CN102241786B - Preparation method and application of selenium enriched puerarin polysaccharide - Google Patents
Preparation method and application of selenium enriched puerarin polysaccharide Download PDFInfo
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Abstract
The invention discloses a preparation method and application of selenium enriched puerarin polysaccharide, and belongs to the technical field of health products. The preparation method comprises the following steps of: enriching selenium in kudzu root culture, namely preparing sodium selenite into an aqueous solution and performing foliage spray on the kudzu root for enriching the selenium; crushing the kudzu root into powder with the granularity of 80 to 120 meshes; extracting by using a 3 mass percent complex enzyme solution in an amount which is 10 to 30 times weight of the kudzu root, refluxing at the temperature of between 40 and 50DEG C for 2 to 4 hours, and extracting for 2 to 4 times; centrifuging extracting solution, and concentrating supernate to reach certain concentration, wherein the concentration multiple is 8 to 20 times; removing protein by using trichloroacetic acid, adding ethanol for precipitation, and making the ethanol concentration in the concentrated liquor of the polysaccharide 60 to 85 percent; performing secondary ethanol precipitation on the supernate; and performing freeze drying on a precipitate to obtain the selenium enriched puerarin polysaccharide. By the method, selenium enriched kudzu root can be produced, and the method is simple and convenient, low in cost and good in effect. The selenium enriched puerarin polysaccharide can serve as an antioxidant functional food, and also can serve as a human selenium supplement food.
Description
Technical field
The present invention relates to a kind of preparation method of rich selenium pueraria polysaccharide and as the application of antioxidant and replenishing selenium food, belong to the health food technology field.
Background technology
Selenium be the human and animal essential trace element, have remove lipid peroxide, improve immunizing power, anticancer, remove the critical functions such as heavy metallic poison and continuity be old and feeble, be called as " kindling material of life ".The selenium daily intaking amount standard that Chinese Soclety of Nutrition is recommended is 20 ~ 50 μ g.Selenium deficiency causes the body metabolism obstacle often, and is easily diseases induced.Have in the world many areas to lack selenium, there is 72% the scarce selenium in county in China, has approximately 300,000,000 people to be subject to lacking selenium and threatens, and therefore, replenishing selenium becomes the major issue that lacks selenium area urban and rural residents health.Studies show that the easiest selenium that is absorbed by the body is the organoselenium in the plant, therefore, the vegetable food product of selenium is rich in exploitation, makes the selenium element by plant absorbing, is converted into organoselenium through biological action, and eating for people is the best method of mending selenium again.About the existing patent report of the research of the rich selenium of plant.Chinese patent CN101292615 discloses " method of growing nuisance free selenium-rich rice by using biological technique " on October 29th, 2008, has obtained non-harmful Selenium rich rice.Chinese patent CN101496476 discloses " selenium-rich chrysanthemum " on July 21st, 2009, cultivate to obtain a kind of selenium-rich chrysanthemum.Chinese patent CN101779542A discloses " production method of organic selenium-rich Huangshan Maofeng tea " on July 21st, 2010, has produced a kind of Huangshan Maofeng tea that is rich in organoselenium.Elegant jessamine is the perennial defoliation liana of pulse family, has medicine-food two-purpose concurrently and is worth.The root of kudzu vine that Chinese Pharmacopoeia records is the dry root of elegant jessamine, and it is very wide to distribute in China, and except Tibet and Xinjiang, all there is distribution all the other each provinces and regions, its medicinal Shennong's Herbal that is recorded in the earliest.The root of kudzu vine contains the various active composition, and wherein starch content is high, and the fresh root of kudzu vine contains 20%～30% starch, and dried pueraria starch content reaches 35%～40%; Soluble dietary fibre content is 39.53% in the elegant jessamine, and insoluble dietary fibre content is 10.54%, and robust fibre is 14.19%, and xylogen is 16.3l%.Radix Puerariae