CN104926958A - Method for extracting herba lycopi polysaccharide from herba lycopi and application of herba lycopi polysaccharide - Google Patents
Method for extracting herba lycopi polysaccharide from herba lycopi and application of herba lycopi polysaccharide Download PDFInfo
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Abstract
The invention relates to a method for extracting herba lycopi polysaccharide from herba lycopi and the application of the herba lycopi polysaccharide. The extraction and separation aims of herba lycopi polysaccharide components can be effectively achieved. According to the technical scheme, herba lycopi medicine decoction pieces are taken, water is added, extraction is carried out one to three times at the temperature of 70 DEG C to 90 DEG C, extraction is carried out for 1-3 h every time, filtering is carried out, filter liquor is condensed, ethyl alcohol of more than 95 percent is added into concentrated liquor till the alcohol content can be 75 percent to 85 percent, the concentrated liquor stands still overnight, filtering is carried out to obtain crude polysaccharide precipitate, and then HPD-100 macroreticular resin is used for purification. The herba lycopi polysaccharide can be used for development of drugs and healthcare products with antioxidant functions, the new application prospect of herba lycopi medicinal materials is developed, and the innovation on extraction and separation of the herba lycopi polysaccharide components is achieved.
Description
Technical field
The present invention relates to field of medicaments, particularly a kind of method and application thereof of extracting Herba Lycopi's polysaccharide from Herba Lycopi.
Background technology
Glucide is one of three major nutrient, and along with the develop rapidly of analytical technology and experimental technique, existing large quantity research show the polysaccharide separated from natural product have antitumor, anti-oxidant, reducing blood-fat, hypoglycemic, improve the pharmacologically actives such as immunizing power, the medicine of multiple polyose is applied clinically.
Herba Lycopi is the dry aerial parts of labiate hair leaf lycopus lucidus Lycopus lucidus Turcz var.hirtus Regel.Begin to be loaded in Shennong's Herbal, history tree is all on the books.Herba Lycopi's bitter, pungent, tepor, returns liver, the spleen channel.There is merit that is promoting blood circulation and removing blood stasis, line water detumescence.For menoxenia, through closing, dysmenorrhoea, postpartum blood stasis is suffered from abdominal pain, the diseases such as oedema, and Herba Lycopi mainly containing triterpene acids, phenolic acids, flavonoid, volatile oil, carbohydrate etc., and has no report to the extraction and isolation of Herba Lycopi's polysaccharide component and the research of pharmacologically active.
Summary of the invention
For above-mentioned situation, for solving the defect of prior art, the object of the present invention is just to provide a kind of method and application thereof of from Herba Lycopi, extracting Herba Lycopi's polysaccharide, effectively can solve the extraction and isolation problem of Herba Lycopi's polysaccharide component.
The technical scheme that the present invention solves is, comprise the following steps: 1) get Herba Lycopi's pharmaceutical decocting piece, add the water of Herba Lycopi's pharmaceutical decocting piece weight 20 ~ 30 times, 70 ~ 90 DEG C are extracted 1 ~ 3 time, each 1 ~ 3h, filter, when filtrate is concentrated into 60 DEG C, relative density is the polysaccharide concentrated solution of 1.10 ~ 1.25, adds the ethanol that volumetric concentration is more than 95%, makes alcohol content reach volumetric concentration 75% ~ 85%, hold over night, filter, obtain throw out, throw out absolute ethanol washing is colourless to washings, drying, obtains Herba Lycopi's Crude polysaccharides;
2) getting Herba Lycopi's Crude polysaccharides 0.3 ~ 0.5g is dissolved in 10ml water, be splined on HPD-100 macroporous adsorptive resins, with 200 ~ 400mL water elution, flow velocity is 0.5 ~ 1.5ml/min, merge effluent liquid, during reconcentration to 40 DEG C, relative density is the medicinal extract of 1.02 ~ 1.06, add the ethanol that volumetric concentration is more than 95%, reach volumetric concentration 80% to alcohol content, hold over night, filter, obtain throw out, throw out respectively washs 1 ~ 2 time with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether successively, dry, obtains refined polysaccharide.
Herba Lycopi's polysaccharide of the present invention can be used for the medicine of anti-oxidant function and the exploitation of healthcare products, opens up the application prospect that Herba Lycopi's medicinal material is new, be Herba Lycopi's polysaccharide component extraction and isolation on innovation.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described in further detail.
