CN101019993A - Natural antibiotic composition extracted from lily and its extraction process - Google Patents
Natural antibiotic composition extracted from lily and its extraction process Download PDFInfo
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- CN101019993A CN101019993A CNA2007100776741A CN200710077674A CN101019993A CN 101019993 A CN101019993 A CN 101019993A CN A2007100776741 A CNA2007100776741 A CN A2007100776741A CN 200710077674 A CN200710077674 A CN 200710077674A CN 101019993 A CN101019993 A CN 101019993A
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Abstract
The present invention discloses one kind of natural antibiotic composition extracted from lily and its extraction process. The extraction process includes crushing dry lily 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, alcohol 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 Bulbus Lilii, 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.
Bulbus Lilii is the dry meat scale leaf of liliaceous plant tiger lily Lilium lancifolium Thunb, Bulbus Lilii lilium brownii F.EBrown var.viriulum Baker or Lilium tenuifolium Lilium pumilulm DC..Excavate autumn, cleans, and strips scale leaf, puts the boiling water part omitted and scald drying.Bulbus Lilii has cough-relieving, eliminates the phlegm, and relievings asthma, and pharmacological actions such as calmness are used to treat diseases such as cough caused by dryness, schizophrenia, pulmonary tuberculosis and climacteric syndrome clinically, has significant curative effect.Contain various trace element, saponin, phospholipid, polysaccharide and alkaloids chemical constituent.But do not see report about the external bacteriostatic activity aspect of Bulbus Lilii,
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 aemginosa), micrococcus luteus (Micrococcus luteus) are had the inhibiting natural antibacterial composition that extracts from Bulbus Lilii.
Another object of the present invention provides the extracting method of the natural antibacterial composition that extracts from Bulbus Lilii.
The natural antibacterial composition that extracts from Bulbus Lilii of the present invention is characterized in that:
Exsiccant Bulbus Lilii 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 Bulbus Lilii, extracts, it is characterized in that: exsiccant Bulbus Lilii 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 Bulbus Lilii, 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 bacillus pyocyaneus, and minimum inhibitory concentration is 12.5mg/ml; To escherichia coli, micrococcus luteus, the minimum inhibitory concentration of Aspergillus flavus is 25mg/ml; To staphylococcus aureus, the minimum inhibitory concentration of enterococcus faecalis is 50mg/ml.The spirit solvent extract is better to micrococcus luteus and aureus with inhibition, and minimum inhibitory concentration is 6.25mg/ml; Minimum inhibitory concentration to escherichia coli and Aspergillus flavus is 12.5mg/ml; Minimum inhibitory concentration to bacillus pyocyaneus and enterococcus faecalis is 25mg/ml; Ethyl acetate solvent has effect preferably to staphylococcus aureus, and minimum inhibitory concentration is 3.13mg/ml; To other for the inhibition effect of examination bacterium slightly a little less than, be 6.25mg/ml to the minimum inhibitory concentration of bacillus pyocyaneus; Minimum inhibitory concentration to Aspergillus flavus is 12.5mg/ml, and to escherichia coli, micrococcus luteus, the minimum inhibitory concentration of enterococcus faecalis are 50mg/ml.Illustrate that the different solvents extract of Bulbus Lilii all has the activity of bacteria growing inhibiting.For the further development and use of Bulbus Lilii 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 Bulbus Lilii 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 Bulbus Lilii 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 50,25,12.5,6.25,3.1,1.5,0.75,0.375mg/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, every gradient scribbles three flat boards, and 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 Bulbus Lilii was pulverized 120 mesh sieves, take by weighing 150g, add 1000ml, 95% ethanol, after soaking 3 times, each 18h, placing temperature is that 30 ℃, rotating speed are to shake 18h on the constant temperature shaking table of 250r/min, sucking filtration obtains extracting solution, 40 ℃ of decompression rotations concentrate and remove ethanol, and-80 ℃ of freezing 12h obtain dry product with the dry 24h of vacuum freeze drier.
Get the extract dry product 0.3g of Bulbus Lilii 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 | ||
| The wine extract | Staphylococcus aureus | - | - | - | - | + | ++ | ++ |
| Escherichia coli | - | - | - | ++ | ++ | ++ | ++ | |
| Bacillus pyocyaneus | - | - | + | + | ++ | ++ | ++ | |
| Micrococcus luteus | - | - | - | - | ++ | ++ | ++ | |
| Enterococcus faecalis | - | - | + | + | ++ | ++ | ++ | |
| Aspergillus flavus | - | - | - | ++ | ++ | ++ | ++ | |
Embodiment 3
Exsiccant Bulbus Lilii 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 Bulbus Lilii 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 50,25,12.5,6.25,3.1,1.5,0.75,0.375mg/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 | ||
| 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 Bulbus Lilii is characterized in that:
Exsiccant Bulbus Lilii 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 Bulbus Lilii 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 Bulbus Lilii, extracts, it is characterized in that: exsiccant Bulbus Lilii 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 Bulbus Lilii 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 Bulbus Lilii 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 Bulbus Lilii 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.
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| CNA2007100776741A CN101019993A (en) | 2007-03-08 | 2007-03-08 | Natural antibiotic composition extracted from lily and its extraction process |
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| CNA2007100776741A CN101019993A (en) | 2007-03-08 | 2007-03-08 | Natural antibiotic composition extracted from lily and its extraction process |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103409988A (en) * | 2013-07-24 | 2013-11-27 | 吴江市七都镇庙港雅迪针织制衣厂 | Wool treating agent containing flos farfarae extract |
| CN103860861A (en) * | 2014-03-06 | 2014-06-18 | 四川农业大学 | Method for extracting lily flavonoid compound |
| CN108310197A (en) * | 2018-02-09 | 2018-07-24 | 海盐县凌特生物科技有限公司 | The preparation method of lily active material |
| CN110195077A (en) * | 2019-06-14 | 2019-09-03 | 长江师范学院 | A method of building Lilium lancifo1ium Thunb efficient genetic trasformation system |
-
2007
- 2007-03-08 CN CNA2007100776741A patent/CN101019993A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103409988A (en) * | 2013-07-24 | 2013-11-27 | 吴江市七都镇庙港雅迪针织制衣厂 | Wool treating agent containing flos farfarae extract |
| CN103409988B (en) * | 2013-07-24 | 2015-06-17 | 苏州爱立方服饰有限公司 | Wool treating agent containing flos farfarae extract |
| CN103860861A (en) * | 2014-03-06 | 2014-06-18 | 四川农业大学 | Method for extracting lily flavonoid compound |
| CN108310197A (en) * | 2018-02-09 | 2018-07-24 | 海盐县凌特生物科技有限公司 | The preparation method of lily active material |
| CN110195077A (en) * | 2019-06-14 | 2019-09-03 | 长江师范学院 | A method of building Lilium lancifo1ium Thunb efficient genetic trasformation system |
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