isoflavone is the general name of the flavonoid compound in the root of kudzu vine, it is the main active chemical of the root of kudzu vine, wherein take osajin compositions such as puerarin, daidzin and daidzeins as main, can be used for Cardiovarscular, stenocardia, irregular pulse and gynaecopathia etc.At present to the research of the root of kudzu vine mainly for the osajin composition, few to other activeconstituentss such as polysaccharide and bioactive research thereof.Chinese patent CN101736052A discloses " technique of high efficiency extraction pueraria starch and root of kudzu vine total isoflavone " on June 16th, 2010, and high efficiency extraction obtains powder of Radix Puerariae and root of kudzu vine total isoflavone from the root of kudzu vine.Chinese patent CN101805410A discloses " processing method of a kind of comprehensive production Radix Puerariae total flavones, puerarin and pueraria starch ", simultaneous extraction root of kudzu vine total isoflavone, puerarin and pueraria starch from the root of kudzu vine on August 18th, 2010.The inventor adopts the blade face to execute the selenium mode root of kudzu vine to be carried out rich selenium, obtain the rich selenium root of kudzu vine, then extracts and obtains rich selenium pueraria polysaccharide, has studied the anti-oxidant activity of rich selenium pueraria polysaccharide.The rich selenium of the relevant root of kudzu vine and the anti-oxidant activity of rich selenium pueraria polysaccharide research so far there are no domestic and foreign literature report also has no corresponding patent disclosure.
Detailed content of the present invention:
The objective of the invention is the deficiency that exists in order to solve the above-mentioned background technology, provide a kind of preparation method of rich selenium pueraria polysaccharide and as the application of antioxidant and replenishing selenium food etc.The preparation method of rich selenium pueraria polysaccharide comprises selenium-rich method in the root of kudzu vine cultivation and the extracting method of rich selenium pueraria polysaccharide.
The preparation method of a kind of rich selenium pueraria polysaccharide of the present invention, the method is carried out according to following step:
One, the selenium-rich method in the root of kudzu vine cultivation
(1) is mixed with the aqueous solution of 10 ~ 40 mg/L with Sodium Selenite;
(2) adopt foliage-spray that the root of kudzu vine is carried out rich selenium;
(3) executing the selenium time is July ~ September, and the cycle is 20 ~ 25 days.
Two, the extracting method of rich selenium pueraria polysaccharide
(1) root of kudzu vine is pulverized: powder of Radix Puerariae is broken to 80 ~ 120 orders;
(2) polysaccharide extracts: 10 ~ 30 times of amounts composite enzyme solution of containing quality amount concentration 3% is extracted by weight, and 2 ~ 4 h that reflux under 40 ~ 50 ℃ temperature extract 2 ~ 4 times;
(3) extracting solution is centrifugal, and centrifugal speed is 3000 ~ 4000 r/min; Get supernatant concentration after centrifugal to finite concentration, cycles of concentration is 8 ~ 20 times; Remove albumen with trichoroacetic acid(TCA);
(4) add the ethanol precipitation, so that alcohol concn is 60 ~ 85% in the concentrated solution of polysaccharide; Supernatant liquor carries out the secondary alcohol precipitation;
(5) the gained throw out adopts freeze-drying dry, and dry 24 ~ 36 h get rich selenium pueraria polysaccharide.
Prozyme described in the step 2 of the present invention is cellulase in mass: the mixture of polygalacturonase: pancreatin=2:2:1.
The application of above-mentioned rich selenium pueraria polysaccharide can be used as antioxidant or is prepared into the food of replenishing selenium.
The invention has the advantages that: adopt method of the present invention to produce and obtain the rich selenium root of kudzu vine, method is easy, and cost is low, and is effective; Can produce the rich selenium pueraria polysaccharide of acquisition take the rich selenium root of kudzu vine as raw material, selenium content wherein is more than 5 times of selenium content in the common pueraria polysaccharide; The anti-oxidant activity of rich selenium pueraria polysaccharide is good, with Vc quite.Therefore rich selenium pueraria polysaccharide of the present invention both can be used as the antioxidant functional food except the medicinal and health-care effect that keeps common pueraria polysaccharide, can be used as again replenishing selenium food.
