CN104069155A - Application of oxytropis glabra flavonoid extract taken as bacteriostatic agent - Google Patents
Application of oxytropis glabra flavonoid extract taken as bacteriostatic agent Download PDFInfo
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- CN104069155A CN104069155A CN201410302341.4A CN201410302341A CN104069155A CN 104069155 A CN104069155 A CN 104069155A CN 201410302341 A CN201410302341 A CN 201410302341A CN 104069155 A CN104069155 A CN 104069155A
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- glabrous crazyweed
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention discloses an application of an oxytropis glabra flavonoid extract taken as a bacteriostatic agent. The experiment shows that the extract of chemical flavonoid ingredients in oxytropis glabra and a pharmaceutic preparation of the extract can be used for effectively inhibiting common pathogenic microorganisms such as staphylococcus aureus, bacillus subtilis, escherichia coli, proteusbacillus vulgaris and the like.
Description
Technical field
The pharmaceutical use that the present invention relates to the flavone extract of glabrous crazyweed, belongs to field of medicaments.
Background technology
Food processing at present and storage are very easily subject to the harm of the various bacteria such as Escherichia coli, staphylococcus aureus, bacillus subtilis, Bacillus proteus and mycete, unfavorable to food quality and consumer health.In order to kill and to suppress pathogenic bacteria, grow, the existing compositions such as chemical preservatives such as adding sodium benzoate, potassium sorbate that adopt suppress harmful microorganism growth more, but these chemical preservatives have potential harm to human body.
Meanwhile, at field of medicaments, because antibiotic abuse has caused the sharp increase of Resistant strain, the drug resistance of antibacterial and drug resistance infect has become current problem demanding prompt solution, and its solution is except rational use of drug, and the screening of new antimicrobial agent is also very important.
Glabrous crazyweed (Oxytropisglabra DC) is pulse family (Leguminosae) Genus Oxytropis Dc (Oxytropis) plant, widely distributed in Xinjiang, is a kind of important mongolian medicine and Kazakhstan medicine raw material.Flavone compound is one of effective ingredient of glabrous crazyweed, research both at home and abroad thinks that flavone compound has the effects such as antiinflammatory, mutation, blood pressure lowering, heat-clearing and toxic substances removing, calmness, diuresis, but up to the present, there are not research report and the patent application of glabrous crazyweed flavones preparation bacteriostasis.
Summary of the invention
The discovery that studies for a long period of time that applicant carries out, the flavone extract of glabrous crazyweed has the effect of the various common bacterias of extensive inhibition, and its fastbacteria is few, medicine source abundant, and untoward reaction is few, applies easy.
Based on above-mentioned discovery, applicant has completed technical scheme of the present invention.
First, the invention discloses the flavone extract of glabrous crazyweed as the application of antibacterial.
Wherein, above-mentioned application for antibacterial contained current common pathogenic bacteria microorganism, include but not limited to staphylococcus aureus, bacillus subtilis, escherichia coli, Bacillus proteus etc., applicant confirms by experiment for multiple-microorganism antibacterial, the flavone extract effect of glabrous crazyweed is all better than traditional sodium benzoate, potassium sorbate etc., and has good safety.
Wherein, the flavone extract of glabrous crazyweed can be bought from the market, also can extract voluntarily according to the record of document, such as glabrous crazyweed total flavone extracting process and different growing stages general flavone content (Luo Yang etc., Jiangsu agricultural sciences, 2011.03), the extraction process of the flavone extract of glabrous crazyweed has all been studied in research of grassland, Erdos glabrous crazyweed flavones content and allelopathy (Luo Yang, forestry in Beijing science, 2011) etc.
Preferably, the preparation method of the flavone extract of glabrous crazyweed comprises the steps:
1) by glabrous crazyweed raw material in organic solvent by microwave extraction, filtration, obtain glabrous crazyweed total extract after concentrated;
2) glabrous crazyweed total extract is crossed polyamide chromatographic column 10%～100% ethanol elution gradient elution, obtains the flavone extract of glabrous crazyweed after the ethanol elution concentrate drying of collection 80%～100%.
