CN101658563B - Extraction method of raspberry for preventing primary carcinoma of liver - Google Patents
Extraction method of raspberry for preventing primary carcinoma of liver Download PDFInfo
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- CN101658563B CN101658563B CN2008101370049A CN200810137004A CN101658563B CN 101658563 B CN101658563 B CN 101658563B CN 2008101370049 A CN2008101370049 A CN 2008101370049A CN 200810137004 A CN200810137004 A CN 200810137004A CN 101658563 B CN101658563 B CN 101658563B
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Abstract
The invention discloses an extraction method of raspberry for preventing primary carcinoma of liver. The prior extraction method has many defects. The extraction method of the raspberry for preventing primary carcinoma of liver comprises: picking fresh raspberry in a ripening season, grinding the fresh raspberry and removing slag; fully stirring the ground raspberry from which the slag is removed, filling he ground raspberry in a breaker and placing the breaker on an ice block; adding 200 milliliters of 80 percent acetone in 100 grams of raspberry, and mixing the mixture in a pre-cooled mixer for 3 minutes; continuously homogenizing and stirring the mixture by using a homogenizer for 2 minutes; filling homogenate in a filter flask which is connected with a vacuum pump and provided with filter paper; collecting filtrate, filling the filtrate in a rotary evaporator and performing rotary evaporation at 50 DEG C for 45 minutes; adjusting the volume of the filtrate with deionized water to a final volume of 50 milliliters; and placing the solution at 20 DEG C below zero to 80 DEG C below zero for refrigeration. The method is an extraction method of raspberry.
Description
Technical field:
The present invention relates to a kind of extraction method of raspberry that prevents primary hepatocarcinoma.
Background technology:
Fructus Rubi (Raspberry) also claim rasp berry; Be Rosaceae rubus (Rubus sp.) plant; Contain defying age material such as a large amount of vitamin Cs, vitamin E, SOD (superoxide dismutase-content occupies first of the fruit), y-aminobutyric acid and cancer-resisting substance-tanning acid (Ellagic acid) in the fresh fruit, wherein vitamin E and tanning acid content be all fruit.The U.S. claims that red raspberry is the jinx of cancer, is described as " ruby ".Fructus Rubi is current fashionable America and Europe, the dark emerging fruit of high nutrition popular to people.Tanning acid has special efficacy to colon, cervix uteri, mammary gland and pancreatic cancer cell.Fructus Rubi and converted products thereof can keep the active component of tanning acid effectively, and can be absorbed by the body.Fructus Rubi is the current the highest food of finding of natural tanning acid content, and tanning acid is a kind of phenolic compound with phenyl ring, the growth of the sour cancerous cell that can also slow down of the tanning of low content, but high-load kill cancer cell then.
Present domestic existing scientific research group research Fructus Rubi, no matter but be on the direction of the purpose studied and research, very big-difference to be arranged all, very big difference is also arranged on extraction process.
The technology that Agricultural University Of Shenyang's Foodstuffs Academy extracts Fructus Rubi is to have adopted the method for SephadexG-100 column chromatography to carry out; This method has a lot of shortcomings: at first; Chromatography utilization positive and negative charge is inhaled principle mutually and is carried out; This method is lost neutral group base and weak charge group easily, and is well-known, and a lot of Organic substances all are neutral; Secondly, owing between the phytochemicals compatibility effect is arranged, chromatography method can not fully extract effective ingredient in the Fructus Rubi, so cause the unbalance of compatibility, the effect of phytochemicals can not be given full play to; Once more, chromatography needs the good fluidity of solution, causes the concentration of extract very low, is unfavorable for studying the effect of high concentration group.
College of science of Northeast Agricultural University should change system and study the technology that the hot-water process extracts the Fructus Rubi polysaccharide, and this method also has apparent in view inferior position: at first, the antioxidation composition is an Organic substance in the Fructus Rubi, and water can not fully extract it; Secondly, the antioxidation composition is oxidized as if there not being the effective measures meeting in hot water in the Fructus Rubi, thereby has lost the antioxidative ability, has not also just had the antineoplastic ability of raspberry extract.
