CN101019931A - Natural antibiotic composition extracted from crinite seed and its extraction process - Google Patents
Natural antibiotic composition extracted from crinite seed and its extraction process Download PDFInfo
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- CN101019931A CN101019931A CNA2007100776756A CN200710077675A CN101019931A CN 101019931 A CN101019931 A CN 101019931A CN A2007100776756 A CNA2007100776756 A CN A2007100776756A CN 200710077675 A CN200710077675 A CN 200710077675A CN 101019931 A CN101019931 A CN 101019931A
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- semen caesalpiniae
- caesalpiniae minacis
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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 present invention discloses one kind of natural antibiotic composition extracted from crinite seed and its extraction process. The extraction process includes crushing dry crinite seed and 40-200 mesh sieving, soaking in solvent for 1-5 times of 12-24 hr each, vibrating in shaking table of rotation speed 180-250 rpm at 30-40 deg.c for 12-24 hr, suction filtering to obtain extracted liquid, and drying to recover solvent and obtain the composition. The solvent is water, 95 % concentration alcohol solution or ethyl acetate. The natural antibiotic composition has excellent bactriostasis on staphylococcus aureus, colibacillus, fecal streptococcus, etc.
Description
Technical field
The present invention relates to bactericidal composition, relate in particular to a kind of natural antibacterial composition that from Semen Caesalpiniae Minacis, extracts, also relate to its extracting method simultaneously.
Background technology
Antibacterials are the widest class medicines of clinical practice, but the application of this class medicine exists many problems at present, the rapid rising of dysbacteriosis, superinfection and nosocomial infection and allergy etc., the especially bacterial drug resistance that causes as drug resistance, toxicity, antibacterials.Making does not have effective medicine to cause very big threat to human health for selecting for use to the severe infections due to this class bacterium.Therefore, people are developed the emphasis of new antibacterials as antibacterials research from natural resources at present.
Semen Caesalpiniae Minacis is called FRUCTUS NELUMBINIS, Semen Nelumbinis, shell Semen Nelumbinis, belt leather Semen Nelumbinis again.Derive from the mature fruit of nymphaeaceae plant lotus Nelumbo nucifera Gaertn.English name: NELUMBINIS LB this product is oval, and two ends are point slightly, about long 1.8cm, and the about 1.2cm of diameter.The shell grey black, surface smoothing, an end has little round concave point.The other end has small short handle, and the thick about 1mm of shell includes 1 piece in Semen Nelumbinis.Matter is hard, is difficult for breaking.Semen Caesalpiniae Minacis has the effect of the pyretic toxicity that strengthens the spleen and stomach, dispels, the relieving restlessness that clears away heart-fire, and can be used for alleviation and treats symptoms such as cold and hot vomiting, food are not following, stranguria with turbid discharge.
Summary of the invention
Purpose of the present invention overcome above-mentioned shortcoming and provide a kind of staphylococcus aureus (staphylococcus aureus), escherichia coli (Escherichia coli), streptococcus faecalis (Staphylococcus faecalis), Aspergillus flavus (Aspergillus flavus), bacillus pyocyaneus (Pseudomonas aeruginosa), micrococcus luteus (Micrococcus luteus) are had the inhibiting natural antibacterial composition that extracts from Semen Caesalpiniae Minacis.
Another object of the present invention provides the extracting method of the natural antibacterial composition that extracts from Semen Caesalpiniae Minacis.
The natural antibacterial composition that extracts from Semen Caesalpiniae Minacis of the present invention is characterized in that:
Exsiccant Semen Caesalpiniae Minacis was pulverized the 40-200 mesh sieve, added solvent in the ratio of 1-5g/10ml, soaks 1-5 time after, 12-24h at every turn, placing temperature is that 30-40 ℃, rotating speed are to shake 12-24h on the constant temperature shaking table of 180-250r/min, sucking filtration obtains extracting solution; Reclaim the solvent after drying, promptly.
Wherein: solvent is water, ethanol or ethyl acetate.
