CN104892789A - Celandine polysaccharide extracted from Chelidonium majus and application of celandine polysaccharide - Google Patents
Celandine polysaccharide extracted from Chelidonium majus and application of celandine polysaccharide Download PDFInfo
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Abstract
The invention discloses celandine polysaccharide extracted from Chelidonium majus and application of celandine polysaccharide and aims to achieve effectiveness in extraction of celandine polysaccharide and applicability to medicines for improving immunity. The technical scheme includes that adding water to Chelidonium majus powder, extracting, filtering and carrying out vacuum concentration to obtain polysaccharide concentrated solution; adding ethanol for precipitation, washing to obtain crude polysaccharide, sampling resin columns, subjecting eluent to vacuum concentration, adding ethanol for precipitation and washing to obtain celandine polysaccharide. In a process of extraction of the celandine polysaccharide from Chelidonium majus, raw materials which are natural traditional Chinese medicinal materials are extensive in source and low in cost. The celandine polysaccharide is taupe powder and has an excellent function for improving body immunity, application of Chelidonium majus to preparation of healthcare foods and medicines for improvement of body immunity creates a new application prospect of Chelidonium majus, and high social benefits and economic benefits are achieved while innovations on traditional Chinese medicine are realized.
Description
Technical field
The present invention relates to field of medicaments, particularly relate to a kind of pilewort polysaccharide of extracting from pilewort and application thereof.
Background technology
In recent years, along with the progress of society's science and technology, the enhancing of growth in the living standard and public's self health consciousness, sight focuses on and can strengthen on the product of immunization by increasing people, greatly facilitates the research of immunostimulant product.Vegetable polysaccharides is the composition that a class is extracted from natural drug, and modern study shows, vegetable polysaccharides has the activity improving the aspect such as body immunity, antitumor, antiviral, anti-oxidant, anti-ageing, hypoglycemic, reducing blood-fat, more ulcer.Further, much research shows, polysaccharide has stronger biological activity, is the principle active component of natural drug performance effect.
Pilewort is the dry herb of papaveracease Chelidonium plant pilewort (Chelidonium majus L), is the newly-increased kinds of 2010 editions Pharmacopoeias of People's Republic of China ().Its bitter, cool in nature, poisonous: all herbal medicine, there are spasmolysis and analgesia, relieving cough and asthma effect; For gastral cavilty contraction pain, cough and asthma, Whooping cough.The research of current Natural Medicine Chemistry and pharmacology aspect mainly concentrates on the alkaloids composition aspect of pilewort, but has no report to the extraction and isolation of its polyose composition and the research of pharmacologically active thereof.
Summary of the invention
For above-mentioned situation, for overcoming the defect of prior art, the object of the present invention is just to provide a kind of pilewort polysaccharide of extracting from pilewort and application thereof, effectively can solve the extraction of pilewort polysaccharide, and be effective to the problem in strengthening immunity product.
The technical scheme that the present invention solves is, a kind of pilewort polysaccharide extracted from pilewort, and the extracting method of described pilewort polysaccharide comprises the following steps:
(1) get pilewort medicinal powder, add the water of 10 ~ 40 times of pilewort medicinal powder weight, 60 ~ 100 DEG C are extracted 2 ~ 3 times, each 1 ~ 3h, and filter, merging filtrate, is evaporated to 40 DEG C of relative densities 1.10 ~ 1.20, obtains polysaccharide concentrated solution;
(2) in the polysaccharide concentrated solution of step (1), add volume fraction 95% ~ 100% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 50 ~ 100 times, acetone, anhydrous diethyl ether, 1 ~ 2 time is respectively washed successively to gained throw out, drying, obtains Crude polysaccharides;
(3) the Crude polysaccharides 0.2 ~ 1.0g getting step (2) is dissolved in 10mL water, be splined on HPD-600 type macroporous adsorptive resins, and with 200 ~ 300mL water elution, flow velocity is 0.5 ~ 1.5mL/min, collect elutriant, then be evaporated to the 1/2-2/3 of elutriant original volume; Add volume fraction 95% ~ 100% ethanol and reach 80% to alcohol content, hold over night, gets bottom settlings, respectively washs 1 ~ 2 time successively to gained throw out with the dehydrated alcohol of weight of precipitate 50 ~ 100 times, acetone, anhydrous diethyl ether, drying, obtains pilewort polysaccharide.
Pilewort medicinal powder in step (1) was the powder of No. 2 pharmacopeia sieves.
Technical program of the present invention also lies in providing a kind of pilewort polysaccharide to strengthen the application in immunologic function medicine in preparation.