Description of drawings
Fig. 1 is that rich selenium pueraria polysaccharide is to the DPPH free radical scavenging activity;
Fig. 2 is that rich selenium pueraria polysaccharide is to Scavenging action to hydroxyl free radical;
Fig. 3 is that rich selenium pueraria polysaccharide is to ABTS
+Free radical scavenging activity.
Embodiment
Rich selenium sample is measured selenium content with By Hg-afs among the present invention.
The mensuration of the anti-oxidant activity of rich selenium pueraria polysaccharide comprises among the present invention: rich selenium pueraria polysaccharide is to DPPH free radical, hydroxyl radical free radical, ABTS
+The removing effect of free radical.
Adopt among the present invention and remove DPPH free radical, hydroxy radical qiao, ABTS
+The anti-oxidant activity of rich selenium pueraria polysaccharide is investigated in test.Result of study shows that the anti-oxidant activity of rich selenium pueraria polysaccharide is good, and is suitable with Vc.Therefore rich selenium pueraria polysaccharide of the present invention can be used as oxidation resistant functional foodstuff.
Employed cellulase among the present invention (traditional Chinese medicines company product), enzyme 10000 LEU/g alive; Polygalacturonase (traditional Chinese medicines company product), enzyme 10000 LEU/g alive; Pancreatin (Jining of Shandong Province and U.S. biotechnology company limited product), enzyme 10000 LEU/g that live.
In order to be illustrated more clearly in content of the present invention, be described as follows with specific embodiment, specific embodiment does not limit context of the present invention.
Embodiment 1
This test is carried out foliage-spray (by 100 mL/m with 10 mg/L sodium selenite aqueous solutions to the root of kudzu vine
2), to carry out spraying the first time July in busy season in root of kudzu vine growth, every interval sprayed once in 20 days, altogether sprayed 4 times.Gather in the crops the rich selenium root of kudzu vine March in Second Year, clean, chopping in 60 ℃ of oven dry, is pulverized, and after 120 mesh sieves divide, dries to constant weight in 60 ℃ of baking ovens again, gets the rich selenium root of kudzu vine.Take by weighing the rich selenium root of kudzu vine 20 g, the prozyme of adding mass concentration 3% [the W(cellulase): the W(polygalacturonase): the W(pancreatin)=2:2:1], vat liquor pH is 7.0, volume is 500 mL, temperature is 40 ℃, lixiviate under the extraction times 2.5 h conditions, vat liquor behind the concentrating under reduced pressure, removes albumen with trichoroacetic acid(TCA) after filtration, add 95% ethanol alcohol precipitation, 12 h according to volume ratio 1:4, centrifugal, supernatant liquor carries out the secondary alcohol precipitation, and is centrifugal, lyophilize gets rich selenium pueraria polysaccharide sample powder 1.904 g, and wherein polysaccharide content is 44.17%.Extracting the common root of kudzu vine with method obtains prozyme and carries pueraria polysaccharide.By Hg-afs records the common root of kudzu vine and the rich selenium root of kudzu vine is respectively 0.0527 ± 0.0052 μ g/g and 0.1218 ± 0.0112 μ g/g through the Wet selenium content, and selenium content is more than 2 times of selenium content in the common root of kudzu vine in the visible rich selenium root of kudzu vine; By Hg-afs records common pueraria polysaccharide and rich selenium pueraria polysaccharide is respectively 0.1724 ± 0.0163 μ g/g and 0.4653 ± 0.0338 μ g/g through the Wet selenium content, and selenium content is nearly 3 times of selenium content in the common pueraria polysaccharide in the visible rich selenium pueraria polysaccharide.
