CN103977028A - Bacteriostatic composition of sweet potato leaf chlorogenic acid and nano zinc oxide - Google Patents
Bacteriostatic composition of sweet potato leaf chlorogenic acid and nano zinc oxide Download PDFInfo
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- CN103977028A CN103977028A CN201410179567.XA CN201410179567A CN103977028A CN 103977028 A CN103977028 A CN 103977028A CN 201410179567 A CN201410179567 A CN 201410179567A CN 103977028 A CN103977028 A CN 103977028A
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- chlorogenic acid
- extracting solution
- zine oxide
- inhibiting composition
- nano zine
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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
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Abstract
The invention discloses a bacteriostatic composition of sweet potato leaf chlorogenic acid and nano zinc oxide, and belongs to the field of traditional Chinese medicine. Combined use of two drugs of a chlorogenic acid extraction liquid and nano zinc oxide in the bacteriostatic composition plays the synergistic effect on inhibition of escherichia coli O157:H7 and staphylococcus aureus. Compared with the prior art, the bacteriostatic composition of the sweet potato leaf chlorogenic acid extract and nano zinc oxide solves the problem of larger drug using amount in a conventional bacteriostatic method, and at the same time, the bacteriostatic composition adopting the two low-dosage bacteriostatic substances decreases the occurrence probability of microbial drug resistance, reduces the toxicity to human bodies, and is safer.
Description
Technical field
The invention belongs to the field of Chinese medicines, relate in particular to a kind of Chinese medicine bateriostatics.
Background technology
Rhizoma Dioscoreae esculentae has another name called Ipomoea batatas Lam., Radix Ipomoeae etc., is most important grain, feedstuff and the raw material of industry in the world, extensively plants in more than 100 countries.Rhizoma Dioscoreae esculentae is integration of edible and medicinal herbs food, has the plurality of health care functions such as invigorating middle warmer and blood, fertile the five internal organs, tonify deficiency gas, physical strength profiting, strengthening the spleen and stomach, kidney tonifying the moon.Also there is Scavenging active oxygen simultaneously, suppress skin aging, suppress cholesterol, prevent emphysema, give protection against cancer and benefit the effects such as cardiovascular health.In Rhizoma Dioscoreae esculentae leaf, except containing above nutritional labeling, be also rich in the compound such as active polysaccharide and chlorogenic acid.Modern pharmacology research shows that chlorogenic acid has antibacterial, antiviral effect.In addition, chlorogenic acid also has the effect such as removing and anti peroxidation of lipid, hepatic cholagogic, antimutagenic and spasmolytic to free radical.Chlorogenic acid, because it has higher medicine and health care function, arouses extensive attention.It is once large with dosage that Chinese medicine preparation concentrated solution is made oral liquid or bacteria inhibiting composition etc., and life-time service affects health.
Summary of the invention
For chlorogenic acid bacteria inhibiting composition ampoule, large and life-time service affects health problem, the inventor, through great many of experiments, finds that nano zine oxide can effectively strengthen chlorogenic acid extracting solution bacteriostasis property, reduces the consumption of chlorogenic acid antibacterial, and adopt two kinds of bacteria inhibiting compositions, more safe.The object of the present invention is to provide a kind of chlorogenic acid nano zine oxide bacteria inhibiting composition antibacterial, reduce the chlorogenic acid antibacterial toxicity to human body with dosage and reduction medicine once.
The invention provides following technical scheme:
Rhizoma Dioscoreae esculentae leaf chlorogenic acid and nano zine oxide bacteria inhibiting composition, contain chlorogenic acid extracting solution, nano zine oxide.
The preparation of described chlorogenic acid extracting solution comprises the steps: often to take Rhizoma Dioscoreae esculentae leaf powder 6g, and adding pH value by solid-liquid ratio 1g:12mL is 4 60% alcoholic solution; Sucking filtration while hot after 80 DEG C of reflux, extract, 2.5h of constant temperature water bath, reflux, extract, 1h after filtering residue repeats 1 time, sucking filtration, merging filtrate.Divide and add for 2-3 time active carbon to decolour in the ratio of 2% left and right, sucking filtration after the 30min that at every turn decolours, filtrate is centrifugal 30min under 5000r/min condition, collects supernatant for subsequent use;
Described nano zine oxide is the nano zine oxide colloid solution of 10mg/mL.
Suppress Escherichia coli O 157: when H7, described chlorogenic acid extracting solution Content of Chlorogenic Acid concentration 47.8 μ g/mL, nano oxidized zinc concentration is 10 mg/mL, volume ratio 70:3.
While suppressing staphylococcus aureus, described chlorogenic acid extracting solution Content of Chlorogenic Acid concentration 47.8 μ g/mL, nano oxidized zinc concentration is 10mg/mL, volume ratio 25:1.
The invention still further relates to, described bacteria inhibiting composition suppresses Escherichia coli O 157 in preparation: the application in H7 medicine.
Bacteria inhibiting composition suppresses the application in staphylococcus aureus medicine in preparation.