The extracting method of the natural antibacterial composition that from Semen Caesalpiniae Minacis, extracts, it is characterized in that: exsiccant Semen Caesalpiniae Minacis was pulverized the 40-200 mesh sieve, ratio in 1-5g/10ml adds solvent, after soaking 1-5 time, each 12-24h, placing temperature is that 30-40 ℃, rotating speed are to shake 12-24h on the constant temperature shaking table of 180-250r/min, and sucking filtration obtains extracting solution; Reclaim the solvent after drying, promptly.
The extracting method of the above-mentioned natural antibacterial composition that from Semen Caesalpiniae Minacis, extracts, the method that wherein reclaims the solvent after drying is: when solvent is water ,-80 ℃ of freezing 12-24h vacuum freeze drier dryings; When solvent was ethanol, 40 ℃ of decompression rotations of alcoholic extract concentrate removed ethanol ,-80 ℃ of freezing 12-24h vacuum freeze drier dryings; When solvent was ethyl acetate, 45 ℃ of concentrating under reduced pressure of acetic acid ethyl acetate extract were removed ethyl acetate.
The present invention compared with prior art passes through and experiment showed, that the aqueous solvent extract has effect preferably to micrococcus luteus, and minimum inhibitory concentration is 6.25mg/ml; To staphylococcus aureus, the minimum inhibitory concentration of Aspergillus flavus is 12.5mg/ml; To escherichia coli, the minimum inhibitory concentration of bacillus pyocyaneus is 25mg/ml; Minimum inhibitory concentration to enterococcus faecalis is 50mg/ml.The alcohol solvent extract is to staphylococcus aureus, escherichia coli, and micrococcus luteus, the inhibitory action of bacillus pyocyaneus is better, and minimum inhibitory concentration is 12.5mg/ml; The inhibition effect for the examination bacterium to other is poor slightly, is 25mg/ml to the minimum inhibitory concentration of enterococcus faecalis and Aspergillus flavus.The ethyl acetate solvent extract has effect preferably to Aspergillus flavus, and minimum inhibitory concentration is 6.25mg/ml; To other for the examination bacterium the inhibition effect slightly a little less than, to staphylococcus aureus, the minimum inhibitory concentration of bacillus pyocyaneus is 25mg/ml; To escherichia coli, the minimum inhibitory concentration of micrococcus luteus and enterococcus faecalis is 50mg/ml.Illustrate that the different solvents extract of Semen Caesalpiniae Minacis all has the activity of bacteria growing inhibiting.For the further development and use of Semen Caesalpiniae Minacis with seek new natural antibacterial medicine and provide the foundation.
The specific embodiment
Below further prove beneficial effect of the present invention by bacteriostatic activity experiment:
Embodiment 1
Exsiccant Semen Caesalpiniae Minacis was pulverized 40 mesh sieves, take by weighing 150g, the water that adds 1000ml, after soaking 2 times, each 12h, placing temperature is that 37 ℃, rotating speed are to shake 12h on the constant temperature shaking table of 200r/min, sucking filtration obtains extracting solution, with the dry 24h of vacuum freeze drier, obtain dry product behind-80 ℃ of freezing 12h.