Embodiment 1
1) get Herba Lycopi's pharmaceutical decocting piece, add the water of Herba Lycopi's pharmaceutical decocting piece weight 30 times, 70 DEG C are extracted 2 times, each 2h, filters, and when filtrate is concentrated into 60 DEG C, relative density is the polysaccharide concentrated solution of 1.12, add the ethanol that volumetric concentration is 95%, be volumetric concentration 75% to solution alcohol content, hold over night, filters, obtain throw out, throw out absolute ethanol washing is colourless to washings, dry, obtains Herba Lycopi's Crude polysaccharides;
2) get Herba Lycopi's Crude polysaccharides 0.3g to be dissolved in 10ml water, be splined on HPD-100 macroporous adsorptive resins, use 300mL water elution, flow velocity is 1.5ml/min, merges effluent liquid, the medicinal extract of relative density 1.03 during reconcentration to 40 DEG C, add the ethanol that volumetric concentration is 95%, reach volumetric concentration 80% to alcohol content, hold over night, filter, obtain throw out, throw out respectively washs 1 time with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether successively, dry, obtains refined polysaccharide.
Embodiment 2
1) get Herba Lycopi's pharmaceutical decocting piece, add the water of Herba Lycopi's pharmaceutical decocting piece weight 25 times, 80 DEG C are extracted 3 times, each 1h, filters, and when filtrate is concentrated into 60 DEG C, relative density is the polysaccharide concentrated solution of 1.20, add the ethanol that volumetric concentration is 97%, make alcohol content reach volumetric concentration 80%, hold over night, filter, obtain throw out, throw out absolute ethanol washing is colourless to washings, dry, obtains Herba Lycopi's Crude polysaccharides;
2) get Herba Lycopi's Crude polysaccharides 0.4g to be dissolved in 10ml water, be splined on HPD-100 macroporous adsorptive resins, use 250mL water elution, flow velocity is 1.0ml/min, and merge effluent liquid, during reconcentration to 40 DEG C, relative density is the medicinal extract of 1.04, add the ethanol that volumetric concentration is 97%, reach volumetric concentration 80% to alcohol content, hold over night, filter, obtain throw out, throw out respectively washs 2 times with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether successively, dry, obtains refined polysaccharide.
Embodiment 3
1) get Herba Lycopi's pharmaceutical decocting piece, add the water of Herba Lycopi's pharmaceutical decocting piece weight 20 times, 90 DEG C are extracted 3 times, each 3h, filters, and when filtrate is concentrated into 60 DEG C, relative density is the polysaccharide concentrated solution of 1.25, add the ethanol that volumetric concentration is 100%, make alcohol content reach volumetric concentration 85%, hold over night, filter, obtain throw out, throw out absolute ethanol washing is colourless to washings, dry, obtains Herba Lycopi's Crude polysaccharides;
2) get Herba Lycopi's Crude polysaccharides 0.5g to be dissolved in 10ml water, be splined on HPD-100 macroporous adsorptive resins, use 200mL water elution, flow velocity is 0.5ml/min, and merge effluent liquid, during reconcentration to 40 DEG C, relative density is the medicinal extract of 1.05, add the ethanol that volumetric concentration is 95%, reach volumetric concentration 80% to alcohol content, hold over night, filter, obtain throw out, throw out respectively washs 2 times with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether successively, dry, obtains refined polysaccharide.
Described Herba Lycopi's medicine materical crude slice is stem and the leaf of Herba Lycopi 1 ~ 2cm length.
Described water is one or more of tap water, pure water, distilled water or deionized water.
Herba Lycopi's polysaccharide that the inventive method is extracted has oxidation resistant effect, animal experiment sufficient proof, and concrete test situation is as follows:
1. test materials
1.1 experimental animals: Kunming kind no-special pathogen level (SPF) healthy mice, male and female half and half, in 4 ~ 6 week age, body weight 18 ~ 22g, purchased from Henan Province's animal experimental center.
1.2 experiment reagents: Herba Lycopi's polysaccharide of the present invention; Vc effervescent tablet (federalism pharmaceutical factory company limited); Total protein quantification kit, total number born (T-SOD) test kit, catalase (CAT) test kit, Glutathione peroxidase (GSH-PX) test kit, mda (MDA) test kit all build up Bioengineering Research Institute purchased from Nanjing.