In said method, organic solvent used is preferably ethanol, and microwave extraction parameter can be adjusted as required.
The preferred microwave extraction parameter of the present invention is microwave power 300W, methanol concentration 100%, extraction time 120s, material ratio 1: 20.The parameter of polyamide separation and purification is: loading concentration 2.0mg/mL, and upper column flow rate is 3BV/h, and desorption concentration of alcohol is 80%～100%, and flow velocity is 3BV/h.
Extracting method of the present invention is not limited in above-mentioned parameter, suitably adjusts microwave power, extraction time, material ratio, loading concentration, flow velocity etc. and all it will be appreciated by those skilled in the art that, this adjustment only affects productive rate.
Because the flavone extract of glabrous crazyweed has the desirable ability of killing with anti-bacteria, the invention also discloses a kind of pharmaceutical composition with fungistatic effect, comprise that the flavone extract of glabrous crazyweed is as bacteriostatic active ingredients, and any available adjuvant pharmaceutically.
Above-mentioned adjuvant, with the dosage form significant correlation of pharmaceutical composition, those skilled in the art can, according to concrete preparation needs, adjust type and the consumption of adjuvant.Concrete preparation type can be the dosage form of the Orally-administrables such as tablet, pill, capsule, granule, powder, liquor, in these dosage forms, by pharmaceutic adjuvant combinations such as the flavone extract of glabrous crazyweed and one or more lubricants, disintegrating agent, stabilizing agent, dissolution aids, sugar-coat or gastric solubleness or enteric coatings, these adjuvants are all the pharmaceutic adjuvants of this area standard, and it will not go into details herein.
For example, when above-mentioned preparation is the fluid composition of oral administration, example emulsion, solution, suspensoid, syrup etc. are that the pharmaceutic adjuvants such as compositions and solubilizing agent, wetting agent, suspending agent, sweeting agent, flavoring agent, aromatic, antiseptic are used in combination.
Preferably, in described pharmaceutical composition, described adjuvant is beta-schardinger dextrin-(excipient), the flavone extract 3～5% that its consumption (mass percent) is glabrous crazyweed, beta-schardinger dextrin-95～97%.
For the ease of using, pharmaceutical composition can be added to the water and be mixed with solution, in this case, the concentration of pharmaceutical composition is 0.005g/mL～0.020g/mL, pH is 5.0～6.0.
By above-mentioned improvement, technical scheme tool of the present invention has the following advantages:
1) has a broad antifungal spectrum, fastbacteria is few;
2) medicine source is abundant, can gather materials on the spot;
3) effect extensively, except resisting pathogenic microbes effect, also has the pharmacological actions such as immunomodulating;
4) untoward reaction is few, applies easy.
The specific embodiment
Embodiment 1: the extraction preparation of glabrous crazyweed flavone
Take glabrous crazyweed sample, according to the ratio of material ratio 1:20g/mL, add the methanol microwave extraction 120s of volume fraction 100%, microwave power 300W, temperature 45 C, has extracted rear filtration, and filtrate adopts Rotary Evaporators concentrated, thermostatic drying chamber is dry, obtains glabrous crazyweed total extract; Glabrous crazyweed total extract water is fully dissolved; To 95% ethanol that adds 1/3 volume in chromatographic column, then polyamide is transferred in chromatographic column with ethanol, add 100% alcoholic solution (V/V), volume is 3 times of resin volume, flow velocity is 1 times of resin volume/min, then with deionized water, be washed till without carrying out the gradient elution of glabrous crazyweed total extract aqueous solution after alcohol taste, loading concentration 2.0mg/mL wherein, upper column flow rate is 3BV/h, ethanol elution with 10%～100%, collects 80%～100% ethanol elution, concentrated through Rotary Evaporators, freeze drier is dry
Obtain brown color powder and be glabrous crazyweed flavone extract.
