CN102978241A - Method for producing metal nanoparticles - Google Patents
Method for producing metal nanoparticles Download PDFInfo
- Publication number
- CN102978241A CN102978241A CN2012104486448A CN201210448644A CN102978241A CN 102978241 A CN102978241 A CN 102978241A CN 2012104486448 A CN2012104486448 A CN 2012104486448A CN 201210448644 A CN201210448644 A CN 201210448644A CN 102978241 A CN102978241 A CN 102978241A
- Authority
- CN
- China
- Prior art keywords
- silver
- nanometer
- gold
- bacterium
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- 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
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Manufacture Of Metal Powder And Suspensions Thereof (AREA)
- Powder Metallurgy (AREA)
Abstract
This invention provides a method for producing a composition comprising colloidal nanoparticles of metals including silver, gold, zinc, mercury, copper, palladium, platinum, or bismuth, by contacting a metal or metal compound with bacteria. An embodiment of the method comprises a step of incubating probiotic bacteria with an aqueous solution comprising at least 4 mM of a silver or gold salt. A resulting nanosilver-containing composition is useful as a highly efficient antimicrobial agent, for instance when impregnated onto a carrier, or an algicide agent or a herbicide agent.
Description
The application is International Application Serial No. PCT/EP2007/006145, international filing date on July 5th, 2007, China national phase application numbers 200780025198.7, the dividing an application of the application for a patent for invention of title " method for preparing metal nanoparticle ".
Invention field
The present invention relates at bacterial film preparation colloidal metal compound.The present invention also relates to prepare by bioprocess the method for silver or gold nano grain.Concrete, the present invention relates to use probiotic bacterium, such as Bacterium lacticum (Lactobacillus), produce under given conditions the metal nano precipitation, especially silver or gold nano grain are to improve its antimicrobial efficient.The invention still further relates to a kind of sterilized product that comprises carrier, this carrier is that usefulness contains the colloidal nano silver of described method preparation or the composition of nanometer gold floods.
Background of invention
Comprise in a large number the microbiological contamination material in processing, such as water, particularly family or industrial circulating water, and need effective sterilizing process in sewage (such as the sewage that produces in the food-processing industry) process, because health, operation or environment reason, these sewage can not discharge or reuse before being untreated.When processing buildings for example, equipment, container, air-conditioning system surperficial, also need effective sterilizing process.With the sterilizing process of environmentally compatible mainly based on reactive oxygen compounds, such as hydrogen peroxide or quaternary ammonium compound monomer.
A kind of active suitable mild antibiotic agent with sterilization idiocratic during hydrogen peroxide.Can suppress the growth of some bacteriums when known in which hydrogen peroxide concentration is 25mg/l, yet even if under the hydrogen peroxide effect of greater concn, effective minimizing of microorganism count also needs a few hours or also needs uv irradiating.Need expensive equipment and a large amount of electric consumptions and produce ultraviolet ray.Therefore when a large amount of polluting material such as water are carried out disinfection, for example sewage work disposes of sewage and when discharging, this class methods shortage feasibility and/or uneconomical.Therefore, different methods has been attempted to overcome these inferior positions in this area.
Silver ions known in the art or have height toxicity for microorganism based on the compound of silver, thereby many common bacteria kinds are comprised that intestinal bacteria have shown potent anti-microbial effect.Shown that silver nano-grain and the hyperbranched macromolecular crossbred of amphiphilic have effective antimicrobial surface coating.The stable aqueous dispersion of finding the silver nano-grain of nontoxic silver element water-sol form has strong germicidal action to intestinal bacteria, and 50 μ g/cm3 concentration can 100% bacteria growing inhibiting.The discovery silver nano-grain can be accumulated in bacterial film, has an effect with some structural element of bacterial film in some way, therefore causes its structural changes, degraded, finally causes necrocytosis.Report that because excessive carboxyl and other group dissociates in the film, bacterium surface is electronegative generally under biology pH value.Show, make its surface charging because silver nano-grain motion and the friction in the substrate of film carbon advanced in embedding, so perhaps electrostatic force be the reason of nano particle and bacterium interaction.In addition, silver is tending towards having higher avidity, with the compound reaction that contains p and s among bacterial film and the DNA.The third possible binding mode is the silver ions that discharges the bactericidal effect that may produce silver nano-grain.