Summary of the invention:
The purpose of this invention is to provide a kind of extraction method of raspberry that prevents primary hepatocarcinoma, can extract the effective ingredient in the Fructus Rubi fully and guarantee its antioxidation and the antineoplastic ability.
Above-mentioned purpose realizes through following technical scheme:
The extraction method of raspberry of prevention primary hepatocarcinoma; Its composition comprises: win fresh Fructus Rubi and smash in the mature period and remove slag, put into beaker after described Fructus Rubi of smashing after removing slag is fully stirred and place on the ice cube, in the 100g Fructus Rubi, add 80% acetone 200ml and in the blender of pre-cooling, mixed 3 minutes; Using homogenizer to continue homogenate then stirred 2 minutes; Take out homogenate and put into the filtration bottle that is connected to vacuum pump, filter to be placed with on the bottle and filter paper (Whatman2#), collect that permeate places rotary evaporator and rotary evaporation 45 minutes under 50 ℃ of conditions; Use deionized water to be adjusted to final volume and be 50ml, place subzero 20 ℃-subzero 80 ℃ frozen.
Beneficial effect of the present invention:
1. the present invention utilizes 80% acetone 200ml to mix with the 100g Fructus Rubi as solvent; This kind ratio is best proportioning; Can fully extract effective ingredient in the Fructus Rubi, the effect of phytochemicals is given full play to, and the concentration of extract is high; Can guarantee antioxidation composition antioxidative ability in the Fructus Rubi to greatest extent, thereby guarantee the antineoplastic ability of raspberry extract.Can filter more impurity with Whatman2# filter paper, make extract purer.
2. the fresh Fructus Rubi after the present invention will smash and remove slag is put into beaker and is placed on the ice cube, can reduce the color browning phytochemicals inactivation that causes owing to oxidation reaction.
3. through utilizing the TOSC method to detect the antioxidant activity of raspberry extract, the result shows that each experimental concentration group raspberry extract all has antioxidant activity, and along with the rising of concentration, antioxidant activity rises; Tumor prevented and treated to raspberry extract can from suppressing the kinds of tumors Expression of Related Genes; The anti-oxidation characteristics that phytochemicals in the raspberry extract has can be removed intravital oxygen-derived free radicals (ROS), the damage of prevention cell DNA, thereby the generation of prophylaxis of tumours; Total tanning acid (E llagic acid) can influence some BAs of tumor cell like propagation, oncogene expression, the generation of apoptosis etc.; Phytochemicals can suppress the expression of tumor cell VEGF; Influence the expression of wild type p53 and promote that the apoptosis of tumor cell possibly be the main mechanism of raspberry extract prevention and treatment malignant tumor; Experiment shows that raspberry extract can effectively bring into play the activity of anti-tumor in vivo in the body, and its effect is very obvious.
The specific embodiment:
Embodiment 1:
The extraction method of raspberry of prevention primary hepatocarcinoma; Its composition comprises: win fresh Fructus Rubi and smash in the mature period and remove slag, put into beaker after described Fructus Rubi of smashing after removing slag is fully stirred and place on the ice cube, in the 100g Fructus Rubi, add 80% acetone 200ml and in the blender of pre-cooling, mixed 3 minutes; Using homogenizer to continue homogenate then stirred 2 minutes; Take out homogenate and put into the filtration bottle that is connected to vacuum pump, filter and be placed with Whatman2# on the bottle and filter paper, collect that permeate places rotary evaporator and rotary evaporation 45 minutes under 50 ℃ of conditions; Use deionized water to be adjusted to final volume and be 50ml, place subzero 20 ℃-subzero 80 ℃ frozen.