The invention has the beneficial effects as follows: the application of Nano-Zinc of the present invention can effectively strengthen chlorogenic acid bacteriostasis property, increase the dissolubility of nano zine oxide, reduce the consumption of chlorogenic acid antibacterial, more unexpected is that share of two kinds of medicines played synergism to suppressing Escherichia coli O 157: H7 and staphylococcus aureus bacterium, this bacteria inhibiting composition adopts the antibacterial substance of two kinds of low dosages simultaneously, with respect to the chlorogenic acid or the nano zine oxide that use high dose, more safe, can be used as external coating agent etc. safely and reliably.
Detailed description of the invention
Embodiment 1
1, the preparation of chlorogenic acid extracting solution comprises the steps: accurately to take Rhizoma Dioscoreae esculentae leaf powder 6g, and adding pH value by solid-liquid ratio 1g:12mL is 4 60% alcoholic solution; Sucking filtration while hot after 80 DEG C of reflux, extract, 2.5h of constant temperature water bath, reflux, extract, 1h after filtering residue repeats 1 time, sucking filtration, merging filtrate.Divide and add for 2-3 time active carbon to decolour in the ratio of 2% left and right, sucking filtration after the 30min that at every turn decolours, filtrate is centrifugal 30min under 5000r/min condition, collects supernatant for subsequent use.The extracting solution of gained carries out standard quantitative through ultraviolet-visible photometer to chlorogenic acid.The content of extracting solution Content of Chlorogenic Acid is 47.8 μ g/mL.
2, nano oxidized zinc solution is prepared according to following method: the nano zine oxide 0.01g that gets the production of Shanghai Aladdin company limited is dissolved in the distilled water of 1mL, and concussion mixes the nano zine oxide colloid solution that obtains 10mg/mL, and particle diameter is 200nm.
3, the collaborative bacteriostasis of chlorogenic acid extracting solution and nano zine oxide.
The cultivation of a antibacterial
Single bacterium colony of the Escherichia coli O 157 on picking LB agar culture medium is in 10mL LB broth bouillon, be placed in 37 DEG C of incubators and cultivate 12 h, be inoculated in the LB broth bouillon of 5 mL by 1% inoculum concentration again, cultivate after 5h for 37 DEG C, get 2 gradients of 1 mL above-mentioned bacterium liquid 1xPBS water serial dilution, final bacterium liquid is approximately 10
6cFU/mL.
B chlorogenic acid and the nano zine oxide synergism on bacteriostasis property
Get respectively the above-mentioned bacterium liquid having diluted of 1 mL in the centrifuge tube of 1.5 mL sterilizings, to adding respectively 70 μ L chlorogenic acid extracting solution in No. 1 centrifuge tube, (content of chlorogenic acid element is 47.8 μ g/mL, lower same), to the nano zine oxide that adds 3 μ L concentration 10 mg/mL in No. 2 centrifuge tubes, be the nano zine oxide of 10 mg/mL to adding 35 μ L chlorogenic acid extracting solution and 1.5 μ L concentration in No. 3 centrifuge tubes, do not add any above-mentioned antibacterial.For ensureing that every pipe liquid final volume equates, 1xPBS water polishing for residual quantity.Be placed in 37 DEG C and cultivate 2 h.Test parallel three times for every group.
C plate count
By each group of 4 gradients of 1xPBS serial dilution for liquid to be measured, 10
-2, 10
-3, 10
-4each middle taking-up 200 μ L are on LB flat board.After flat board dries up, be inverted and cultivate 12h at 37 DEG C, count after growing bacterium colony, taking 30-300 colony-forming units (CFU) as effective count range.Every milliliter of original bacteria liquid viable count=plate count * extension rate * 6
Experimental result is as follows:
| Antibacterial | Add antibacterial rear the clump count can number going out |
| The nano zine oxide of 3 μ L concentration 10 mg/mL | 1360 CFU/mL |
| 70 μ L chlorogenic acid extracting solution (content of chlorogenic acid element is 47.8 μ g/mL) | 698 CFU/mL |
| 35 μ L chlorogenic acid extracting solution (content of chlorogenic acid element is 47.8 μ g/mL) and 1.5 μ L concentration are the nano zine oxide of 10 mg/mL | 0 CFU/mL |
Experiment shows: bacteria inhibiting composition provided by the invention not only has the nano zine oxide of promotion dissolution, simultaneously by reducing the Mlc of chlorogenic acid, greatly reduce the problem of the disposable consumption of chlorogenic acid antibacterial, and share of two kinds of antibacterial substances played collaborative effect to suppressing Escherichia coli O 157: H7.
Embodiment 2
1, the preparation process of chlorogenic acid extracting solution: with embodiment 1
2, the preparation process of nano zine oxide: with embodiment 1
3, chlorogenic acid extracting solution and nano zine oxide are worked in coordination with bacteriostasis
The cultivation of a antibacterial
Single bacterium colony of staphylococcus aureus on picking LB agar culture medium is in 10mL LB broth bouillon, be placed in 37 DEG C of incubators and cultivate 12 h, be inoculated in the LB broth bouillon of 5 mL by 1% inoculum concentration again, cultivate after 5h for 37 DEG C, get 2 gradients of 1 mL above-mentioned bacterium liquid 1xPBS water serial dilution, final bacterium liquid is approximately 10
6cFU/mL.