Reported several microorganisms, for example Bacterium lacticum, fungi Fusarium oxysporum (Fusarium oxysporum) can arrive its cell surface with Ag (I) biological adsorption, and be used for simultaneously being reduced into Ag (0) by reductase enzyme effect or electron shuttle quinone or both, thereby this ion is detoxified.
A kind of silver nano-grain that comprises magnitude range 1-100nm bio-stable known in the art, and the anti-microbial agents that contains the no cytotoxicity of the nano particle of described bio-stable and the carrier that concentration is 1-6ppm.
Also known a kind of method for preparing colloidal silver-biomolecules mixture, the method comprises:
-mixture of biomolecules, silver salt and halogen ion source is provided in single solution; And
-with this mixture of rayed of visible region wavelength, wherein silver salt and halogen ion source are water miscible; The content of biomolecules, silver salt, halogen ion source can make irradiating step obtain colloidal silver-biomolecules mixture.
A kind of method for preparing the colloidal metal particle of nanosized is disclosed, described method is included in 15-40 ℃ the temperature range with the wet fungi of the solution-treated of metal ion or fungus extract 2-120 hour, and the separating bio material is to obtain the colloidal metal particle of nano-scale.
The production method of conventional preparation silver nano-grain has a lot of inferior positions, such as high production cost, the byproduct that produces remarkable ratio or the upper limit that concentrations of nanoparticles is obtained in existence.For example, rear a kind of production method needs the very long production time and utilizes that the possible fungi of causing a disease is arranged.Therefore this area needs a kind of reliable, cheap and method for preparing silver nano-grain of reducing or avoiding byproduct to produce.
Studied, the Ag of Bacterium lacticum (I) biological adsorption process, this process pH dependency, the temperature dependency in temperature range 10-60 ℃ and Bacterium lacticum in pH scope 2-6 is with Ag
+Be reduced to Ag
0Mechanism.
This area is also known a kind ofly to prepare the method for silver nano-grain by biological reducing, namely utilizes Aeromonas (Aeromonas sp.) and silver ions, ammonia and sodium hydroxide mixed, 60 ℃ of stoichiometric numbers hour.
The defective of aforesaid method is to need high temperature, acid pH and/or long incubation time, or not enough by the fungicidal activity of the silver nano-grain of its generation.
Therefore this area need to prepare by the method that does not have these inferior positions silver or gold nano grain.
The method that this area also needs is simple, eco-friendly, repeatably preparation has silver or the gold nano grain of high antimicrobial property.
This area also needs to prepare the known gold of some medical use or the correlation method of silver nano-grain of can be used for.
This area is also understood, and the colloidal form of the metal except gold or silver and the compound of described metal has valuable characteristic and application.For example, colloidal bismuth subcitrate is water-soluble, and especially when the pH scope was 3-8, in decades, it and Antibiotic combination were used for the treatment of gastric duodenal ulcer and helicobacter pylori infection.The mercury of external application colloidal form, inorganic mercury compound and mercury metal salve have various treatments to be used, and comprises the infectious eczema for the treatment of or pustulosis (mercury salt), treatment syphilis (calomel), treatment psoriasis (red precipitate or ammonification mercury).Use the colloidal form catalysis number of chemical reaction of palladium and platinum, comprise organic reducing and hydrogenolysis etc.The Pt nanoparticle of colloidal form is also as anticarcinogen.Optional electrocuprol with the Whitfield's ointment chelating is potent antiphlogistic, and hypogloeeis (administration) form of known electrocuprol or colloid zinc can be resisted flu and influenza.In addition, colloid zinc is especially effective at anti-virus aspect.In all these different fields, need to provide for a long time multi-form colloidal metal or colloidal metal compound, in order to improve it in the effect in related application field.