2. test method
Mouse is after animal feeding room adapts to one week, and 72 mouse are divided into 6 groups, often organizes male and female half and half.First group, gavage physiological saline 0.1mL/ (10gd), is blank group.Second group, abdominal injection D-semi-lactosi 150mg/ (kgd), gavage physiological saline 0.1mL/ (10gd) is model group.3rd group, abdominal injection D-semi-lactosi 150mg/ (kgd), and gavage vitamins C 150mg/ (kgd) are Vc group.4th group, abdominal injection D-semi-lactosi 150mg/ (kgd), and gavage Herba Lycopi polysaccharide 100mg/ (kgd) are low dose group.5th group, abdominal injection D-semi-lactosi 150mg/ (kgd), and gavage Herba Lycopi polysaccharide 200mg/ (kgd) are middle dosage group.6th group, abdominal injection D-semi-lactosi 150mg/ (kgd), gavage Herba Lycopi polysaccharide 300mg/ (kgd) is high dose group.Vc and Herba Lycopi's polysaccharide are the aqueous solution, D-semi-lactosi physiological saline solution, and mouse stomach volume is 0.1mL/ (10gd), after three weeks, fasting 12 hours, mouse takes off neck and puts to death, quick taking-up liver, brain, its quality of accurate title, adds the manual homogenate in frozen water of physiological saline glass homogenizer by quality and volume ratio 1 ﹕ 9 (g ﹕ mL), is the tissue homogenate of 10% evenly, 3000 revs/min, after centrifugal 10min, get supernatant liquor, freezing for subsequent use.Operate by the specification sheets of test kit, measure absorbancy with ultraviolet-visible pectrophotometer, calculate each desired value.
3. test-results
Table 1 Herba Lycopi Polysaccharides on Mice brain and liver tissues T-SOD vigor affect result table
Note: compare * * with model group be p<0.01, * is p<0.05
As can be seen from Table 1, the T-SOD vigor of model group Mice brain tissues and hepatic tissue is all remarkable in blank group, shows that the low drag success of total number born vigor made by abdominal injection D-semi-lactosi 150mg/ (kgd).
Also can find out, the T-SOD vigor of Herba Lycopi's polysaccharide and brain and hepatic tissue is positive dose relationship.The middle and high dosage group of Herba Lycopi's polysaccharide and Vc group all can promote liver, cerebral tissue T-SOD vigor (P<0.01) extremely significantly.
Table 2 Herba Lycopi Polysaccharides on Mice brain and liver tissues GSH-PX vigor affect result table
Note: compare * * with model group be p<0.01, * is p<0.05
As can be seen from Table 2, the GSH-PX vigor of model group Mice brain tissues and hepatic tissue is all remarkable in blank group, shows that the low drag success of Glutathione peroxidase (GSH-PX) vigor made by abdominal injection D-semi-lactosi 150mg/ (kgd).
Also can find out, Glutathione peroxidase (GSH-PX) vigor of Herba Lycopi's polysaccharide and brain and hepatic tissue is positive dose relationship.The basic, normal, high dosage group of Herba Lycopi's polysaccharide and Vc group all can promote murine liver tissue Glutathione peroxidase vigor (P<0.01) extremely significantly, Herba Lycopi's polysaccharide high dose group and Vc group can promote Mice brain tissues Glutathione peroxidase vigor (P<0.01) extremely significantly, and dosage group significantly can promote Mice brain tissues Glutathione peroxidase vigor (P<0.05) in Herba Lycopi's polysaccharide, low dose group compared with model group, without remarkable effect.
Table 3 Herba Lycopi Polysaccharides on Mice brain and liver tissues CAT vigor affect result table
Note: compare * * with model group be p<0.01, * is p<0.05
As can be seen from Table 3, the CAT vigor of model group Mice brain tissues and hepatic tissue is all remarkable in blank group, shows that the low drag success of catalase (CAT) vigor made by abdominal injection D-semi-lactosi 150mg/ (kgd).
