CN102030834B - Method for extracting and preparing camellia polysaccharide from camellia and application of camellia polysaccharide - Google Patents
Method for extracting and preparing camellia polysaccharide from camellia and application of camellia polysaccharide Download PDFInfo
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- 206010012601 diabetes mellitus Diseases 0.000 claims abstract description 11
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a method for extracting and preparing camellia polysaccharide from a camellia and an application of the prepared camellia polysaccharide. The method comprises the following steps: extracting with preferable water or enzyme-containing water, carrying out alcohol precipitation on the extracting solution, separating, drying the separated precipitate to obtain camellia coarse polysaccharide, decoloring and deproteinizing the camellia coarse polysaccharide by resin or a chemical method, and purifying with gel or an ultrafiltration membrane to obtain the camellia polysaccharide. Simultaneously, the animal experiment proves that the camellia polysaccharide has the effect of preventing and treating diabetes.
Description
Technical field
The invention belongs to food, protective foods or medicine preparing technical field, particularly from camellia, extract and prepare the method for camellia polysaccharide, and relate to the medicinal use of this camellia polysaccharide.
Background technology
Camellia is one of reproductive organ of tea tree.Tea tree starts bud differentiation annual May, and bloom 9～October, many 100～110 days of florescence.The south China tea district tea tree florescence is longer, can extend to 2～March of next year, and the even annual visible camellia in indivedual areas is open.Tea district, the whole nation can produce more than 300 ten thousand tons of camellias every year, and resource is quite abundant.
Research shows, contains multiple beneficial composition and the active substances such as abundant polysaccharide, protein, amino acid, VITAMIN in camellia, and human body is had to the effects such as hypoglycemic, lipopenicillinase, removing toxic substances, anticancer, nourishing, beauty treatment.Practical proof, the anti-oxidant function of camellia can match in excellence or beauty with internationally recognized oxidation resistant plant Rosmarinus officinalis.
Yet the at present utilization research of people to tea, relate generally to be tealeaves in the effect of the aspects such as food, medical science and body-care, and the exploitation research of Tea Flower is not yet in full swing, a large amount of camellias can only be mixed with earth by self falling back to nature mutually.Until 2002, the famous Tea Science expert Xu Ji outstanding person of China proposes to develop Tea Flower resource, makes full use of existing camellia resource, carries out deep processing, develops the product useful to the mankind.If make full use of camellia resource, extract its effective constituent, not only there is larger social benefit, and can greatly increase economic efficiency.
Summary of the invention
One of technical problem to be solved by this invention is to provide a kind of and take camellia and as raw material extracts, prepare the method for camellia Crude polysaccharides and obtain camellia polysaccharide by the method.
Two of technical problem to be solved by this invention is to provide a kind of purification process of camellia Crude polysaccharides.
Three of technical problem to be solved by this invention is to provide the purposes of camellia polysaccharide.
The camellia of take as first aspect present invention is prepared the method for camellia Crude polysaccharides as raw material extracts, specifically comprise the steps:
A, by dry camellia after crushed, adds water extraction after 30 minutes, filters or centrifugation obtains solid precipitation and extracting solution;
B, concentrated extracting solution to density are greater than after 1.1Kg/L, add 2 times with upper volume lower alcohol, to staticly settle more than 12 hours, and supernatant liquor reclaims rudimentary pure recycling, obtain camellia Crude polysaccharides after drying precipitate.
Above-mentioned, take camellia and prepare in the method for camellia Crude polysaccharides as raw material extracts, in described water, can add zymin, wherein the mass ratio of enzyme and water is 0.1～1.2: 100.Described enzyme comprises any destruction camellia fibrous textures such as polygalacturonase, cellulase, is conducive to polysaccharide and extracts efficient, to improve yield single enzyme preparation or its two or more zymin mixture.
Above-mentioned, take camellia and prepare in the method for camellia Crude polysaccharides as raw material extracts, described extraction can be carried out at normal temperature to 100 ℃.
Above-mentioned, take camellia and prepare in the method for camellia Crude polysaccharides as raw material extracts, described extraction includes but not limited to: lixiviate, boiling water extraction, supersound extraction, microwave extraction or and anyly take the extraction process that water is medium.
Above-mentioned, take camellia and prepare in the method for camellia Crude polysaccharides as raw material extracts, described lower alcohol is preferably ethanol.
Above-mentioned, take camellia and prepare in the method for camellia Crude polysaccharides as raw material extracts, concentrating under reduced pressure at 55 ℃～100 ℃ of described simmer down tos, preferably 75 ℃.
