CN102399299A - Preparation method for tuckahoe acidic polysaccharide extract and application of tuckahoe acidic polysaccharide extract - Google Patents
Preparation method for tuckahoe acidic polysaccharide extract and application of tuckahoe acidic polysaccharide extract Download PDFInfo
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Abstract
The invention belongs to the technical field of extraction and purification of active parts of Chinese medicines and in particular discloses a preparation method for a tuckahoe acidic polysaccharide extract. The preparation method comprises the following steps of: soaking tuckahoe into 60 to 80 v/v percent ethanol in amount which is 4 to 8 times the weight of the tuckahoe for 3 to 12 hours; performing refluxing extraction for 1 to 2 hours at each time, totally for 1 to 3 times; filtering to obtain filter residues; performing refluxing extraction on the filter residues by using water in amount which is 4 to 10 times the weight of the filter residues for 1 to 3 hours at each time, totally for 1 to 3 times; filtering to obtain filter residues; extracting by adding 0.1 to 0.5 M NaOH in an amount which is 30 to 70 times the weight of the filter residues for 0.5 to 2 hours; filtering; neutralizing the filtrate by using HCl; washing and desalting to obtain precipitate; and performing spray drying on the precipitate to obtain the tuckahoe acidic polysaccharide extract. The method is simple; the used solvents can be reused; production cost is reduce; energy is saved; the Tuckahoe acidic polysaccharide content of the obtained extract is 1.9 to 50.0 weight percent based on the weight of glucose. The extract has the functions of obviously regulating immunity and resisting tumor.
Description
Technical field
The preparation method and the application thereof of pharmaceutically active ingredient in the present invention relates to more specifically relate to the preparation method and the application in the preparation medicament for immunity enhancement thereof of the acid polyoses extract of a kind of Poria cocos.
Background technology
Poria cocos Poria cocos (Schw.) Wolf is conventional Chinese medicine, medicine-food two-purpose since ancient times.History is stated from Shennong's Herbal, and quilt is classified as top grade, is the dry sclerotia of polyporaceae fungi Poria cocos (Schw.) Wolf.The property slightly sweet flavor, flat, be included into heart spleen lung channel.Effect with clearing damp Li Shui, invigorating the spleen and regulating the stomach, tranquillizing by calming the heart.Clinically be used to treat dysuria, the oedema turgor, phlegm and retained fluid is coughed contrary, and it is dirty to vomit, have loose bowels, seminal emission, stranguria with turbid discharge, palpitation with fear, forgetful etc.The major ingredient of Poria cocos is Pachymose, triterpene compound, protein, fat and inorganic salt etc.Wherein Pachymose and triterpene are its main active ingredient, and Chinese scholars has been done more research to the pharmacological action and the clinical application aspect of compositions such as Pachymose, triterpene, and bigger progress has been arranged.
Research shows that Pachymose has antitumor action, can increase gerontal patient's humoral immune function, reduces the spinoff that produces in the white blood disease radiotherapy.
Acidic polysaccharose content accounts in the Poria cocos medicinal material more than 50% in the Poria cocos; Be water insoluble active ingredient; But the methods that adopt water boiling and extraction in the clinical application more; What pharmaceutical research also was primarily aimed at is water-soluble portion in the Poria cocos, very few to its caustic solubility part Study, thereby has caused loss of active ingredients in the Poria cocos.In conjunction with modern chemistry and pharmacological research achievement, the Pachymose constituents is developed, excavate new drug, will realistic meaning be arranged to the exploitation and the application of traditional Chinese medicine Poria cocos.
Summary of the invention
An object of the present invention is to provide the preparation method of the acid polyoses extract of a kind of Poria cocos.
The acid polyoses extract of Poria cocos that another object of the present invention has provided a kind of method for preparing has the application in the medicine of immunoregulation effect in preparation.