The TOSC experiment is to utilize gas chromatography analysis method to detect the non-oxidizability of purification solution or biological sample.Under the condition that oxygen exists, ABAP can produce peroxide radical under hot conditions, and the further oxidation KMBA of these free radicals produces ethylene, and the output of ethylene can detect through gas chromatographic analysis.The theoretical basis of TOSC experiment is: antioxidant that adds and KMBA competition peroxide radical, thereby the partly generation of ethene suppressing.In our experiment response system; The raspberry extract of variable concentrations (200 μ g/ml, 400 μ g/ml, 800 μ g/ml, 1000 μ g/ml) adds in the Teflon/Silicone bottle of 4ml band Jiao Gai; Contain 0.2mM KMBA in the bottle; 20mM ABAP and each 0.1ml of 100mM phosphate buffer use deionized water to regulate final volume at last and are 1ml.Entire reaction is activated behind the 200mM ABAP adding 100 μ l.At 37 ℃, the shaking table internal reaction under the 160r/min condition 15,30,45,60 minutes, matched group contain other all reagent except that raspberry extract.Gas above the different periods see through bottle bottle stopper extraction 0.5ml reactant liquor is in the injection gas chromatography response system.Helium is as transporting gas, and flow velocity is 25ml/min.
Through utilizing the TOSC method to detect the antioxidant activity of raspberry extract; The result shows that each experimental concentration group raspberry extract all has antioxidant activity; Along with the rising of concentration, antioxidant activity rises, and the mechanism of the removing free radical that has been found that at present has two kinds: a kind of is to strengthen activities of antioxidant enzymes; Eliminating rapidly free radical, is exactly the important enzyme system that is present in the removing oxygen-derived free radicals in the middle of the human normal tissue such as superoxide dismutase and catalase; A kind of in addition method is directly to take in antioxidant.It is reported: the main antioxidant in vegetable and the fruit is Polyphenols and flavone compound, and the ascorbic non-oxidizability that derives from the Fructus Mali pumilae only accounts for 0.4% of total oxidation resistance.In the Fructus Rubi phytochemicals antioxidant activity of tanning acid (Ellagicacid) be Vc 600-800 doubly, explain that the stronger antioxidant activity that Fructus Rubi has is mainly derived from other plant chemical ingredient, they are one of the main reasons of its prophylaxis of tumours generation.
The mechanism of cancer is very complicated, not only relates to the variation of multiple oncogene, antioncogene, and some cytokines also play a role in the process of cancer generation, development.Although the model of action of these genes, cytokine pair cell is different, action pathway and site of action are different, and final result all will cause the malignant proliferation of cell.Therefore, the malignant proliferation of containment cell is the basis of oncotherapy.Our experimental result shows that raspberry extract raises with concentration, can obviously suppress tumor cell proliferation, thus the generation of prophylaxis of tumours.
In human tumor, the sudden change of antioncogene p53 is one of more common reason, and is more obvious with performance in the colorectal cancer especially.The p53 gene has two kinds of wild type and saltants, and the former is a tumor suppressor gene, and the latter is an oncogene, and the generation that all can promote tumor is expressed in crossing of the former sudden change, disappearance or the latter.The wild type p53 albumen that our experiment detects, its positive expression has cancer suppressing action, and experiment shows that the phytochemicals in the raspberry extract can strengthen the expression of wild type p53.
Numerous researchs show that VEGF has highly expression in nearly all malignant tumor tissue, and higher vegf expression rate has reflected high-caliber proliferative activity.Result of study shows that raspberry extract has the activity of obvious suppression vegf expression, makes tumor cell be in lower vegetative state, and suppresses the generation and the development of tumor.The proteic expression excessively of VEGF high expressed and wild p53 is negative correlation, and the incidence and development of prompting cancerous protuberance is relevant with kinds of tumors related gene synergism, and tumor prevented and treated to raspberry extract can from suppressing the kinds of tumors Expression of Related Genes.
In a word, the anti-oxidation characteristics that the phytochemicals in the raspberry extract has can be removed intravital oxygen-derived free radicals (ROS), the damage of prevention cell DNA, thereby the generation of prophylaxis of tumours; Total tanning acid (E llagic acid) can influence some BAs of tumor cell like propagation, oncogene expression, the generation of apoptosis etc.; Phytochemicals can suppress the expression of tumor cell VEGF; Influence the expression of wild type p53 and promote that the apoptosis of tumor cell possibly be the main mechanism of raspberry extract prevention and treatment malignant tumor; Experiment shows that raspberry extract can effectively bring into play the activity of anti-tumor in vivo in the body, and its effect is very obvious.