Above-mentioned, take camellia and prepare in the method for camellia Crude polysaccharides as raw material extracts, describedly dryly comprise at 55 ℃～100 ℃ that reduced vacuum is dry, frozen drying or spraying dry.
Purification process as the camellia Crude polysaccharides of second aspect present invention, specifically comprises following method:
A, use macroporous adsorbent resin or ion exchange resin decolour and remove portion protein to camellia Crude polysaccharides; Or use chemical deproteinated and decolouring to process; Or use filter membrane that molecular weight is less than to 10,000 camellia Crude polysaccharides, monose, inorganic salt and other Impurity removals and obtain tea polysaccharide;
The resulting tea polysaccharide of b, step a, is used the filter membrane of different molecular weight to obtain the polysaccharide molecular weight different tea polysaccharide that distributes;
C, the resulting tea polysaccharide of use gel-purified step a, the separated different molecular weight that obtains after purifying
The camellia polysaccharide of the camellia polysaccharide of distribution range and different electric charge, ionic strength.
As a third aspect of the present invention, be the purposes of camellia polysaccharide, the camellia polysaccharide described in it can be used for preparing prevention and treatment diabetes.
Camellia polysaccharide of the present invention is through the modeling of tetraoxypyrimidine SD rat, and gavage, after 30 days, is compared with model group rat, and blood sugar concentration has significance and reduces.Results of animal proves, the effect that camellia polysaccharide has the diabetes of preventing and treating that obtains.
Extracting and preparing technique of the present invention is simple, pollution-free.Be applicable to large-scale production.
Embodiment
Embodiment 1
Dry camellia 10Kg feeds intake, and pulverizes 16 mesh sieves, adds 80L volume water, stir, and extract at room temperature 30 minutes, 4500 revs/min are centrifugal, make solid-liquid separation, collect extracting solution.Solid precipitation camellia adds 60L water again, stirs extract at room temperature 30 minutes, and 4500 revs/min are centrifugal, make solid-liquid separation, extracting solution and extracting solution merging for the first time, and solid precipitation camellia discards.
Extracting solution after merging is evaporated to density and is greater than 1.2Kg/L under 75 degrees Celsius, adds 4 times of volume ethanol precipitations after cooling 12 hours.Supernatant liquor ethanol reclaims, and throw out is dry 75 degrees Celsius of lower reduced vacuum.Obtain camellia Crude polysaccharides 1.68Kg.
Embodiment 2
Dry camellia 10Kg feeds intake, and pulverizes 16 mesh sieves, adds 80L volume water, stirs, and microwave 600w extracts 10 minutes, and Plate Filtration is collected extracting solution.Solid camellia adds 60L water again, and microwave 600w extracts 10 minutes, and Plate Filtration, makes solid-liquid separation, extracting solution and extracting solution merging for the first time, and solid precipitation camellia discards.
After merging, extracting solution is evaporated to density and is greater than 1.1Kg/L under 75 degrees Celsius, adds 4 times of volume ethanol precipitations 12 hours after cooling.Supernatant ethanol reclaims, vacuum freezedrying.Obtain camellia Crude polysaccharides 211g.
Embodiment 3
Gained camellia Crude polysaccharides in embodiment 1, with after water dissolution, ion exchange resin RJA on 3 times of column volume flow velocitys per hour (purchased from the Shanghai fast science equipment company limited that rubs) post, after loading, wash-out obtains tea polysaccharide, yield 82%, decolorizing effect is good.
Embodiment 4
In embodiment 3, gained camellia polysaccharide, through DEAE Sepharose Fast Flow gel-purified, obtains two simple spikes after separation, and the former is neutral sugar peak, and the latter is acid sugar peak.Through HPGPC, analyze, molecular weight 1.8060 ten thousand, the latter is acid sugar peak, through HPGPC, analyzes, Mw1.8687 ten thousand.