Summary of the invention
In a broad sense, the present invention relates to use bacterium at bacterial film preparation colloidal metal compound, and coated bacterium is as the subsequent applications of biocide.The present invention relates to specifically:
By under controlled pH, the mixture of described bacterium with metal-salt or other salt being contacted, at bacterial film preparation colloidal metal compound, in order to use bacterium to prepare the colloidal metal compound, and
Use the bacterium of coated metallic compound on the above-mentioned film as biocide.
In one embodiment, the present invention relates to utilize probiotic bacterium and other bacterium to prepare the metal nano precipitation, described metal nano precipitation can be used as the biocide of tap water, topcoating and other material.
More particularly, some bacteriums can be reduced to Ag (I) salt and be deposited in the colloid Ag (0) that cell surface forms the nanometer Ag particle.The coated biological substance of colloidal silver or other metal nano precipitation is easy to by filtering or centrifugally collecting from water, and can wash, rinsing and other processing, and form the colloid product with strong antimicrobial property in (dilution) suspension or when being processed as coating.
What is interesting is, multiple probiotic bacterium, namely bacterium industrial, useful HUMAN HEALTH when coming across people's digestive tube has shown that it produces the ability of Ag nanometer precipitation at cell surface.This bacterioid includes but not limited to: benefit is given birth to type lactobacillus fermentum (Lactobacillus fermentum) bacterial strain.
By in concentrated bacterial cell culture, adding the combination (AgNO of specific salts
3, NH
4Cl, NaOH etc.) and control pH, form the colloidal silver product with strong antimicrobial property, other metal-salt and the combination of some bacterial isolates are obtained having the nanometer precipitation of similar characteristics, this also is a part of the present invention.
By regulating " silver material " and " biological cellular material " ratio (Ag:CDW, wherein CDW=dry cell weight), the reactivity of final colloidal silver product may be different with characteristic, and this relates to the colloid granularity, colloidal solid distributes and other characteristic.
The colloidal silver compound that bacterium surface produces has very widely to be used, include but not limited to: water sterilization, as the sterilizing agent in the cleaning product, as sanitising agent, be mixed with antimicrobial coating, medical use, human consumption product, be used for textiles, salve and lubricant, be used as catalyzer etc.
Production process is simple and clear, cost economy, output high, be easy to mass-producing, the size of particle and distribute controlled, the antimicrobial acivity of the nanometer silver that produces under extremely low (ppb) concentration be better than other colloidal silver product.And this product can be processed into multi-form: dried forms, suspension form or " wetting " agglomerate can be mixed with different application.Because available pure water rinsing and non-loss of activity be not so chemical agent residue occurs in the final product.
The use probiotic bacterium has been started the many application in health care and the foodstuffs industry.The bacterium product of coated Ag especially is fit to following application:
Be mixed with sterilization cleaning product (hospital, laboratory, animal production site etc.)
The porcelain filter and other strainer that are used for water sterilization comprise tap water, swimming-pool water, animal water of productive use, water industry raising water etc.
Be used for sterilization coating: polymkeric substance, textile fibres and metal
Be fit to be mixed with salve, the lubricant of sterilization skin
Be used for the sterilization of tap water: developing country, knapsack family, aircraft and many other sides (simple and easy method) (easy-drop method), and
Enantiopathy substance: legionella (Legionella), Cryptosporidium (Cryptosporidium), hepatitis virus (Hepatitis), simplexvirus (Herpes), pseudomonas (Pseudomonas), staphylococcus (Staphylococcus), dissimilar bacteriums, fungi and virus.
An object of the present invention is to provide high-quality gold or silver nano-grain.First aspect of the present invention provides the biological method that improved preparation comprises the composition of colloidal silver or gold nano grain, described method comprises uses probiotic bacterium, especially Lactobacillus species, such as lactobacillus fermentum, and the aqueous solution of described biological substance with silver (I) salt or gold (III) salt contacted.The present invention is based on beyond thought discovery: some specific procedure parameter that biological reducing prepares silver or gold nano grain greatly affects production efficiency and the attribute that obtains nano particle.Ad hoc approach of the present invention greatly affects the antimicrobial acivity of gained silver-containing nanoparticles composition specifically.