Also can find out, catalase (CAT) vigor of Herba Lycopi's polysaccharide and brain and hepatic tissue is positive dose relationship.The basic, normal, high dosage group of Herba Lycopi's polysaccharide and Vc group all can promote Mice brain tissues catalase (CAT) vigor (P<0.01) extremely significantly, the middle and high dosage group of Herba Lycopi's polysaccharide and Vc group can promote murine liver tissue catalase (CAT) vigor (P<0.01) extremely significantly, and Herba Lycopi's polysaccharide low dose group significantly can promote murine liver tissue catalase (CAT) vigor (P<0.05)
Table 4 Herba Lycopi Polysaccharides on Mice brain and liver tissues MDA content affect result table
Note: compare * * with model group be p<0.01, * is p<0.05
As shown in Table 4, the MDA content of model group Mouse Liver, cerebral tissue is all significantly higher than blank group, shows abdominal injection D-semi-lactosi 150mg/ (kgd) modeling success.
Compare with model group, the middle and high dosage group of Herba Lycopi's polysaccharide and Vc group extremely significantly can reduce Mice brain tissues MDA content (P<0.01), Herba Lycopi's polysaccharide low dose group significantly can reduce Mice brain tissues MDA content (P<0.05), Herba Lycopi's polysaccharide is low, high dose group significantly can reduce murine liver tissue MDA content (P<0.05), but middle dosage group and the effect of Vc group not obvious.
Can draw from above four forms, model group has pole significant difference (p<0.01) with the blank Testing index organized, and namely abdominal injection D-semi-lactosi successfully causes aging model; Vc group compares with model group and can obtain, and outside Vc does not improve significantly to the content of hepatic tissue MDA, all has extremely significant improvement result (p<0.01) to other index of liver cerebral tissue; And Herba Lycopi's polysaccharide high dose group is compared with model group, belong to except significant difference (p<0.05) except to the content reducing effect of hepatic tissue MDA, to the improvement result of other indices, all there is pole significant difference (p<0.01).
The aging of human body, extremely, the strong oxidizing property of free radical, can damage tissue and the cell of body in the oxidation institute mainly caused due to free radical, and then cause aging effect and multiple chronic disease, as senile dementia, cardiovascular and cerebrovascular disease, cataract, Parkinson's disease, diabetes, tumour etc.Therefore, the resistance of oxidation wanting good health and a long life enhancing body is antidotal important step.
Results of animal shows; the Herba Lycopi's polysaccharide extracted in the present invention can improve the vigor of the antioxidases such as body SOD, GSH-Px, CAT; thus promote the elimination of machine interior free yl; alleviate lipid peroxidation to occur; protection microbial film; thus be expected to be developed to anti-oxidant and anti-ageing class healthcare products or medicine, open up the application prospect that Herba Lycopi's medicinal material is new, have actual promotional value.
Claims (7)
1. from Herba Lycopi, extract a method for Herba Lycopi's polysaccharide, it is characterized in that, comprise the following steps:
1) get Herba Lycopi's pharmaceutical decocting piece, add the water of Herba Lycopi's pharmaceutical decocting piece weight 20 ~ 30 times, 70 ~ 90 DEG C are extracted 1 ~ 3 time, each 1 ~ 3h, filters, and when filtrate is concentrated into 60 DEG C, relative density is the polysaccharide concentrated solution of 1.10 ~ 1.25, add the ethanol that volumetric concentration is more than 95%, make alcohol content reach volumetric concentration 75% ~ 85%, hold over night, filter, obtain throw out, throw out absolute ethanol washing is colourless to washings, dry, obtains Herba Lycopi's Crude polysaccharides;
2) getting Herba Lycopi's Crude polysaccharides 0.3 ~ 0.5g is dissolved in 10ml water, be splined on HPD-100 macroporous adsorptive resins, with 200 ~ 400mL water elution, flow velocity is 0.5 ~ 1.5ml/min, merge effluent liquid, during reconcentration to 40 DEG C, relative density is the medicinal extract of 1.02 ~ 1.06, add the ethanol that volumetric concentration is more than 95%, reach volumetric concentration 80% to alcohol content, hold over night, filter, obtain throw out, throw out respectively washs 1 ~ 2 time with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether successively, dry, obtains refined polysaccharide.