Embodiment 2
This test is carried out foliage-spray (by 100 mL/m with 20 mg/L sodium selenite aqueous solutions to the root of kudzu vine
2), all the other steps get the rich selenium root of kudzu vine with the selenium-rich method in the root of kudzu vine cultivation of example 1.Take by weighing the rich selenium root of kudzu vine 20 g, the prozyme of adding mass concentration 3% [the W(cellulase): the W(polygalacturonase): the W(pancreatin)=2:2:1], vat liquor pH is 7.0, volume is 600 mL, temperature is 50 ℃, lixiviate under the extraction times 3 h conditions, and all the other steps are with the extracting method of the rich selenium pueraria polysaccharide of example 1, get rich selenium pueraria polysaccharide sample powder 1.912 g, wherein polysaccharide content is 42.89%.It is 0.2086 ± 0.0205 μ g/g through the Wet selenium content that By Hg-afs records the rich selenium root of kudzu vine, and selenium content is nearly 4 times of selenium content in the common root of kudzu vine in the visible rich selenium root of kudzu vine; By Hg-afs records rich selenium pueraria polysaccharide and is respectively 0.8885 ± 0.0238 μ g/g through the Wet selenium content, and selenium content is more than 5 times of selenium content in the common pueraria polysaccharide in the visible rich selenium pueraria polysaccharide.
Embodiment 3
This test is carried out foliage-spray (by 100 mL/m with 40 mg/L sodium selenite aqueous solutions to the root of kudzu vine
2), all the other steps get the rich selenium root of kudzu vine with the selenium-rich method in the root of kudzu vine cultivation of example 1.Take by weighing the rich selenium root of kudzu vine 20 g, the prozyme of adding mass concentration 3% [the W(cellulase): the W(polygalacturonase): the W(pancreatin)=2:2:1], vat liquor pH is 7.0, volume is 600 mL, temperature is 50 ℃, lixiviate under the extraction times 4 h conditions, and all the other steps are with the extracting method of the rich selenium pueraria polysaccharide of example 1, get rich selenium pueraria polysaccharide sample powder 1.825 g, wherein polysaccharide content is 42.58%.By Hg-afs records the rich selenium root of kudzu vine, and to record selenium content through Wet be 0.2408 ± 0.0238 μ g/g, and selenium content is more than 4 times of selenium content in the common root of kudzu vine in the visible rich selenium root of kudzu vine; It is 1.1213 ± 0.1138 μ g/g through the Wet selenium content that By Hg-afs records rich selenium pueraria polysaccharide, and selenium content is more than 6 times of selenium content in the common pueraria polysaccharide in the visible rich selenium pueraria polysaccharide.
Test the antioxidant activity tests of a rich selenium pueraria polysaccharide
By removing DPPH free radical, hydroxy radical qiao and ABTS
+ The anti-oxidant activity of given the test agent is investigated in the free radical test
Rich selenium pueraria polysaccharide is removed DPPH free radical, hydroxy radical qiao and ABTS
+ Free Radical
1, experiment material
1.1 medicine and reagent
Rich selenium pueraria polysaccharide: the aqueous solution that is mixed with crude drug concentration 0.5 mg/mL is for subsequent use;
DPPH solution: the ethanol solution that is mixed with 0.2 mmol/L;
Phenanthroline solution: the ethanol solution that is mixed with 0.75 mmol/L;
Phosphate buffer soln: the buffered soln that is mixed with 0.2 mmol/L pH7.40;
Copperas solution: the ethanol solution that is mixed with 0.75 mmol/L;
Hydrogen peroxide: be configured to 0.01% hydrogen peroxide;
ABTS
+ Solution: the solution that is mixed with 0.2 mmol/L;
1.2 instrument
TU-1800 ultraviolet-visible pectrophotometer: Beijing Puxi General Instrument Co., Ltd;
BS124S electronic balance: Beijing Sai Duolisi instrument system company limited;
Freeze drier FD-1C-50: Beijing rich doctor health laboratory apparatus company limited;
Rotatory evaporator RE-52C: Yuhua Instrument Co., Ltd., Gongyi City;
2, experimental technique
2.1 rich selenium pueraria polysaccharide is removed the DPPH Free Radical
Get respectively the sample of 2 mL different concns in test tube, DPPH solution 2 mL that add 0.2 mmol/L, mix, after 30 min are reacted in the dark place, measure its absorbancy (Ai) at 517 nm places, replace sample determination blank absorbency (Ao) with solvent, replace DPPH to measure absorbancy (Aj) with dehydrated alcohol.