Another aspect of the present invention is that the silver or the gold nano grain composition that obtain by biological reducing under these actual conditionses also can be processed, and keeps as separating from biological substance even improves its active or other correlation properties, such as stability in storage.In addition, by oxidizing substance such as superoxide or peracid salt the silver that obtains by biological reducing under these actual conditionses or gold nano grain composition are carried out the later stage chemical treatment even can strengthen the characteristic of gained Nanoparticulate compositions.
Size and the distribution that to also have an advantage be gained silver or gold nano grain of process of the present invention repeat controlled.
Another advantage of the present invention is described method by reducing having the demand of genotoxic potential and/or expensive chemical, within the obviously short time, with low-cost and eco-friendly mode, obtains highly believable result.The biological substance that uses of the present invention comes from harmless microorganism to a great extent, such as probiotic bacterium, thereby residual without Harmful chemicals in the composition that obtains of the inventive method.Therefore an advantage of the present invention is that the method provides a kind of composition, and said composition does not affect this type of organism substantially after being applied to the eucaryon organism.In an embodiment, the invention provides the composition with high antimicrobial acivity, said composition equally can anti-ocean pathogenic agent, and does not substantially affect the eucaryon organism.Another advantage of the present invention is, the present invention can prepare the composition that contains highly concentrated nano silver or nanometer gold, and the nanometer silver that comprises or nanometer gold are comprised of its metallic state separately substantially, for example, minute other, comprise in whole silver-colored compositions and surpass about 95% Ag
0Or whole golden compositions comprise the Au above about 95%
0
Another advantage of the present invention is, the processing that products therefrom or composition can be simple and safe and maintenance even improve its activity.Composition can be dried, or remains suspended state or wet agglomerate, can be prepared to multi-form preparation, such as aerosol preparations or be impregnated on the carrier, and because the stability of nano-Ag particles can not affect its antimicrobial acivity.
In another embodiment, the present invention relates to use the colloidal silver composition of according to the method described above preparation as algicide or weedicide.
Definition
According to the object of the invention, term used herein " nanometer silver " or " nanometer Ag " refer to argent (Ag
0) nano particle.Within the implication of the present invention, described nano particle can or can not be deposited on the biological substance.These nanoparticle size range are the about 100nm of about 0.1nm-, and for example scope is the about 5nm of about 0.5nm-.These nano particle sizes distribute near its mean sizes.
According to the object of the invention, term used herein " nanometer gold " or " nanometer-Au " refer to metallic gold (Au
0) nano particle.Within the implication of the present invention, described nano particle can or can not be deposited on the biological substance.These nanoparticle size range are the about 100nm of about 0.1nm-, and for example scope is the about 5nm of about 0.5nm-.
According to the object of the invention, term used herein " biological substance " refers to comprise or originate from the organic materials for the preparation of the bacterial species of " nanometer silver " or " nanometer gold ".
According to the object of the invention, term used herein " probiotic bacterium " refers to bacterium, and this bacterium can produce useful effect to described host health when giving q.s to host such as Mammals, marine animal (such as fish) or people.
According to the object of the invention, term used herein " silver (I) " or " Ag (I) " refer to silver ions or the Ag of unit price positively charged
+
According to the object of the invention, term used herein " gold (I) " or " gold (III) " refer to respectively unit price and trivalent positively charged gold ion.
Brief Description Of Drawings
Fig. 1 demonstration utilizes the processing of different concns nano-Ag particles to the antimicrobial effect of total cell count and intestinal bacteria survival according to embodiment of the present invention.
Fig. 2 shows the X-ray diffraction analysis chart according to the embodiment of the present invention nano-Ag particles.