In order to realize above-mentioned first purpose, the technical measures that the present invention adopts are following:
The preparation method of the acid polyoses extract of a kind of Poria cocos, its step is following:
(1) take by weighing Poria cocos, extraordinarily going into concentration according to the 4-8 of its weight is alcohol immersion 3-12 hour of 60～80v/v%;
(2) soak the back refluxing extraction 1-3 time, each time is 1-2 hour, filters, and abandons extracting solution, gets filter residue;
Accomplish the back direct heating in the immersion of step (1) and carry out refluxing extraction first time, isolate extracting solution after having extracted, the ethanol of the 70v/v% of 8 times of raw material Poria cocos weight of adding is as extraction agent during follow-up each extraction;
(3) filter residue of step (2) is added water logging bubble 3-12 hour of 4-10 times of weight, soak the back refluxing extraction 1-3 time, each time is 1-3 hour, filters, and abandons extracting solution, gets filter residue;
Accomplish the back direct heating in the immersion of step (3) and carry out the refluxing extraction first time, isolate extracting solution after having extracted, the water of filter residue weight that adds 4-10 times of step (2) during follow-up each extraction again is as extraction agent;
(4) filter residue that step (3) is obtained pulverizes, and adds its weight 30-70 0.1-0.5mol/L NaOH solution stirring doubly and extracts 30-120min, filters, and filtrating adds 2wt%HCl and is neutralized to neutrality, and spinning goes out deposition, uses water washing and precipitating with desalination again, must precipitate;
(5) step (4) gained deposition is spray-dried, promptly get the acid polyoses extract product of Poria cocos.
In step (1) (2), the purpose of alcohol reflux is to isolate the Poria cocos triterpenes components.
In step (3), the purpose of water refluxing extraction is to isolate the Poria cocos water-soluble polysaccharide.
In step (4), the optimum concn of NaOH is 0.5mol/L, and its optimum amount is 70 times of the filter residue weight that obtains of step (3).
In step (5), described spraying drying operational condition is following: intake air temperature is 175 ℃, and the air outlet temperature is 90 ℃, and the atomizing disk rotating speed is 29040 rev/mins.
Polysaccharide content is 1.9-50.0wt% with the glucose meter in the acid polyoses extract of the Poria cocos that utilizes above-mentioned preparation method to obtain.
The acid polyoses extract of the Poria cocos that method for preparing obtains can be used for preparing medicament for immunity enhancement.
Compared with prior art, advantage of the present invention and beneficial effect are:
The present invention has improved the utilization ratio of Poria cocos medicinal material, has opened up its new medicinal approach.
The effect that the acid polyoses extract of the Poria cocos of gained of the present invention has good adjusting immunologic function.Not only can use separately and can also unite use with the other drug compatibility.
Embodiment
Further set forth method of the present invention and application thereof below in conjunction with concrete embodiment.
Poria cocos is a kind of conventional Chinese medicine material, generally uses with the form of medicine materical crude slice, and this medicinal material can directly be buied from each Poria cocos place of production, and the Poria cocos sheet that uses in the inventive method is available from the nine Zi He towns, Luotian County, Hubei Province of the title that " township of Poria cocos " arranged.
Embodiment 1:
The preparation method of the acid polyoses extract of a kind of Poria cocos, step is following:
(1) takes by weighing the Poria cocos sheet, extraordinarily go into the alcohol immersion 12 hours that concentration is 70v/v% according to 8 of its weight;
(2) soaking the back refluxing extraction 2 times, is 2h for the first time, for 1h (the 70v/v% ethanol that adds 8 times of Poria cocos sheet weight when extracting for the second time), filters for the second time, obtains filter residue;
(3) 12h is steeped in the water logging that adds 8 times of steps (2) filter residue weight;
(4) soak the back refluxing extraction 3 times, extract 1h at every turn, back extracted twice water consumption is followed successively by 6 times and 4 times of step (2) filter residue weight; Filter, obtain filter residue;
(5) filter residue is pulverized, the NaOH that adds the 0.5mol/L of 70 times of steps (4) gained filter residue weight stirs and extracts 1h; Filter, must filtrate;
(6) in above-mentioned filtrating, add 2wt%HCl and be neutralized to neutrality, the centrifugal acid polysaccharide crude of Poria cocos that obtains;
(7) with three washings of the acid polysaccharide crude of above-mentioned Poria cocos centrifugal after, must precipitate, again gained is precipitated spraying drying, promptly get Poria cocos acidity polyoses extract.
Through test, in the acid polyoses extract of the Poria cocos that makes, the content of polysaccharide is 21.8wt% (with glucose meter), and dried cream yield is 24.5% (weight).