Technical program of the present invention also lies in providing a kind of pilewort polysaccharide to strengthen the application in immunologic function protective foods in preparation.
The present invention extracts pilewort polysaccharide from pilewort, and raw material is natural traditional Chinese medicine, and wide material sources are with low cost.Pilewort polysaccharide of the present invention is grey-brown powder, there is the effect of stronger enhancing body immunizing power, be applied in medicine, the application of pilewort in the medicine and protective foods of preparation enhancing body immunizing power can be realized, open the new prospect of application of pilewort medicinal material, having good Social benefit and economic benefit, is the innovation on Chinese medicine.Meanwhile, the inventive method is simple, is easy to industrialization promotion.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is elaborated.
Embodiment 1
The extracting method of the pilewort polysaccharide of the present embodiment comprises the following steps:
(1) get pilewort medicinal powder, add the water of 10 times of pilewort medicinal powder weight, 60 DEG C are extracted 2 times, each 1h, and filter, merging filtrate, is evaporated to 40 DEG C of relative densities 1.10, obtains polysaccharide concentrated solution;
(2) in the polysaccharide concentrated solution of step (1), add volume fraction 95% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether, 1 time is respectively washed successively to gained throw out, drying, obtains Crude polysaccharides;
(3) the Crude polysaccharides 0.2g getting step (2) is dissolved in 10mL water, is splined on HPD-600 type macroporous adsorptive resins, and with 200mL water elution, flow velocity is 0.5mL/min, collects elutriant, then is evaporated to 1/2 of elutriant original volume; Add volume fraction 95% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether, 1 time is respectively washed successively to gained throw out, dry, obtain pilewort polysaccharide.
Embodiment 2
The extracting method of the pilewort polysaccharide of the present embodiment comprises the following steps:
(1) get pilewort medicinal powder, add the water of 40 times of pilewort medicinal powder weight, 100 DEG C are extracted 3 times, each 3h, and filter, merging filtrate, is evaporated to 40 DEG C of relative densities 1.20, obtains polysaccharide concentrated solution;
(2) in the polysaccharide concentrated solution of step (1), add volume fraction 100% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 100 times, acetone, anhydrous diethyl ether, 2 times are respectively washed successively to gained throw out, drying, obtains Crude polysaccharides;
(3) the Crude polysaccharides 1.0g getting step (2) is dissolved in 10mL water, is splined on HPD-600 type macroporous adsorptive resins, and with 300mL water elution, flow velocity is 1.5mL/min, collects elutriant, then is evaporated to 2/3 of elutriant original volume; Add volume fraction 100% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 100 times, acetone, anhydrous diethyl ether, 2 times are respectively washed successively to gained throw out, dry, obtain pilewort polysaccharide.
Embodiment 3
The extracting method of the pilewort polysaccharide of the present embodiment comprises the following steps:
(1) get pilewort medicinal powder, add the water of 25 times of pilewort medicinal powder weight, 80 DEG C are extracted 2 times, each 2h, and filter, merging filtrate, is evaporated to 40 DEG C of relative densities 1.15, obtains polysaccharide concentrated solution;
(2) in the polysaccharide concentrated solution of step (1), add volume fraction 98% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 60 times, acetone, anhydrous diethyl ether, 1 time is respectively washed successively to gained throw out, drying, obtains Crude polysaccharides;
(3) the Crude polysaccharides 0.5g getting step (2) is dissolved in 10mL water, is splined on HPD-600 type macroporous adsorptive resins, and with 250mL water elution, flow velocity is 1.0mL/min, collects elutriant, then is evaporated to 1/2 of elutriant original volume; Add volume fraction 98% ethanol and reach 80% to alcohol content, hold over night, with the dehydrated alcohol of weight of precipitate 60 times, acetone, anhydrous diethyl ether, 1 time is respectively washed successively to gained throw out, dry, obtain pilewort polysaccharide.
Pilewort polysaccharide of the present invention can be used for the preparation of medicine, and especially for strengthening immunity medicine, and obtain sufficient proof through test, related tests data is as follows:
Assay
Embodiment 1 gained pilewort polysaccharide is grey-brown powder, and polysaccharide yield is 15.63%, and measuring polysaccharide content through phend-sulphuric acid is 51.48%.
Pilewort polysaccharide strengthens the experiment of immunologic function
Experiment material
1. laboratory animal: Kunming kind cleaning grade healthy mice, body weight 18 ~ 22g, male and female half and half, are provided by laboratory animal room of Lukang Medical Co., Ltd., Shandong, conformity certification number: SCXK Shandong 20130001.