Fig. 3 shows according to embodiment of the present invention silver and the impact of dry cell weight ratio on the anti-microbial activity of described particle antagonism Salmonella typhimurtum (Salmonella typhimurium) in the process of preparation nano-Ag particles.
Fig. 4 shows the X-ray diffraction analysis chart according to another embodiment of the present invention nano-Ag particles.
Detailed Description Of The Invention
First aspect of the present invention provides a kind of simple method for preparing the composition that comprises colloidal nano silver or nanometer gold, and the method comprises the step that probiotic bacterium is hatched with the aqueous solution that comprises the silver of 4mM at least or golden salt.
According to the present invention, suitable probiotic bacterium includes but not limited to Bacterium lacticum, bifidus bacillus, intestinal bacteria, faecalis, yeast and bacillus.Probiotic bacterium can belong to but be not limited to following kind: lactobacillus sake (Lactobacillussakei), Lactobacterium acidophilum (Lactobacillus acidophilus), cheese Bacterium lacticum (Lactobacilluscasei), lactobacillus crispatus (Lactobacillus cripatus), bulgaricus ccm (Lactobacillusdelbrueckii subspecies bulgaricus), lactobacillus fermentum (Lactobacillus fermentum), lactobacillus gasseri (Lactobacillus gasseri), Lactobacillus johnsonii (Lactobacillus johnsonii), lactobacillus paraceasi (Lactobacillus paracasei), plant lactobacillus (Lactobacillus plantarum), lactobacillus reuteri (Lactobacillus reuteri), lactobacillus rhamnosus (Lactobacillus rhamnosus), bifidobacterium (Bifidobacterium bifidum), short bifidus bacillus (Bifidobacterium breve), bifidobacterium infantis (Bifidobacterium infantis), long bifidus bacillus (Bifidobacterium longum), lactic acid bifidus bacillus (Bifidobacterium lactis), bifidobacterium adolescentis (Bifidobacteriumadolescentis), intestinal bacteria (Escherichia coli) Nissle, yeast probiotic bacterium (Saccharomycesboulardii), thermophilus streptococcus (Streptococcus thermophilus), faecium (Enterococcusfaecium), Bacillus licheniformis (Bacillus licheniformis), bacillus cereus (Bacilluscereus), subtilis (Bacillus subtilis), bacillus megaterium (Bacillusmegaterium), have a liking for sour genus bacillus (Bacillus acidophilus), short and small genus bacillus (Bacilluspumilus), multiple ferment genus bacillus (Bacillus polyfermenticus), Bacillus clausii (Bacillus clausii), bacillus laterosporus (Bacillus laterosporus), bacillus putrificus (Bacillussporogenes), Bacillus coagulans (Bacillus coagulas) and poly-viscosity bacillus (Bacillus polymyxa).
According to the purpose of the different embodiments of the inventive method, can use any water soluble silver salt.Term used herein " silver salt " also comprises hydrate and other solvate of this type of silver salt.Usually, water soluble silver salt defined herein refers to, under the temperature of the inventive method operation, and under room temperature, the water-soluble silver salt that is at least 0.1g/L.Described silver salt includes but not limited to inorganic silver salt or organic silver salts, as but be not limited to Silver monoacetate, silver chloride, silver perchlorate, silver chlorate, Silver monobromide, silver fluoride, silver lactate, Silver Nitrate, Sulfuric acid disilver salt or silver tartrate.
The purpose of different embodiments can be used any water-soluble golden salt according to the present invention.Term used herein " golden salt " also comprises hydrate and other solvate of this eka-gold salt.Usually, water-soluble golden salt defined herein refers to, under the temperature of the inventive method operation, and under room temperature, the water-soluble golden salt that is at least 0.1g/L.Gold salt is not limited to unit price or trivalent.Gold salt is not limited to inorganic golden salt or organic golden salt or golden salt mixture and can is, such as but not limited to gold trichloride (III), a hydration disodium aurothiomalate, gold tribromide (III), gold triiodide (III) and nitric acid gold (III).
According to an embodiment of the invention, silver or the initial concentration of golden salt in the aqueous solution of hatching with it is 4mM at least, 10mM at least for example, or as a concrete example be at least 50mM.