Embodiment 2:
The preparation method of the acid polyoses extract of a kind of Poria cocos, step is following:
(1) takes by weighing the Poria cocos sheet, extraordinarily go into the alcohol immersion 10 hours that concentration is 80v/v% according to 4 of its weight;
(2) soaking the back refluxing extraction 2 times, is 2h for the first time, for 1h (the 70v/v% ethanol that adds 8 times of Poria cocos sheet weight when extracting for the second time), filters for the second time, obtains filter residue;
(3) 8h is steeped in the water logging that adds 10 times of steps (2) filter residue weight;
(4) soak the back refluxing extraction 3 times, extract 1h at every turn, back extracted twice water consumption is followed successively by 8 times and 6 times of step (2) filter residue weight; Filter, obtain filter residue;
(5) filter residue is pulverized, the NaOH that adds the 0.2mol/L of 70 times of steps (4) gained filter residue weight stirs and extracts 1h; Filter, must filtrate;
(6) in above-mentioned filtrating, add 2wt%HCl and be neutralized to neutrality, the centrifugal acid polysaccharide crude of Poria cocos that obtains;
(7) with four washings of the acid polysaccharide crude of above-mentioned Poria cocos centrifugal after, must precipitate, again gained is precipitated spraying drying, promptly get Poria cocos acidity polyoses extract.
Through test, in the acid polyoses extract of the Poria cocos that makes, the content of polysaccharide is 1.7wt% (with glucose meter), and dried cream yield is 9.8% (weight).
Embodiment 3:
The preparation method of the acid polyoses extract of a kind of Poria cocos, step is following:
(1) takes by weighing the Poria cocos sheet, extraordinarily go into the alcohol immersion 6 hours that concentration is 70v/v% according to 7 of its weight;
(2) soaking the back refluxing extraction 2 times, is 2h for the first time, for 1h (the 70v/v% ethanol that adds 8 times of Poria cocos sheet weight when extracting for the second time), filters for the second time, obtains filter residue;
(3) 6h is steeped in the water logging that adds 8 times of steps (2) filter residue weight;
(4) soak the back refluxing extraction 3 times, extract 1h at every turn, back extracted twice water consumption is followed successively by 4 times and 6 times of step (2) filter residue weight; Filter, obtain filter residue;
(5) filter residue is pulverized, the NaOH that adds the 0.2mol/L of 70 times of steps (4) gained filter residue weight stirs and extracts 2h; Filter, must filtrate;
(6) in above-mentioned filtrating, add 2wt%HCl and be neutralized to neutrality, the centrifugal acid polysaccharide crude of Poria cocos that obtains;
(7) with five washings of the acid polysaccharide crude of above-mentioned Poria cocos centrifugal after, must precipitate, again gained is precipitated spraying drying, promptly get Poria cocos acidity polyoses extract.
Through test, in the acid polyoses extract of the Poria cocos that makes, the content of polysaccharide is 1.69wt% (with glucose meter), and dried cream yield is 9.8% (weight).
Embodiment 4:
The preparation method of the acid polyoses extract of a kind of Poria cocos, step is following:
(1) takes by weighing the Poria cocos sheet, extraordinarily go into the alcohol immersion 3 hours that concentration is 60v/v% according to 6 of its weight;
(2) soaking the back refluxing extraction 2 times, is 2h for the first time, for 1h (the 70v/v% ethanol that adds 8 times of Poria cocos sheet weight when extracting for the second time), filters for the second time, obtains filter residue;
(3) 3h is steeped in the water logging that adds 8 times of steps (2) filter residue weight;
(4) soak the back refluxing extraction 3 times, extract 1h at every turn, back extracted twice water consumption is followed successively by 4 times and 6 times of step (2) filter residue weight; Filter, obtain filter residue;
(5) filter residue is pulverized, the NaOH that adds the 0.5mol/L of 70 times of steps (4) gained filter residue weight stirs and extracts 1.5h; Filter, must filtrate;
(6) in above-mentioned filtrating, add 2wt%HCl and be neutralized to neutrality, the centrifugal acid polysaccharide crude of Poria cocos that obtains;
(7) with three washings of the acid polysaccharide crude of above-mentioned Poria cocos centrifugal after, must precipitate, again gained is precipitated spraying drying, promptly get Poria cocos acidity polyoses extract.
Through test, in the acid polyoses extract of the Poria cocos that makes, the content of polysaccharide is 23.6wt% (with glucose meter), and dried cream yield is 25.0% (weight).