Embodiment 2: the pharmaceutical preparation that contains glabrous crazyweed flavone extract, and can be according to following proportional arrangement
1) according to following ratio preparation: glabrous crazyweed flavone extract content 3%, excipient beta-schardinger dextrin-content 97%;
2) according to following ratio preparation: glabrous crazyweed flavone extract content 4%, excipient beta-schardinger dextrin-content 96%;
3) according to following ratio preparation: glabrous crazyweed flavone extract content 5%, excipient beta-schardinger dextrin-content 95%.Embodiment 3: the solution of the pharmaceutical preparation that contains glabrous crazyweed flavone extract, and can be according to following proportional arrangement
1) get the pharmaceutical preparation of embodiment 3 configurations 1, be added to the water, being diluted to concentration is the glabrous crazyweed flavone antibacterial solution of 0.005g/mL, uses food stage acetic acid to regulate the pH to 5.0 of antibacterial liquid;
2) get the pharmaceutical preparation of embodiment 3 configurations 2, be added to the water, being diluted to concentration is the glabrous crazyweed flavone antibacterial solution of 0.010g/mL, uses food stage acetic acid to regulate the pH to 5.4 of antibacterial liquid;
3) get the pharmaceutical preparation of embodiment 3 configurations 1, be added to the water, being diluted to concentration is the glabrous crazyweed flavone antibacterial solution of 0.020g/mL, uses food stage acetic acid to regulate the pH to 6.0 of antibacterial liquid;
Embodiment 4: the bacteriostatic experiment of glabrous crazyweed flavone extract
Get respectively the glabrous crazyweed flavone antibacterial solution of embodiment 4 configurations 1,2,3, the filter paper that to infiltrate diameter be 6mm carries out bacteriostatic test on the flat board that is coated with harmful microorganism bacterium liquid, cultivates 24h at 37 ℃, the antibacterial circle diameter of mensuration to various harmful bacterias, statistics maximal and minmal value; With the positive contrast of sodium benzoate, potassium sorbate that contains same active component consumption, add up equally maximal and minmal value.
Table 1: configuration 1 is for trying thing to harmful microbe inhibition
Table 2: configuration 2 is for trying thing to harmful microbe inhibition
Table 3: configuration 3 is for trying thing to harmful microbe inhibition
From above-mentioned experiment, can find out, under equal conditions and dosage, glabrous crazyweed flavones preparation fungistatic effect is better than sodium benzoate and potassium sorbate.
Embodiment 5: the zoopery of glabrous crazyweed flavone extract
Get 240 of empty stomach mices, male and female half and half, are divided into 24 groups at random by mice, and 10 every group, wherein 12 groups are used for infection of staphylococcus aureus, and 12 groups for streptococcal infection; By 1MLD (minimum lethal dose) bacterium amount every Mus lumbar injection staphylococcus aureus or streptococcus 0.5mL, prepare systemic infection model.In 12 test group, glabrous crazyweed flavone, azithromycin, each 4 dosage groups of erythromycin gavage treatment are treated.Glabrous crazyweed flavone extract dosage is respectively 8.4,11.2,14.9 and 20.0mg/kg body weight; Azithromycin dosage is respectively 26.6,35.4,47.3 and 63.0mg/kg body weight, and erythromycin dosage is respectively 53.8,71.7,95.6 and 127.5mg/kg body weight, respectively at 0h after infecting and each administration of 6h 1 time, each 0.5mL.Then apply DAS ver1.0 software kit, by Bliss method, calculate respectively median effective dose (ED50) and 95% fiducial limit thereof of glabrous crazyweed flavone, azithromycin and erythromycin for treating staphylococcus aureus and streptococcal infection mice.