According to another embodiment of the present invention, the described aqueous solution also can comprise other easy component that affects behavior, and these behaviors especially improve the resulting composition characteristic.For this, need in the present invention in a kind of embodiment of nanometer silver, the method can comprise that this aqueous solution also can comprise ammonia or ammonium salt with probiotic bacterium (such as this paper aforementioned definitions) and the step that contains the aqueous solution of 4mM silver salt at least and hatch.Be fit to the ammonium salt of this embodiment such as but not limited to ammonium chloride, ammonium nitrate, ammonium phosphate, ammonium sulfate, volatile salt, ammonium formiate and brometo de amonio.The ammonia that uses in this embodiment of the present invention and/or the amount of ammonium salt preferably are enough to form a large amount of silver-ammonia or silver-ammonium mixture, such as but not limited to shape such as Ag (NH
2)
+And/or { Ag (NH
3)
2}
+Silver (I)-ammonia mixture.This embodiment variant according to the present invention, the aqueous solution of hatching with it also can comprise the alkali metal hydroxide of appropriate amount, such as but not limited to sodium hydroxide or potassium hydroxide.This type of appropriate amount may be defined as the pH scope that reaches suitable, as hereinafter releasing.
According to another embodiment of the present invention, described method comprises the step that probiotic bacterium (such as this paper aforementioned definitions) and the aqueous solution that contains the gold of 4mM at least salt are hatched, this aqueous solution also can comprise the alkali metal hydroxide of appropriate amount, such as but not limited to the sodium hydroxide when not having ammonia and/or ammonium salt or potassium hydroxide.
Suitable alkali metal hydroxide can be added into and hatch in the aqueous solution such as but not limited to sodium hydroxide or potassium hydroxide, and is high to about 1M to concentration.Preferably, be at least at pH under 8 the condition and hatch, for example in the scope of about 8-about 12, or in a more specific embodiment scope from about 8.5-about 11.
According to an embodiment of the invention, the ratio of silver-colored or golden weight and probiotic cell dry weight (hereinafter being abbreviated as CDW) is at least about 0.01, for example is at least about 0.05 or be at least about 0.1.According to another embodiment of the present invention, it is about 20 that Ag:CDW or Au/CDW weight ratio are no more than, and preferably is lower than 10, for example is lower than 5.
According to an embodiment of the invention, the incubation step of described method is carried out under about 5 ℃-Yue 45 ℃ condition, and preferred about 15 ℃-Yue 35 ℃, room temperature for example.
As another embodiment of the present invention, the incubation step of the method can be carried out in about 1 second-30 minutes time range, for example about 5 seconds-Yue 20 minutes.The Best Times that those of skill in the art can utilize limited measuring to hatch, this depends on other method parameter, such as but not limited to, whether silver-colored or golden salt concn, incubation temperature, Ag:CDW or Au/CDW weight ratio, ammonia or ammonium salt occur etc.As known in the art, hatch under the stirring condition at the part incubation time at least.