Clearance rate S(%)=[1-(Ai-Aj)/Ao] * 100%
2.2 rich selenium pueraria polysaccharide scavenging hydroxyl free radical
Get 1 mL phenanthroline ethanol solution, add phosphate buffered saline buffer and the 1 mL distilled water of 2 ml, 0.2 mL/L pH 7.4, after fully mixing, add 1 mL0.75 mmoL/L FeSO
4Mix, add again 1 mL 0.01%H
2O
2, 37 ℃ of heating in water bath 60 min, 536 nm places survey light absorption value (A damage), and 1 mL sample liquid replaces 1 mL distilled water to record the light absorption value (A sample) of sample, and 1 mL distilled water replaces 1 mL hydrogen peroxide to record light absorption value (A is not).
Clearance rate S(%)=(A sample-A decrease)/(A not-A decrease) * 100%
2.3 rich selenium pueraria polysaccharide is removed ABTS
+ Free Radical
Get respectively sample and the 3.8 mL ABTS of 2 mL different concns
+ Solution mixes, behind 6 min in 732 nm places mensuration absorbancy.Replace sample to get Ao with sample solvent, replace ABTS solution to get Aj with damping fluid.
Clearance rate S(%)=[1-(Ai-Aj)/Ao] * 100%
3, experimental result
Fig. 1 be rich selenium pueraria polysaccharide and Vc to the removing effect of DPPH free radical, rich pueraria polysaccharide is when 2 mg/mL, it reaches 80% to the DPPH free radical scavenging activity, its IC
50Be the IC of 0.9699 mg/mL(Vc
50Be 0.4563 mg/mL), visible rich pueraria polysaccharide is effective to the removing of DPPH free radical.Fig. 2 be rich selenium pueraria polysaccharide and Vc to the removing effect of hydroxyl radical free radical, rich pueraria polysaccharide is when 1.2 mg/mL, it reaches 73% to Scavenging action to hydroxyl free radical, its IC
50Be the IC of 0.6158 mg/mL(Vc
50Be 0.3128 mg/mL), visible rich pueraria polysaccharide is effective to the removing of hydroxyl radical free radical.Fig. 3 is that rich selenium pueraria polysaccharide and Vc are to ABTS
+The removing effect of free radical, rich selenium pueraria polysaccharide is when 1.2 mg/mL, and it is to ABTS
+Free radical scavenging activity reaches 62%, its IC
50Be the IC of 0.6732 mg/mL(Vc
50Be 0.3835 mg/mL), visible rich pueraria polysaccharide is to ABTS
+The removing of free radical is effective.
4, conclusion
Rich selenium pueraria polysaccharide is to DPPH free radical, hydroxy radical qiao and ABTS
+The removing of free radical is effective, with Vc quite.
Claims (3)
1. the preparation method of a rich selenium pueraria polysaccharide is characterized in that carrying out according to following step:
One, the selenium-rich method in the root of kudzu vine cultivation:
(1) is mixed with the aqueous solution of 10 ~ 40 mg/L with Sodium Selenite;
(2) adopt foliage-spray that the root of kudzu vine is carried out rich selenium;
(3) executing the selenium time is July ~ September, and the cycle is 20 ~ 25 days;
Two, the extracting method of rich selenium pueraria polysaccharide:
(1) root of kudzu vine is pulverized: powder of Radix Puerariae is broken to 80 ~ 120 orders;
(2) polysaccharide extracts: 10 ~ 30 times of amounts composite enzyme solution of containing mass concentration 3% is extracted by weight, and 2 ~ 4 h that reflux under 40 ~ 50 ℃ temperature extract 2 ~ 4 times;
(3) extracting solution is centrifugal, and centrifugal speed is 3000 ~ 4000 r/min; Get supernatant concentration after centrifugal to finite concentration, cycles of concentration is 8 ~ 20 times; Remove albumen with trichoroacetic acid(TCA);
(4) add the ethanol precipitation, so that alcohol concn is 60 ~ 85% in the concentrated solution of polysaccharide; Supernatant liquor carries out the secondary alcohol precipitation;
(5) the gained throw out adopts freeze-drying dry, and dry 24 ~ 36 h get rich selenium pueraria polysaccharide;
Prozyme described in the step 2 is cellulase in mass: the mixture of polygalacturonase: pancreatin=2:2:1.