Get the dry product 0.3g of Semen Caesalpiniae Minacis water extract, be dissolved in respectively among the 3mL DMF, cross biofilter, become 1: 2 with LB fluid medium doubling dilution respectively as stock solution, 1: 4,1: 8,1: 16,1: 32,1: 64,1: 128,1: 256 each 8 concentration, promptly contain original liquid concentration and be respectively 50,25,2.5,6.25,3.1,1.5,0.75,0.375mg/mL, every pipe 300ul.Each concentration is established three parallel pipes, and to establish the LB fluid medium that does not contain extract respectively be control tube.In each pipe, add good bacteria suspension (OD of 600nm place value the is about 1.0) 30uL of prepared beforehand then, be put in behind the mixing in 37 ℃ the incubator and cultivate 24h.Get 0.1ml liquid coating nutrient agar panel from each test tube respectively, three flat boards of every gradient coating, whole process repeats twice, has determined whether bacterial growth behind the 24h.The least concentration that suppresses bacteria growing to be measured with medicine is minimal inhibitory concentration (MIC).Experimental result is as follows:
| Extract | Strain | Concentration (mg/ml) | ||||||
| 50 | 25 | 12.5 | 6.25 | 3.13 | 1.57 | 0.79 | ||
| Water extract | Staphylococcus aureus | - | - | - | + + | + + | + + | + + |
| Escherichia coli | - | - | + | + + | + + | + + | + + | |
| Bacillus pyocyaneus | - | - | + | + | + + | + + | + + | |
| Micrococcus luteus | - | - | - | - | + + | + + | + + | |
| Enterococcus faecalis | - | + | + | + | + + | + + | + + | |
| Aspergillus flavus | - | - | - | + | + + | + + | + + |
Embodiment 2
Exsiccant Semen Caesalpiniae Minacis was pulverized 120 mesh sieves, take by weighing 150g, add 1000ml, 95% ethanol, soak 3 times after, each 18h, placing temperature is that 30 ℃, rotating speed are to shake 18h on the constant temperature shaking table of 250r/min, and sucking filtration obtains extracting solution, and 40 ℃ of decompression rotations concentrate and remove ethanol,-80 ℃ of freezing 12h obtain dry product with the dry 24h of vacuum freeze drier.
Get the extract dry product 0.3g of Semen Caesalpiniae Minacis 95% ethanol, be dissolved in respectively among the 3mL DMF, cross biofilter, become 1: 2 with LB fluid medium doubling dilution respectively as stock solution, 1: 4,1: 8,1: 16,1: 32,1: 64,1: 128,1: 256 each 8 concentration, promptly contain original liquid concentration and be respectively 50,25,12.5,6.25,3.1,1.5,0.75,0.375mg/mL, every pipe 300ul.Each concentration is established three parallel pipes, and to establish the LB fluid medium that does not contain extract respectively be control tube.In each pipe, add good bacteria suspension (OD of 600nm place value the is about 1.0) 30uL of prepared beforehand then, be put in behind the mixing in 37 ℃ the incubator and cultivate 24h.Get 0.1ml liquid coating nutrient agar panel from each test tube respectively, three flat boards of every gradient coating, whole process repeats twice, has determined whether bacterial growth behind the 24h.The least concentration that suppresses bacteria growing to be measured with medicine is minimal inhibitory concentration (MIC).Experimental result is as follows:
| Extract | Strain | Concentration (mg/ml) | ||||||
| 50 | 25 | 12.5 | 6.25 | 3.13 | 1.57 | 0.79 | ||
| Alcoholic extract | Staphylococcus aureus | - | - | - | + | + | ++ | ++ |
| Escherichia coli | - | - | - | ++ | ++ | ++ | ++ | |
| Bacillus pyocyaneus | - | - | - | + | ++ | ++ | ++ | |
| Micrococcus luteus | - | - | - | + | ++ | ++ | ++ | |
| Enterococcus faecalis | - | - | + | + | ++ | ++ | ++ | |
| Aspergillus flavus | - | - | + | ++ | ++ | ++ | ++ | |
Embodiment 3
Exsiccant Semen Caesalpiniae Minacis is pulverized 200 mesh sieves, took by weighing 150g, added the ethyl acetate of 1000ml, after soaking 5 times, each 24h, placing temperature is that 40 ℃, rotating speed are to shake 24h on the constant temperature shaking table of 180r/min, sucking filtration obtains extracting solution, and 45 ℃ of concentrating under reduced pressure are removed ethyl acetate, obtain dry product.
