macroporous adsorptive resins, with the speed of 1.6ml/min deionized water wash-out, collect elutriant.
In step (4), chromatography gained elutriant is evaporated to 4.2 mg/ml, adds 4 times of 95 % ethanol, 4 DEG C leave standstill 15 hours, centrifugal collecting precipitation.
Radix scrophulariae polysaccharide the present invention obtained carries out infrared spectrum characterization, at 3415.31cm
-1appearance one broad peak in place's is the stretching vibration of O-H; At 3000-2800cm
-1absorption peak in region is the characteristic peak of carbohydrate, and radix scrophulariae polysaccharide is at 2924.52cm
-1the absorption peak at place is polysaccharide molecule C-H stretching vibration; 1617.02cm
-1the absorption peak at place is the C=0 stretching vibration of kharophen in radix scrophulariae polysaccharide, illustrates in radix scrophulariae polysaccharide containing uronic acid; 1265.07 and 1142.62cm
-1there is the absorption band of the asymmetric and symmetrical stretching vibration of the C-O of ester group-C at place.
Prove through function test, the radix scrophulariae polysaccharide that the present invention obtains has obvious restraining effect to cancer cells.
embodiment 2:
(1) raw-material process: no matter radix scrophulariae is dry wet, wet carrying out making beating process, be then crushed to 45 orders if dry, the compound enzymic preparation adding water and quality of medicinal material 7/1000ths carries out enzymolysis, enzymolysis time 4.5 hours, hydrolysis temperature 48 DEG C;
The proportioning of described compound enzymic preparation is cellulase massfraction 67%, and polygalacturonase massfraction is 18%, and all the other are zytase, and the average enzyme work of compound enzymic preparation is 27000 units;
(2) microwave extraction: take water as solvent, the solid-liquid ratio of radix scrophulariae and water is 1:91, extracts radix scrophulariae polysaccharide with microwave assisting method, obtains extracting solution after centrifuging;
In said extracted process, step vacuumizing and vacuum breaker is carried out to extractor, utilize vacuum impulse reinforcing mass transfer.
(3) chromatography: radix scrophulariae polysaccharide extraction liquid is loaded to macroporous adsorption resin chromatography post, adopts deionized water wash-out, collects elutriant;
(4) alcohol precipitation: after elutriant concentrating under reduced pressure, adds the abundant stirring and evenly mixing of ethanol, leaves standstill, collecting precipitation;
(5) lyophilize: redissolved by alcohol precipitation gained throw out use water, lyophilize obtains radix scrophulariae polysaccharide sterling.
In step (2), microwave power is 1400W, and Extracting temperature is 90 DEG C, and extraction time is 50 minutes.
In step (3), by polysaccharide extraction liquid loading to ADS-17 macroporous adsorptive resins, with the speed of 2.3ml/min deionized water wash-out, collect elutriant.
In step (4), chromatography gained elutriant is evaporated to 5.8 mg/ml, adds 4.8 times of 95 % ethanol, 4.7 DEG C leave standstill 15 hours, centrifugal collecting precipitation.
Radix scrophulariae polysaccharide the present invention obtained carries out infrared spectrum characterization, at 3415.31cm
-1appearance one broad peak in place's is the stretching vibration of O-H; At 3000-2800cm
-1absorption peak in region is the characteristic peak of carbohydrate, and radix scrophulariae polysaccharide is at 2924.52cm
-1the absorption peak at place is polysaccharide molecule C-H stretching vibration; 1617.02cm
-1the absorption peak at place is the C=0 stretching vibration of kharophen in radix scrophulariae polysaccharide, illustrates in radix scrophulariae polysaccharide containing uronic acid; 1265.07 and 1142.62cm
-1there is the absorption band of the asymmetric and symmetrical stretching vibration of the C-O of ester group-C at place.
Prove through function test, the radix scrophulariae polysaccharide that the present invention obtains has obvious restraining effect to cancer cells.
Claims (5)
1. a microwave extract method for radix scrophulariae polysaccharide, is characterized in that comprising following step:
(1) raw-material process: no matter radix scrophulariae is dry wet, wet carrying out making beating process, be then crushed to 20-50 order if dry, the compound enzymic preparation adding water and quality of medicinal material 8/2000 to thousand/1000ths carries out enzymolysis, enzymolysis time 4-10 hour, hydrolysis temperature 40-50 DEG C;
The proportioning of described compound enzymic preparation is cellulase massfraction 50-70%, and polygalacturonase massfraction is 15-40%, and all the other are zytase, and the average enzyme of compound enzymic preparation is lived as 10000-30000 unit;
(2) microwave extraction: take water as solvent, the solid-liquid ratio of radix scrophulariae and water is 1:10-1:100, extracts radix scrophulariae polysaccharide with microwave assisting method, obtains extracting solution after centrifuging, in leaching process, step vacuumizing and vacuum breaker is carried out to extractor, utilize vacuum impulse reinforcing mass transfer;
(3) chromatography: radix scrophulariae polysaccharide extraction liquid is loaded to macroporous adsorption resin chromatography post, adopts deionized water wash-out, collects elutriant;
(4) alcohol precipitation: after elutriant concentrating under reduced pressure, adds the abundant stirring and evenly mixing of ethanol, leaves standstill, collecting precipitation;
(5) lyophilize: redissolved by alcohol precipitation gained throw out use water, lyophilize obtains radix scrophulariae polysaccharide sterling.
2. the microwave extract method of radix scrophulariae polysaccharide according to claim 1, it is characterized in that: in microwave extraction step, microwave power is 800-1500W, and Extracting temperature is 60 DEG C-100 DEG C, and extraction time is 10-60 minute.
3. the microwave extract method of radix scrophulariae polysaccharide according to claim 1, is characterized in that: in chromatographic step, by polysaccharide extraction liquid loading to ADS-17 macroporous adsorptive resins, with the speed of 1.5-2.5ml/min deionized water wash-out, collects elutriant.
4. the microwave extract method of radix scrophulariae polysaccharide according to claim 1, is characterized in that: in alcohol precipitation step, chromatography gained elutriant is evaporated to 4-6 mg/ml, adds 3-6 doubly 95 % ethanol, 3-5 DEG C of standing 12-72 hour, centrifugal collecting precipitation.
5. the radix scrophulariae polysaccharide sterling that method obtains according to claim 1 or 2 or 3 or 4, is characterized in that: radix scrophulariae polysaccharide has obvious restraining effect to cancer cells.
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| CN108837061A (en) * | 2018-07-21 | 2018-11-20 | 安溪县凤城建金产品外观设计服务中心 | The extraction process of effective component in a kind of watt of reed |
| CN112794929A (en) * | 2021-01-25 | 2021-05-14 | 西藏天虹科技股份有限责任公司 | Highland barley beta-glucan extraction process based on ultrasonic assistance |
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