2. the application of rich selenium pueraria polysaccharide claimed in claim 1 is as the application of antioxidant.
3. the application of rich selenium pueraria polysaccharide claimed in claim 1 is prepared into the application of the food of replenishing selenium.
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Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102805131A (en) * | 2012-07-27 | 2012-12-05 | 安徽四海食品有限公司 | Health-care flour containing kudzuvine root polysaccharide and preparation method of health-care flour |
| CN102835240A (en) * | 2012-09-03 | 2012-12-26 | 苏延友 | Cultivation method of selenium-enriched Codonopsis lanceolata, as well as extraction method and application of selenium-containing polysaccharide of selenium-enriched Codonopsis lanceolata |
| CN103609974B (en) * | 2013-11-09 | 2015-06-17 | 安徽金钗石斛有限公司 | Kudzuvine root health-care rice crust |
| CN103652886B (en) * | 2013-11-30 | 2015-07-01 | 广西桂林天然食品有限公司 | High-protein nutritious and health-care compound rice containing konjac cereals rich in selenium, zinc and calcium |
| CN103689344B (en) * | 2013-11-30 | 2015-08-26 | 广西桂林天然食品有限公司 | A kind of high protein selenium-rich, Fu Xin, rich calcium root of kudzu vine coarse cereals composite nutritional healthcare rice |
| CN105063129A (en) * | 2015-08-07 | 2015-11-18 | 广西南宁派腾科技有限公司 | Ultrasonic extraction process of kudzuvine root fructose |
| CN106279446B (en) * | 2016-08-29 | 2017-09-01 | 唐汉军 | The method that starch, fibrous polysaccharaide and water-soluble isoflavones are extracted from the root of kudzu vine |
| CN106749719A (en) * | 2016-12-05 | 2017-05-31 | 江苏大学 | A kind of selenium-rich grifola frondosus selenium polysaccharide and its preparation method and application |
| CN106916233A (en) * | 2017-04-26 | 2017-07-04 | 湖北省农业科学院中药材研究所 | A kind of preparation method of RADIX PUERARIAE polysaccharide |
| CN108484787B (en) * | 2018-04-11 | 2020-10-23 | 钦州学院 | Preparation method of moringa oleifera selenium-rich polysaccharide |
| CN109824795A (en) * | 2019-03-18 | 2019-05-31 | 新昌县大船畈生物科技有限公司 | The extracting method of selenium-rich Moringa |
| CN110028595A (en) * | 2019-04-19 | 2019-07-19 | 安徽工程大学 | A kind of Deproteinated method of pueraria polysaccharide |
| CN110250522A (en) * | 2019-07-14 | 2019-09-20 | 乐山合亩农业生物科技有限公司 | A kind of production method of organism organic selenium |
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| CN101020720B (en) * | 2006-12-28 | 2010-12-15 | 宋桃森 | Process of extracting pueraria polysaccharide |
| CN101570581A (en) * | 2009-06-10 | 2009-11-04 | 华中农业大学 | Method for comprehensively extracting pueraria polysaccharide and flavone |
| CN101704904B (en) * | 2009-11-13 | 2012-05-23 | 江苏大学 | Preparation method and application of kudzu polysaccharide extract |
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