Embodiment 5: the pilot scale amplification test
The preparation method of the acid polyoses extract of a kind of Poria cocos:
Take by weighing Poria cocos sheet 20kg, the 70v/v% alcohol immersion that adds its 8 times of weight is after 12 hours, refluxing extraction twice, and 2h for the first time adds for the second time the 70v/v% alcohol reflux 1h of 8 times of raw material Poria cocos sheet weight, filters; The water logging bubble 12h that in filter residue, adds its 8 times of weight, refluxing extraction 3 times (during back extracted twice amount of water with for the first time with), each 1h filters, and gets filter residue; Filter residue is pulverized, and the 0.5mol/LNaOH that adds 70 times of filter residue weight stirs and extracts 1h, filters; Filtrating adds 2wt%HCl and is neutralized to neutrality, the acid polysaccharide crude of the centrifugal Poria cocos that obtains after washing desalination three times, spraying drying again; Collect dry thing, promptly get the acid polyoses extract pilot product of Poria cocos, sealing; Weigh, place dry place to preserve.Three crowdes of pilot scale amplification test results see table 1.
Three batches of pilot scale scale-ups of table 1 are table as a result
The result shows: the processing parameter of the present invention's research is feasible, can carry out the transition to industrialization production.
Embodiment 6:
The acid polyoses extract of Poria cocos for preparing with the foregoing description 1 carries out pharmacological evaluation, pharmacology result and analysis, and the acid polyoses extract of Poria cocos that makes with proof the inventive method can be applied to prepare in the medicine with immunoregulation effect.
The experimental study of the immunoregulation effect of the acid polyoses extract of Poria cocos:
Kunming mice (18-22g), male and female half and half, random packet, 10 every group.Gastric infusion 14 days, preceding 4 days blank control group and model group give saline water 10ml/kg/d, and other two groups give the acid polyoses extract of dried Poria cocos and (divide height two dose groups; Consumption is respectively: 100mg/kg/d, 300mg/kg/d), from the 5th day by the 10th day, except that blank control group; (specification of hydrocortisone injection: 5mg/ml), stopped modeling in back 4 days, blank control group and model group give saline water 10ml/kg/d to other each group difference subcutaneous injection HYDROCORTISONE INJECTIONS 10ml/kg/d; Other two groups give the acid polyoses extract of dried Poria cocos (consumption still are respectively 100mg/kg/d; 300mg/kg/d), pluck eyeball and get blood, separation of serum next day after the last administration; Press the content that test kit specification sheets method is measured interleukin II (IL-2), the bad factor of tumour (TNF α) and IFN-r, the result sees table 2; Get thymus gland, spleen mensuration organ index (organ weights/body weight * 10), the result sees table 3:
The acid polyoses extract of table 2 Poria cocos is to the influence
of mice serum IL-2 and TNF alpha content
Annotate: compare P＜0.05 with saline water+HYDROCORTISONE INJECTIONS group (model group)
*, P＜0.01
△
The acid polyoses extract of table 3. Poria cocos is to the influence
of mouse thymus, index and spleen index
Annotate: compare P＜0.05 with saline water+HYDROCORTISONE INJECTIONS group (model group)
*.
Above experimental result fully shows, compares with model group, and the acid polyoses extract of Poria cocos can obviously improve by the immunocompromised immune function of mice due to the HYDROCORTISONE INJECTIONS, can obviously improve thymus index and the index and spleen index of mouse; And can obviously promote cytokines such as T cell synthetic more IL-2, TNF-α and IFN-r, thereby improve the immunological competence of body.
Claims (7)
1. the preparation method of the acid polyoses extract of a Poria cocos, step is following:
(1) take by weighing Poria cocos, extraordinarily going into concentration according to the 4-8 of its weight is alcohol immersion 3-12 hour of 60-80v/v%;
(2) soak the back refluxing extraction 1-3 time, each time is 1-2 hour, filters, and gets filter residue;
Accomplish the back direct heating in the immersion of step (1) and carry out refluxing extraction first time, isolate extracting solution after having extracted, the ethanol of the 70v/v% of 8 times of raw material Poria cocos weight of adding is as extraction agent during follow-up each extraction;
(3) the 4-10 water logging doubly that in the filter residue of step (2), adds its weight was steeped 3-12 hour, soaked the back refluxing extraction 1-3 time, and each extraction time is 1-3 hour, filters, and gets filter residue;
Accomplish the back direct heating in the immersion of step (3) and carry out the refluxing extraction first time, isolate extracting solution after having extracted, the water of filter residue weight that adds 4-10 times of step (2) during follow-up each extraction again is as extraction agent;
(4) filter residue that step (3) is obtained pulverizes, and the 30-70 0.1-0.5mol/L NaOH solution stirring doubly that adds its weight is extracted 30-120min, filters; Filtrating adds 2wt%HCl and is neutralized to neutrality; Spinning goes out deposition, uses water washing and precipitating with desalination again, must precipitate;
(5) gained is precipitated spray-dried, promptly get the acid polyoses extract product of Poria cocos.