At present the sickness rate of malignant tumor is the trend that rises year by year, but treatment means mainly is to be main with operation, and chemotherapy, radiotherapy are auxilliary, and curative effect is not good enough, and mortality rate is in first of all kinds of diseases.Therefore, prevention, the treatment problem of relevant malignant tumor are the matters of utmost importance that contemporary medical personnel faces.With regard to Drug therapy, most anticarcinogens of using clinically at present are mainly chemical synthetic drug, and its mechanism of action is main to suppress and to kill tumor cell mainly, and often causes the damage of normal tissue cell, so toxicity, side effect is bigger.In addition, malignant cell is easy to generate drug resistance after contacting medicine for a long time, has influenced therapeutic effect widely.Natural plant extracts, its various compositions are natural compatibility, almost do not have cytotoxicity and side effect, can be used as long-term prevention and medicine for treatment.Raspberry extract has very strong antioxidant activity; Activity with tangible anti-SMMC-7721 HepG-2 propagation; Raspberry extract can suppress expression and the inducing tumor cell generation apoptosis of SMMC-7721 HepG-2VEGF; Raspberry extract can influence the expression of the wild p53 of SMMC-7721 HepG-2; Raspberry extract suppresses VEGF and influences the expression of wild p53 and three main aspects that inducing apoptosis of tumour cell is its antitumor mechanism; Raspberry extract can obviously suppress the generation and the progress of tumor in vivo.
Claims (1)
1. extraction method of raspberry that prevents primary hepatocarcinoma; Its composition is: win fresh Fructus Rubi and smash in the mature period and remove slag; It is characterized in that: put into beaker after described Fructus Rubi of smashing after removing slag is fully stirred and place on the ice cube; The acetone 200ml of adding 80% mixed 3 minutes in the blender of pre-cooling in the 100g Fructus Rubi, used homogenizer to continue homogenate then and stirred 2 minutes, took out homogenate and put into the filtration bottle that is connected to vacuum pump; Filter on the bottle and be placed with filtration paper; Collect that permeate places rotary evaporator and under 50 ℃ of conditions rotary evaporation 45 minutes, use deionized water to be adjusted to final volume and be 50ml, place subzero 20 ℃-subzero 80 ℃ frozen.
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| CN104758270B (en) * | 2015-04-04 | 2018-07-06 | 青海瑶池生物科技有限公司 | A kind of raspberry polysaccharide effervescence tablet and application thereof |
| CN104873532B (en) * | 2015-04-04 | 2018-06-19 | 中国科学院西北高原生物研究所 | Application of the raspberry polysaccharide in antitumor |
| CN105687422B (en) * | 2016-03-02 | 2020-05-05 | 深圳福山生物科技有限公司 | Raspberry composition and application thereof |
Citations (2)
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| CN101129152A (en) * | 2007-10-10 | 2008-02-27 | 辽宁今日农业有限公司 | Method of preparing blueberry leaf tea beverage |
| CN101133771A (en) * | 2007-10-10 | 2008-03-05 | 辽宁今日农业有限公司 | Method of producing bramble leaf tea beverage |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101129152A (en) * | 2007-10-10 | 2008-02-27 | 辽宁今日农业有限公司 | Method of preparing blueberry leaf tea beverage |
| CN101133771A (en) * | 2007-10-10 | 2008-03-05 | 辽宁今日农业有限公司 | Method of producing bramble leaf tea beverage |
Non-Patent Citations (4)
| Title |
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| 徐俐等.树莓红色素的提取及稳定性的研究.《食品科学》.2006,第27卷(第05期),191-194. * |
| 李晓燕等.树莓色素稳定性初步研究.《饮料工业》.2005,第8卷(第02期),11-13. * |
| 王象欣等.树莓中多糖提取工艺的研究.《食品科技》.2007,(第05期),107-109. * |
| 韩泽梅等.树莓种子原花青素的提取分离工艺研究.《天然产物研究与开发》.2006,第18卷(第01期),108-111. * |
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