B chlorogenic acid and the nano zine oxide synergism on bacteriostasis property
Get respectively the above-mentioned bacterium liquid having diluted of 1 mL in the centrifuge tube of 1.5 mL sterilizings, to adding respectively 50 μ L chlorogenic acid extracting solution in No. 1 centrifuge tube, (content of chlorogenic acid element is 47.8 μ g/mL, lower same), to the nano zine oxide that adds 2 μ L concentration 10 mg/mL in No. 2 centrifuge tubes, be the nano zine oxide of 10 mg/mL to adding 25 μ L chlorogenic acid extracting solution and 1 μ L concentration in No. 3 centrifuge tubes, do not add any above-mentioned antibacterial.For ensureing that every pipe liquid final volume equates, 1xPBS water polishing for residual quantity.Be placed in 37 DEG C and cultivate 2 h.Test parallel three times for every group.
C plate count
By each group of 4 gradients of 1xPBS serial dilution for liquid to be measured, from 10
-2, 10
-3, 10
-4each middle taking-up 200 μ L are on LB flat board.After flat board dries up, be inverted and cultivate 12h at 37 DEG C, count after growing bacterium colony, taking 30-300 colony-forming units (CFU) as effective count range.Every milliliter of original bacteria liquid viable count=plate count * extension rate * 6
Experimental result is as follows:
| Antibacterial | Add antibacterial rear the clump count can number going out |
| The nano zine oxide of 2 μ L concentration 10 mg/mL | 1450 CFU/mL |
| 50 μ L chlorogenic acid extracting solution (content of chlorogenic acid element is 47.8 μ g/mL) | 562 CFU/mL |
| 25 μ L chlorogenic acid extracting solution (content of chlorogenic acid element is 47.8 μ g/mL) and 1 μ L concentration are the nano zine oxide of 10 mg/mL | 0 CFU/mL |
Experiment shows: bacteria inhibiting composition provided by the invention not only has the nano zine oxide of promotion dissolution, simultaneously by reducing the Mlc of chlorogenic acid, greatly reduce the problem of the disposable consumption of chlorogenic acid antibacterial, and share of two kinds of antibacterial substances played collaborative effect to suppressing staphylococcus aureus.
Claims (6)
1. Rhizoma Dioscoreae esculentae leaf chlorogenic acid and nano zine oxide bacteria inhibiting composition, is characterized in that containing chlorogenic acid extracting solution, nano zine oxide;
Bacteria inhibiting composition as claimed in claim 1, is characterized in that the preparation of described chlorogenic acid extracting solution comprises the steps: often to take Rhizoma Dioscoreae esculentae leaf powder 6g, and adding pH value by solid-liquid ratio 1g:12mL is 4 60% alcoholic solution; Sucking filtration while hot after 80 DEG C of reflux, extract, 2.5h of constant temperature water bath, reflux, extract, 1h after filtering residue repeats 1 time, sucking filtration, merging filtrate.
2. divide and add for 2-3 time active carbon to decolour in the ratio of 2% left and right, sucking filtration after each decolouring 30min, filtrate is centrifugal 30min under 5000r/min condition, collects supernatant for subsequent use, and the extracting solution of gained carries out standard quantitative through ultra-violet and visible spectrophotometer to chlorogenic acid; Described nano zine oxide is the nano zine oxide colloid solution of 10mg/mL.
3. bacteria inhibiting composition as claimed in claim 1, is characterized in that suppressing Escherichia coli O 157: when H7, and described chlorogenic acid extracting solution Content of Chlorogenic Acid concentration 47.8 μ g/mL, nano oxidized zinc concentration is 10 mg/mL, volume ratio 70:3.
4. bacteria inhibiting composition as claimed in claim 1, while it is characterized in that suppressing staphylococcus aureus, described chlorogenic acid extracting solution Content of Chlorogenic Acid concentration 47.8 μ g/mL, nano oxidized zinc concentration is 10mg/mL, volume ratio 25:1.
5. as described in as arbitrary in claim 1 ~ 4, bacteria inhibiting composition suppresses Escherichia coli O 157 in preparation: the application in H7 medicine.
6. as described in as arbitrary in claim 1 ~ 4, bacteria inhibiting composition suppresses the application in staphylococcus aureus medicine in preparation.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113371753A (en) * | 2021-06-07 | 2021-09-10 | 江苏农林职业技术学院 | Nano zinc and green synthesis method and application thereof |
-
2014
- 2014-04-30 CN CN201410179567.XA patent/CN103977028A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| FATEMEH BARZEGARI FIROUZABADI ETAL: "ZnO nanoparticle suspensions containing citric acid as antimicrobial to control listeria monocytogenes, escherichia coli, staphylococcus auresu and bacillus cereus in mango juice", 《FOOD CONTROL》 * |
| 李瑞国等: "甘薯绿原酸的提取及抑菌作用", 《江苏农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113371753A (en) * | 2021-06-07 | 2021-09-10 | 江苏农林职业技术学院 | Nano zinc and green synthesis method and application thereof |
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