Table 4 is the antibacterial activity in vivo to infecting mouse for examination thing
Result shows: glabrous crazyweed flavone has obvious antibacterial activity in vivo, and its curative effect to staphylococcus aureus is better than azithromycin and erythromycin, and the curative effect of streptococcus pyogenes is better than to erythromycin.
Get 60 of mices, male and female half and half, are divided into 4 test group at random, the glabrous crazyweed flavone of every group of single gavage various dose (5000,2500,1250,625mg/kg body weight), gavage amount 0.5mL.After administration, at once observe mice reaction, and record the death condition of respectively organizing mice in 7d, measure mouse blood and learn index, Biochemical Indices In Serum and organ coefficient.
Table 5 mice serum Biochemical Indexes result (X ± S)
Table 6 mouse blood is learned index determining result (X ± S)
Table 7 glabrous crazyweed flavone is on the impact of mice organs coefficient (X ± S)
From above-mentioned experimental result, can find out that mice searches for food the glabrous crazyweed flavone of 5000mg/kg body weight and have not yet to see abnormal clinical symptoms, mouse blood is learned index, Biochemical Indices In Serum and organ coefficient all within normal range.Glabrous crazyweed flavone has good safety to mice.
Claims (6)
1. the flavone extract of glabrous crazyweed is as the application of antibacterial.
2. according to the application of claim 1, it is characterized in that the preparation method of the flavone extract of glabrous crazyweed comprises the steps: 1) glabrous crazyweed raw material in organic solvent by microwave extraction, filtration, obtain glabrous crazyweed total extract after concentrated; 2) glabrous crazyweed total extract is crossed polyamide chromatographic column 10%～100% ethanol elution gradient elution, obtains the flavone extract of glabrous crazyweed after the ethanol elution concentrate drying of collection 80%～100%.
3. according to the application of claim 1, it is characterized in that the pathogenic bacteria microorganism suppressing comprises staphylococcus aureus, bacillus subtilis, escherichia coli, Bacillus proteus.
4. a pharmaceutical composition with fungistatic effect, is characterized in that the flavone extract that comprises glabrous crazyweed is as bacteriostatic active ingredients, and any available adjuvant pharmaceutically.
5. according to the pharmaceutical composition of claim 4, it is characterized in that described adjuvant is beta-schardinger dextrin-, the flavone extract 3～5% that its consumption is glabrous crazyweed, beta-schardinger dextrin-95～97%.
6. according to the pharmaceutical composition of claim 4, it is characterized in that dosage form is solution, concentration is 0.005g/mL～0.020g/mL, and pH is 5.0～6.0.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106727881A (en) * | 2017-03-15 | 2017-05-31 | 中国科学院西北高原生物研究所 | A kind of extracting method with anti-inflammatory activity general flavone from falcate crazyweed herb |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103202872A (en) * | 2013-04-08 | 2013-07-17 | 塔里木大学 | Preparation method and applications of flavonoid from Oxytropis glabra DC |
| CN103301181A (en) * | 2013-06-01 | 2013-09-18 | 塔里木大学 | Preparation method and application of oxytropis glabra daidzrin |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103202872A (en) * | 2013-04-08 | 2013-07-17 | 塔里木大学 | Preparation method and applications of flavonoid from Oxytropis glabra DC |
| CN103301181A (en) * | 2013-06-01 | 2013-09-18 | 塔里木大学 | Preparation method and application of oxytropis glabra daidzrin |
Non-Patent Citations (2)
| Title |
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| 于荣敏等: "小花棘豆化学成分的研究", 《JOURNAL OF INTEGRATIVE PLANT BIOLOGY 植物学报(英文版) 》 * |
| 惠永正编著: "《中药天然产物大全（4中天然产物）》", 31 January 2011, 上海科学技术出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106727881A (en) * | 2017-03-15 | 2017-05-31 | 中国科学院西北高原生物研究所 | A kind of extracting method with anti-inflammatory activity general flavone from falcate crazyweed herb |
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