Get the dry product 0.3g of Semen Caesalpiniae Minacis ethyl acetate extract, be dissolved in respectively among the 3mL DMF, cross biofilter, become 1: 2 with LB fluid medium doubling dilution respectively as stock solution, 1: 4,1: 8,1: 16,1: 32,1: 64,1: 128,1: 256 each 8 concentration, promptly contain original liquid concentration and be respectively 50,25,2.5,6.25,3.1,1.5,0.75,0.375mg/mL, every pipe 300ul.Each concentration is established three parallel pipes, and to establish the LB fluid medium that does not contain extract respectively be control tube.In each pipe, add good bacteria suspension (OD of 600nm place value the is about 1.0) 30uL of prepared beforehand then, be put in behind the mixing in 37 ℃ the incubator and cultivate 24h.Get 0.1ml liquid coating nutrient agar panel from each test tube respectively, three flat boards of every gradient coating, whole process repeats twice, has determined whether bacterial growth behind the 24h.The least concentration that suppresses bacteria growing to be measured with medicine is minimal inhibitory concentration (MIC).Experimental result is as follows:
| Extract | Strain | Concentration (mg/ml) | ||||||
| 50 | 25 | 12.5 | 6.25 | 3.13 | 1.57 | 0.79 | ||
| Ethyl acetate is carried thing | Staphylococcus aureus | - | - | + | + + | + + | + + | + + |
| Escherichia coli | - | + | + | + | + | + + | + + | |
| Bacillus pyocyaneus | - | - | + | + + | + + | + + | + + | |
| Micrococcus luteus | - | + | + | + | + + | + + | + + | |
| Enterococcus faecalis | - | + | + + | + + | + + | + + | + + | |
| Aspergillus flavus | - | - | - | - | + + | + + | + + | |
Claims (6)
1, a kind of natural antibacterial composition that extracts from Semen Caesalpiniae Minacis is characterized in that:
Exsiccant Semen Caesalpiniae Minacis was pulverized the 40-200 mesh sieve, added solvent in the ratio of 1-5g/10ml, soaks 1-5 time after, 12-24h at every turn, placing temperature is that 30-40 ℃, rotating speed are to shake 12-24h on the constant temperature shaking table of 180-250r/min, sucking filtration obtains extracting solution; Reclaim the solvent after drying, promptly.
2, the natural antibacterial composition that extracts from Semen Caesalpiniae Minacis as claimed in claim 1 is characterized in that: solvent is water, ethanol or ethyl acetate.
3, a kind of extracting method of the natural antibacterial composition that from Semen Caesalpiniae Minacis, extracts, it is characterized in that: exsiccant Semen Caesalpiniae Minacis was pulverized the 40-200 mesh sieve, ratio in 1-5g/10ml adds solvent, after soaking 1-5 time, each 12-24h, placing temperature is that 30-40 ℃, rotating speed are to shake 12-24h on the constant temperature shaking table of 180-250r/min, and sucking filtration obtains extracting solution; Reclaim the solvent after drying, promptly.
4, the extracting method of the natural antibacterial composition that extracts from Semen Caesalpiniae Minacis as claimed in claim 3 is characterized in that:
When solvent was water, the method that reclaims the solvent after drying was :-80 ℃ of freezing 12-24h vacuum freeze drier dryings.
5, the extracting method of the natural antibacterial composition that extracts from Semen Caesalpiniae Minacis as claimed in claim 3 is characterized in that:
When solvent was ethanol, the method that reclaims the solvent after drying was: 40 ℃ of decompression rotations of alcoholic extract concentrate removes ethanol ,-80 ℃ of freezing 12-24h vacuum freeze drier dryings.
6, the extracting method of the natural antibacterial composition that extracts from Semen Caesalpiniae Minacis as claimed in claim 3 is characterized in that:
When solvent was ethyl acetate, the method that reclaims the solvent after drying is: 45 ℃ of concentrating under reduced pressure of acetic acid ethyl acetate extract were removed ethyl acetate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100776756A CN101019931A (en) | 2007-03-08 | 2007-03-08 | Natural antibiotic composition extracted from crinite seed and its extraction process |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100776756A CN101019931A (en) | 2007-03-08 | 2007-03-08 | Natural antibiotic composition extracted from crinite seed and its extraction process |
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| Publication Number | Publication Date |
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| CN101019931A true CN101019931A (en) | 2007-08-22 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2007100776756A Pending CN101019931A (en) | 2007-03-08 | 2007-03-08 | Natural antibiotic composition extracted from crinite seed and its extraction process |
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| CN (1) | CN101019931A (en) |
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- 2007-03-08 CN CNA2007100776756A patent/CN101019931A/en active Pending
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Open date: 20070822 |