2. experimental drug: pilewort polysaccharide, Switzerland's dye liquor, lentinan film (Kaifeng pharmacy Group Co., Ltd), Cyclophosphamide for injection (SHANXI POWERDONE PHARMACEUTICAL.,LTD), Sodium Citrate, calcium chloride, sodium-chlor, Seignette salt, Repone K, glucose, dipotassium hydrogen phosphate, potassium primary phosphate, phenol red.
Two, experimental technique
1. pilewort polysaccharide impact that hypoimmunity mice hemolysin and hemolysis plaque are formed
Get Kunming mouse 300, about body weight 20g, male and female half and half, random point 6 groups, often organize 50, be respectively: blank group, model group, positive controls, pilewort polysaccharide 50mg/kg, 100mg/kg, 200mg/kg dosage group.Except blank group, all the other respectively organize equal abdominal injection concentration be the cyclophosphamide solution of 8mg/mL for three days on end, set up endoxan immunosuppression mouse model.Except naive mice immunologic function all reduces in 7 ~ 10 days.The sheep red blood cell (SRBC) normal saline suspension respectively organizing the equal abdominal injection 0.2mL/ of mouse 5% on the 1st day after modeling carries out immunity.1-7 days after modeling terminates, positive controls (lentinan 100mg/kg), pilewort polysaccharide 50mg/kg, 100mg/kg, 200mg/kg dosage component is not administered once according to corresponding dosage every day; Equal-volume normal saline solution is gavaged once a day to blank group, model group mouse, administration every day 1 time, continue medication 7 days.After 7th day administration 2h, mouse is plucked eyeball and gets blood, centrifugal, obtains upper serum, serum normal saline dilution 100 times.Serum dilution, the 0.5mL volume fraction 5% sheep red blood cell (SRBC) suspension of inhaling 1mL mix with 0.5mL volume fraction 10% guinea pig serum, after being placed in 37 DEG C of water-bath 30min, are placed in frozen water termination reaction, with the centrifugal 10min of 2500r/min.Separately establish not increase serum blank tube to compare, get supernatant liquor and survey absorbancy in 540nm place, obtain the hemolysin formational situation of each group of mouse, the results are shown in Table 1.
By pluck eyeball get the mouse after blood take off cervical vertebra put to death, solution takes spleen, homogenate, and adjusts splenocyte number, makes spleens cell number in splenocyte suspension be 5 × 10
6individual/mL.Inhale the splenocyte suspension 1mL after adjustment, then add 0.5mL volume fraction 0.2% sheep red blood cell (SRBC) suspension and volume fraction 10% guinea pig serum 0.5mL, mixing, puts 37 DEG C of water-bath 1h, with the centrifugal 10min of 2000r/min.Separately establish and do not add splenocyte suspension blank tube and compare, get supernatant liquor and survey absorbancy in 413nm place, obtain the hemolysis plaque cell formational situation of each group of mouse, the results are shown in Table 2.
Impact that table 1 pilewort polysaccharide is formed immunosuppressed mice hemolysin (
n=50)
Compare with model group, * represents P<0.05; * represents P<0.01
Impact that table 2 pilewort polysaccharide is formed immunosuppressed mice hemolysis plaque (
n=50)
Compare with model group, * represents P<0.05; * represents P<0.01
As can be seen from table 1-2, compared with blank group, the mouse hemolysin of model group and hemolysis plaque are formed and all significantly reduce, and illustrate that endoxan causes mouse immune inhibition modeling success.Compared with model group, pilewort polysaccharide 50mg/kg, 100mg/kg, 200mg/kg dosage group and positive controls all extremely significantly can suppress the formation (P<0.01) of mouse hemolysin and hemolysis plaque by Promote immunity.
2. pilewort polysaccharide is on the impact of hypoimmunity mice Peritoneal Macrophage Phagocytosis
The grouping of mouse, specification, modeling method, medication and dosage are all the same.In the 7th day sheep red blood cell (SRBC) physiological saline suspension solution 0.5mL/ to each group of equal abdominal injection volume fraction 5% of mouse only, administration after injection 2h, after gastric infusion 2h (after namely giving sheep red blood cell (SRBC) suspension 4h), mouse is taken off cervical vertebra to put to death, then abdominal injection Hank ' s application liquid 2.5mL, after gently rubbing mouse web portion, an aperture is cut off at mouse web portion, draw peritoneal fluid 2mL with liquid-transfering gun and be placed in plastic test tube, mixing, draw 20 μ L abdominal cavity drops in slide glass one end, push jack, and slide glass is placed in the porcelain dish being covered with wet gauze, 30min is incubated under 37 DEG C of conditions, attachment adhesive cell is washed away with physiological saline, drip the dyeing of blood cell staining liquid, water rinses, dry, that examines under a microscope Turnover of Mouse Peritoneal Macrophages engulfs situation, and calculate phagocytic percentage and phagocytic index, calculation formula is as follows, the results are shown in Table 3.