Embodiment 5
Get 80 of healthy rats, adaptability is raised after 3d, leaves and takes at random 10 as Normal group.Water 12h is can't help in all the other 70 rat fasting, by the dosage disposable celiac of 180mg/ (Kgbw), injects 2% tetraoxypyrimidine solution.After 72h, fasting 5h tail venous blood sampling is measured fasting plasma glucose, with blood glucose value 11.1～25mmol/L, and occurs that many drinks, many foods, diuresis person are defined as diabetes model.Select 50 of standard compliant animal patterns, by fasting plasma glucose and body weight level, be divided at random camellia polysaccharide (being obtained tea polysaccharide by embodiment 3) high dose group 800mg/ (kgbw), middle dosage group 400mg/ (kgbw), low dose group 200mg/ (kgbw), N1,N1-Dimethylbiguanide group 50mg/ (kgbw) and model control group, 10 every group.Each dosage group and N1,N1-Dimethylbiguanide group are given respectively corresponding medicine gavage, and normal group and model control group gavage physiological saline.Each group is all with the volume gavage of 1ml/100g.By 1 gavage of dose every day, continuous 15d.Test-results shows, before drug intervention, each group of injection tetraoxypyrimidine modeling is compared the fasting blood glucose level (P < 0.01) that all has high baseline with Normal group, and there was no significant difference (P > 0.05) between group.Experimental session diabetic model group shows lasting hyperglycemia.Blood sugar slow decreasing after each dosage group administration of camellia polysaccharide, tests while finishing and diabetic model group relatively has significant difference, especially with middle high dose group hypoglycemic activity obvious (P < 0.05).N1,N1-Dimethylbiguanide group is compared with diabetic model group utmost point significant difference (P < 0.01), the results are shown in Table 1.
The impact (X ± s) of table 1 camellia polysaccharide on diabetes rat the 1st day and the 15th day body weight and blood sugar
With normal group comparison, * * P < 0.01; With model control group comparison, △ P < 0.05, △ △ P < 0.01
Claims (3)
1. the application of camellia polysaccharide aspect preparation control diabetes medicament, is characterized in that, camellia polysaccharide is prepared by the following method:
A, by dry camellia after crushed, adds 5～20 times of mass ratioes of water or enzyme solution to extract after 30 minutes, filters or centrifugation obtains solid precipitation and extracting solution; Described being extracted at normal temperature to 100 ℃ carried out, and is to take the extraction process that water is medium, is one or more share of lixiviate, boiling water extraction, supersound extraction or microwave extraction;
B, concentrated extracting solution to density are greater than after 1.2Kg/L, add 2 times with upper volume lower alcohol, to staticly settle more than 12 hours, and supernatant liquor reclaims lower alcohol recycling, obtain camellia Crude polysaccharides after drying precipitate; Described lower alcohol is ethanol;
C, use macroporous adsorbent resin or ion exchange resin decolour and remove portion protein to camellia Crude polysaccharides, or use chemical deproteinated and decolouring to process; Or use filter membrane that molecular weight is less than to 10,000 camellia Crude polysaccharides, monose, inorganic salt and other Impurity removals and obtain camellia polysaccharide.
2. according to power, require camellia polysaccharide described in 1 in the application aspect preparation control diabetes medicament, it is characterized in that, in described water, add zymin, wherein the mass ratio of enzyme and water is 0.1～1.2:100.
3. according to power, require camellia polysaccharide described in 1 in the application aspect preparation control diabetes medicament, it is characterized in that, described enzyme comprises one or both zymin mixtures in polygalacturonase, cellulase.
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| CN102558379A (en) * | 2012-01-11 | 2012-07-11 | 新疆大学 | Method for extraction and deproteinization of hop polysaccharide |
| CN102948660B (en) * | 2012-10-11 | 2015-06-17 | 安徽双园茶业有限公司 | Trisaccharide crystal functional nutritional supplement |
| CN103059158A (en) * | 2013-01-29 | 2013-04-24 | 江西省蚕桑茶叶研究所 | Method for inhibiting foam generation in tea flower polysaccharide extraction |
| CN103497257B (en) * | 2013-09-27 | 2016-05-18 | 福建中烟工业有限责任公司 | A kind of Tea Flower polysaccharide, its preparation method and the application in cigarette thereof |
| CN104982465A (en) * | 2015-07-08 | 2015-10-21 | 杨君 | Compound for preventing white peony root leaf mould |
| CN106905412B (en) * | 2017-02-09 | 2020-08-07 | 上海师范大学 | Method for removing tea polysaccharide and glycoprotein in selenium-rich tea protein |
| CN107151275A (en) * | 2017-07-19 | 2017-09-12 | 上海家化联合股份有限公司 | Tea polysaccharide extract and its application in skin preparations for extenal use |
| CN107495370A (en) * | 2017-08-31 | 2017-12-22 | 湄潭县京贵茶树花产业发展有限公司 | The extracting method of active ingredient in Tea Flower |
| CN109078358A (en) * | 2018-08-23 | 2018-12-25 | 江苏天晟药业股份有限公司 | A kind of preparation method of bladder-wrack extract |
| CN111440252B (en) * | 2020-06-08 | 2021-09-21 | 湘丰茶业集团有限公司 | Method for extracting tea polysaccharide from tea leaves |
| CN115073623A (en) * | 2022-07-06 | 2022-09-20 | 湖北人人爱油脂有限公司 | Enzymatic preparation process of tea flower polyferose |
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