2. the method extracting Herba Lycopi's polysaccharide from Herba Lycopi according to claim 1, is characterized in that, comprise the following steps:
1) get Herba Lycopi's pharmaceutical decocting piece, add the water of Herba Lycopi's pharmaceutical decocting piece weight 30 times, 70 DEG C are extracted 2 times, each 2h, filters, and when filtrate is concentrated into 60 DEG C, relative density is the polysaccharide concentrated solution of 1.12, add the ethanol that volumetric concentration is 95%, be volumetric concentration 75% to solution alcohol content, hold over night, filters, obtain throw out, throw out absolute ethanol washing is colourless to washings, dry, obtains Herba Lycopi's Crude polysaccharides;
2) get Herba Lycopi's Crude polysaccharides 0.3g to be dissolved in 10ml water, be splined on HPD-100 macroporous adsorptive resins, use 300mL water elution, flow velocity is 1.5ml/min, merges effluent liquid, the medicinal extract of relative density 1.03 during reconcentration to 40 DEG C, add the ethanol that volumetric concentration is 95%, reach volumetric concentration 80% to alcohol content, hold over night, filter, obtain throw out, throw out respectively washs 1 time with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether successively, dry, obtains refined polysaccharide.
3. the method extracting Herba Lycopi's polysaccharide from Herba Lycopi according to claim 1, is characterized in that, comprise the following steps:
1) get Herba Lycopi's pharmaceutical decocting piece, add the water of Herba Lycopi's pharmaceutical decocting piece weight 25 times, 80 DEG C are extracted 3 times, each 1h, filters, and when filtrate is concentrated into 60 DEG C, relative density is the polysaccharide concentrated solution of 1.20, add the ethanol that volumetric concentration is 97%, make alcohol content reach volumetric concentration 80%, hold over night, filter, obtain throw out, throw out absolute ethanol washing is colourless to washings, dry, obtains Herba Lycopi's Crude polysaccharides;
2) get Herba Lycopi's Crude polysaccharides 0.4g to be dissolved in 10ml water, be splined on HPD-100 macroporous adsorptive resins, use 250mL water elution, flow velocity is 1.0ml/min, and merge effluent liquid, during reconcentration to 40 DEG C, relative density is the medicinal extract of 1.04, add the ethanol that volumetric concentration is 97%, reach volumetric concentration 80% to alcohol content, hold over night, filter, obtain throw out, throw out respectively washs 2 times with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether successively, dry, obtains refined polysaccharide.
4. the method extracting Herba Lycopi's polysaccharide from Herba Lycopi according to claim 1, is characterized in that, comprise the following steps:
1) get Herba Lycopi's pharmaceutical decocting piece, add the water of Herba Lycopi's pharmaceutical decocting piece weight 20 times, 90 DEG C are extracted 3 times, each 3h, filters, and when filtrate is concentrated into 60 DEG C, relative density is the polysaccharide concentrated solution of 1.25, add the ethanol that volumetric concentration is 100%, make alcohol content reach volumetric concentration 85%, hold over night, filter, obtain throw out, throw out absolute ethanol washing is colourless to washings, dry, obtains Herba Lycopi's Crude polysaccharides;
2) get Herba Lycopi's Crude polysaccharides 0.5g to be dissolved in 10ml water, be splined on HPD-100 macroporous adsorptive resins, use 200mL water elution, flow velocity is 0.5ml/min, and merge effluent liquid, during reconcentration to 40 DEG C, relative density is the medicinal extract of 1.05, add the ethanol that volumetric concentration is 95%, reach volumetric concentration 80% to alcohol content, hold over night, filter, obtain throw out, throw out respectively washs 2 times with the dehydrated alcohol of weight of precipitate 50 times, acetone, anhydrous diethyl ether successively, dry, obtains refined polysaccharide.
5. the method extracting Herba Lycopi's polysaccharide from Herba Lycopi according to claim 1 or 2-4, is characterized in that, described Herba Lycopi's medicine materical crude slice is stem and the leaf of Herba Lycopi 1 ~ 2cm length; Described water is one or more of tap water, pure water, distilled water or deionized water.
6. Herba Lycopi's polysaccharide that prepared by claim 1 or the method described in 2-4 is preparing the application in anti-oxidation medicine or antioxidant health-care product.
7. Herba Lycopi's polysaccharide that prepared by method according to claim 5 is preparing the application in anti-oxidation medicine or antioxidant health-care product.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108029727A (en) * | 2018-01-15 | 2018-05-15 | 广州聚澜健康产业研究院有限公司 | One kind smears tea multi-layer steamed bread and preparation method thereof |
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| CN118319939A (en) * | 2024-06-17 | 2024-07-12 | 长春中医药大学 | Application of herba lycopi polysaccharide in treatment of premature ovarian failure |
| CN118319939B (en) * | 2024-06-17 | 2024-10-15 | 长春中医药大学 | Application of herba lycopi polysaccharide in treatment of premature ovarian failure |
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