2. preparation method according to claim 1 is characterized in that: described spraying drying operational condition is following: intake air temperature is 175 ℃, and the air outlet temperature is 90 ℃, and the atomizing disk rotating speed is 29040 rev/mins.
3. preparation method according to claim 1 and 2 is characterized in that: the alcohol concn described in the step (1) is 70v/v%, and amount of ethanol is 8 times of Poria cocos weight.
4. preparation method according to claim 1 and 2 is characterized in that: the refluxing extraction number of times described in the step (2) is 2 times, and the time is 2 hours for the first time, and the time is 1 hour for the first time.
5. preparation method according to claim 1 and 2 is characterized in that: the each extraction time described in the step (3) is 1 hour, and the number of times of refluxing extraction is 3 times.
6. preparation method according to claim 1 and 2 is characterized in that: the concentration of the NaOH solution described in the step (4) is 0.5mol/L, and consumption is 70 times of the filter residue weight that obtains of step (3), and extraction time is 1 hour.
7. the acid polyoses extract of the Poria cocos that makes according to arbitrary described preparation method among the claim 1-6 has the application in the medicine of antitumor and immunoregulation effect in preparation.
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| CN109748981A (en) * | 2017-11-02 | 2019-05-14 | 中国科学院微生物研究所 | A kind of alkali carries method and its application of pachymaran |
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| CN103451022A (en) * | 2013-09-02 | 2013-12-18 | 南昌大学 | Method for integrally extracting volatile oil, polysaccharide and flavone from elsholtzia haichowensis sun |
| CN103451022B (en) * | 2013-09-02 | 2014-12-31 | 南昌大学 | Method for integrally extracting volatile oil, polysaccharide and flavone from elsholtzia haichowensis sun |
| WO2016062020A1 (en) * | 2014-10-22 | 2016-04-28 | 北京中安佐际生物科技有限公司 | Pachyman active components and ingredients, preparation method therefor and use thereof |
| CN104666369A (en) * | 2015-03-16 | 2015-06-03 | 湖南省中医药研究院 | Extraction method for poria peel |
| CN104666369B (en) * | 2015-03-16 | 2018-04-27 | 湖南省中医药研究院 | A kind of extracting method of fuling peel |
| CN106866831A (en) * | 2017-01-22 | 2017-06-20 | 嵊州市派特普科技开发有限公司 | The Preparation method and use of Poria cocos acidity polyoses extract |
| CN109748981A (en) * | 2017-11-02 | 2019-05-14 | 中国科学院微生物研究所 | A kind of alkali carries method and its application of pachymaran |
| CN107814826A (en) * | 2017-11-06 | 2018-03-20 | 长沙爱扬医药科技有限公司 | A kind of method that pachymic acid and pachymaran are extracted from fuling peel |
| CN109988248A (en) * | 2017-12-29 | 2019-07-09 | 瑞普（天津）生物药业有限公司 | A kind of preparation method and application of Poria cocos acidic polysaccharose |
| US20220016196A1 (en) * | 2020-07-09 | 2022-01-20 | Unigen, Inc. | Aloe Based Compositions Comprising Polysaccharides and Polyphenols for Regulation of Homeostasis of Immunity |
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Inventor after: Zhang Weiyuan Inventor after: Liu Yanwen Inventor after: Liu Guoqing Inventor after: Wang Guangzhong Inventor after: Ye Xiaochuan Inventor after: Deng Yuanyuan Inventor after: Shao Beibei Inventor after: Zhang Yongdan Inventor before: Zhang Weiyuan Inventor before: Liu Yanwen Inventor before: Liu Guoqing Inventor before: Wang Guangzhong Inventor before: Ye Xiaochuan Inventor before: Deng Aiai Inventor before: Shao Beibei Inventor before: Zhang Yongdan |
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Address after: 432400, Hubei should be in the south of the City Economic Development Zone, all the way Patentee after: Hubei Hui Sheng Biotechnology Co., Ltd. Address before: 432400, Hubei should be in the south of the City Economic Development Zone, all the way Patentee before: Hubei Yangsheng Biotechnology Co., Ltd. |