Phagocytic percentage (%)=(engulfing the scavenger cell number/scavenger cell sum of chicken red blood cell) × 100%
Sum/scavenger cell the sum of phagocytic index=macrophage phagocytic chicken red blood cell
Table 3 pilewort polysaccharide on the impact of hypoimmunity mice phagocytic function (
n=50)
Compare with model group, * represents P<0.05; * represents P<0.01
As can be seen from Table 3, compared with blank group, the mouse phagocytic percentage of model group and phagocytic index all significantly reduce, and illustrate that endoxan causes mouse immune inhibition modeling success.Compared with model group, pilewort polysaccharide 50m g/kg dosage group and positive controls all can form pole significance impact (P<0.01) to phagocytic percentage; 200m g/kg dosage group can form significance impact (P<0.05) to phagocytic percentage; 100mg/kg dosage group affects without significance phagocytic percentage.50mg/kg, 100mg/kg, 200mg/kg pilewort polysaccharide group and positive controls all can form pole significance impact (P<0.01) to phagocytic index.
Verify through experimentation on animals, the pilewort polyose composition extracted in the present invention has the activity of good enhancing immunity, possess the potentiality and actual promotional value that are developed as medicine or protective foods completely, this constituents can be used for the symptom improving the hypoimmunity caused by many reasons such as hypotrophy, unordered, overtired, the tumour patient chemicotherapies of living.Development research pilewort polysaccharide not only can widen the new prospect of Chinese medicine pilewort application, follows the creative contribution of era step to Chinese materia medica especially closely, has huge economy and social benefit.
Claims (7)
1. the pilewort polysaccharide extracted from pilewort, is characterized in that, the extracting method of described pilewort polysaccharide comprises the following steps:
(1) get pilewort medicinal powder, add the water of 10 ~ 40 times of pilewort medicinal powder weight, 60 ~ 100 DEG C are extracted 2 ~ 3 times, each 1 ~ 3h, and filter, merging filtrate, is evaporated to 40 DEG C of relative densities 1.10 ~ 1.20, obtains polysaccharide concentrated solution;
(2) in the polysaccharide concentrated solution of step (1), add volume fraction 95% ~ 100% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 50 ~ 100 times, acetone, anhydrous diethyl ether, 1 ~ 2 time is respectively washed successively to gained throw out, drying, obtains Crude polysaccharides;
(3) the Crude polysaccharides 0.2 ~ 1.0g getting step (2) is dissolved in 10mL water, be splined on HPD-600 type macroporous adsorptive resins, and with 200 ~ 300mL water elution, flow velocity is 0.5 ~ 1.5mL/min, collect elutriant, then be evaporated to the 1/2-2/3 of elutriant original volume; Add volume fraction 95% ~ 100% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 50 ~ 100 times, acetone, anhydrous diethyl ether, 1 ~ 2 time is respectively washed successively to gained throw out, dry, obtain pilewort polysaccharide.
2. pilewort polysaccharide according to claim 1, is characterized in that, the pilewort medicinal powder in step (1) was the powder of No. 2 pharmacopeia sieves.
3. pilewort polysaccharide according to claim 1, is characterized in that, the extracting method of described pilewort polysaccharide comprises the following steps:
(1) get pilewort medicinal powder, add the water of 10 times of pilewort medicinal powder weight, 60 DEG C are extracted 2 times, each 1h, and filter, merging filtrate, is evaporated to 40 DEG C of relative densities 1.10, obtains polysaccharide concentrated solution;
(2) in the polysaccharide concentrated solution of step (1), add volume fraction 95% ethanol and reach 80% to alcohol content, hold over night, gets bottom settlings, respectively washs 1 time successively to gained throw out with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether, drying, obtains Crude polysaccharides;
(3) the Crude polysaccharides 0.2g getting step (2) is dissolved in 10mL water, is splined on HPD-600 type macroporous adsorptive resins, and with 200mL water elution, flow velocity is 0.5mL/min, collects elutriant, then is evaporated to 1/2 of elutriant original volume; Add volume fraction 95% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether, 1 time is respectively washed successively to gained throw out, dry, obtain pilewort polysaccharide.
4. pilewort polysaccharide according to claim 1, is characterized in that, the extracting method of described pilewort polysaccharide comprises the following steps:
(1) get pilewort medicinal powder, add the water of 40 times of pilewort medicinal powder weight, 100 DEG C are extracted 3 times, each 3h, and filter, merging filtrate, is evaporated to 40 DEG C of relative densities 1.20, obtains polysaccharide concentrated solution;
(2) in the polysaccharide concentrated solution of step (1), add volume fraction 100% ethanol and reach 80% to alcohol content, hold over night, gets bottom settlings, respectively washs 2 times successively to gained throw out with the dehydrated alcohol of weight of precipitate 100 times, acetone, anhydrous diethyl ether, drying, obtains Crude polysaccharides;
(3) the Crude polysaccharides 1.0g getting step (2) is dissolved in 10mL water, is splined on HPD-600 type macroporous adsorptive resins, and with 300mL water elution, flow velocity is 1.5mL/min, collects elutriant, then is evaporated to 2/3 of elutriant original volume; Add volume fraction 100% ethanol and reach 80% to alcohol content, hold over night, get bottom settlings, with the dehydrated alcohol of weight of precipitate 100 times, acetone, anhydrous diethyl ether, 2 times are respectively washed successively to gained throw out, dry, obtain pilewort polysaccharide.
5. pilewort polysaccharide according to claim 1, is characterized in that, the extracting method of described pilewort polysaccharide comprises the following steps:
(1) get pilewort medicinal powder, add the water of 25 times of pilewort medicinal powder weight, 80 DEG C are extracted 2 times, each 2h, and filter, merging filtrate, is evaporated to 40 DEG C of relative densities 1.15, obtains polysaccharide concentrated solution;
(2) in the polysaccharide concentrated solution of step (1), add volume fraction 98% ethanol and reach 80% to alcohol content, hold over night, gets bottom settlings, respectively washs 1 time successively to gained throw out with the dehydrated alcohol of weight of precipitate 60 times, acetone, anhydrous diethyl ether, drying, obtains Crude polysaccharides;
(3) the Crude polysaccharides 0.5g getting step (2) is dissolved in 10mL water, is splined on HPD-600 type macroporous adsorptive resins, and with 250mL water elution, flow velocity is 1.0mL/min, collects elutriant, then is evaporated to 1/2 of elutriant original volume; Add volume fraction 98% ethanol and reach 80% to alcohol content, hold over night, with the dehydrated alcohol of weight of precipitate 60 times, acetone, anhydrous diethyl ether, 1 time is respectively washed successively to gained throw out, dry, obtain pilewort polysaccharide.
6. the application of pilewort polysaccharide as claimed in claim 1 in preparation enhancing immunologic function medicine.
7. the application of pilewort polysaccharide as claimed in claim 1 in preparation enhancing immunologic function protective foods.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105148258A (en) * | 2015-10-20 | 2015-12-16 | 杭州华津药业股份有限公司 | Composition and application thereof, and preparation containing composition |
| CN115636883A (en) * | 2022-09-05 | 2023-01-24 | 溯玄(上海)生物技术有限公司 | Plant purification component, preparation method, skin repair activity and application of plant purification component in preparation of skin repair cosmetics |
-
2015
- 2015-06-24 CN CN201510355723.8A patent/CN104892789A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| GERENCER, M等: "In vitro and in vivo anti-retroviral activity of the substance purified from the aqueous extract of Chelidonium majus L.", 《ANTIVIRAL RESEARCH》 * |
| SONG, JY等: "Immunomodulatory activity of protein-bound polysaccharide extracted from Chelidonium majus", 《ARCHIVES OF PHARMACAL RESEARCH》 * |
| 张杰: "河南中医学院硕士学位论文:白屈菜多糖提取纯化工艺及免疫活性研究", 《中国优秀硕士学位论文全文数据库》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105148258A (en) * | 2015-10-20 | 2015-12-16 | 杭州华津药业股份有限公司 | Composition and application thereof, and preparation containing composition |
| CN115636883A (en) * | 2022-09-05 | 2023-01-24 | 溯玄(上海)生物技术有限公司 | Plant purification component, preparation method, skin repair activity and application of plant purification component in preparation of skin repair cosmetics |
| CN115636883B (en) * | 2022-09-05 | 2024-04-26 | 广州梵之容化妆品有限公司 | Plant purification component, preparation method, skin repair activity and application of plant purification component in preparation of skin repair cosmetics |
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