Method of the present invention also comprises the resulting step that contains the composition of colloidal silver or gold nano grain of further processing.Described further processing can comprise one or more steps, such as but not limited to, from silver or gold nano grain, be removed to the small part biological substance, or biological substance separated such as ultrasonic method by machinery, zymetology and/or physics and chemistry processing.Arbitrarily the method for this type of removal or separating bio material is known to the those skilled in the art.In addition or in addition, described processing also can comprise chemical treatment step, this step is in order to stable or improve some desired characteristic that gained contains colloidal silver or gold nano grain composition.As this type of chemically treated embodiment, the gold of gained or silver nanoparticle composition can hatched aftertreatment according to the present invention, optional with the oxidizing substances such as superoxide or peracid salt removal biological substance, have the stability of raising and/or silver or the gold nano grain precipitation of (comparing with nanometer silver) higher antimicrobial acivity with generation.In the embodiments of the present invention scope, suitable organic and inorganic peroxide includes but not limited to: hydrogen peroxide, Peracetic Acid etc.The suitable peracid salt that can use in this embodiment of the present invention includes but not limited to: the alkaline water soluble of the rear formation hydrogen peroxide that can dissociate for example, when this type of salt is dissolved in water, discharges peroxide ion.Suitable example comprises percarbonate, perborate, peroxide silicate and the superphosphate of being combined with positively charged ion such as basic metal.Especially preferred is the formula 2Na that sees service
2CO
3﹒ 3H
2O
2Sodium carbonate peroxide.In order to support this embodiment of the present invention, those of skill in the art understand:
-on disinfecting power, this type of peracid salt is better than hydrogen peroxide,
-hydrogen peroxide is weak sterilizing agent and poor to bacterial penetration power, and
-water-soluble and when discharging hydrogen peroxide when the peroxide hydrochlorate, basic metal is captured a proton from the hydrogen peroxide that discharges and is formed the hydroperoxide ion, compares with hydrogen peroxide, and this peroxide ion is strong sterilizing agent and is easy to penetrate into bacterium.
In the random time of process of the present invention, utilize any those skilled in the art institute perception method the solids component that contains colloidal nano silver or nanometer gold can be separated with liquid component.For example, can from liquid ingredient, separate by centrifugal rear decant or by filtering.
In the method for the invention, by the one or more operation conditions of careful adjustment, such as but not limited to pH, incubation temperature, salt type and salt concn at least part of gold or silver salt are replaced with mantoquita.In the method for the invention, by the one or more operation conditions of careful adjustment, such as but not limited to pH, incubation temperature, salt type and (gold or silver) salt concn at least part of probiotic bacterium kind is replaced with alternative microorganism or bacterium.This type of alternative bacterium can be selected from it is generally acknowledged environmentally safe bacterium, more specifically those known bacteriums with biological reducing ability.
Although method of the present invention is mainly described with reference to silver or gold in this article, but do not limit its statement with broad sense and be applied to other metal or metallic compound, as long as to one or more operation conditions, such as but not limited to pH, incubation temperature, salt type and salt concn, make appropriate change.This type of change is in those of skill in the art's the normal experiment scope, supposes that conventional instruction is included into this paper.In the scope of the invention cherish a special interest metal comprise zinc, mercury, copper, palladium, platinum and bismuth.
Based on beyond thought discovery: the effective concentration of this type of nanometer silver composition is extremely low in bioguard treatment, depend on target bacteria, 0.5ppm or lower according to appointment, 0.05ppm or lower for example, and another discovery: in finite time, as observed a large amount of minimizings of undesired bacteria amount in no more than 5 hours, second aspect of the present invention is the antimicrobial purposes of the nanometer silver composition of aforesaid method preparation.The present invention's suitable bacteria target spot in this respect comprises most gram-positives and gram-negative bacteria, such as but not limited to, Pseudomonas aeruginosa (Pseudomonas aeruginosa) (such as the CMCM-2-22 bacterial strain), pseudomonas cepacia (Pseudomonas cepacia), enterobacter cloacae (Enterobacter cloacae), enterobacter agglomerans (Enterobacter agglomerans), Klebsiella Pneumoniae (Klebsiella pneumoniae) (such as the ATCC-10031 bacterial strain), intestinal bacteria (Eschericia coli), streptococcus faecium (Streptococcusfaecalis) (such as the ATCC-10541 bacterial strain), Staphylococcus cohnis (Staphylococcus cohnii), streptococcus aureus (Staphylococcus aureus) (such as IP 52154 or ATCC-6538 bacterial strain), subtilis (Bacillus subtilis) (ATCC-19659 bacterial strain) (being common hospital bacterial isolates), faecium (Enterococcus facium), Hai Shi faecalis (Enterococcus hirae), thiobacillus ferrooxidant (Thiobacillus ferrooxidans) (such as ATCC 13661 bacterial strains), lactobacillus (Lactobacilli), bacillus acidocldarius (Thermophilic bacilli), Trichophyton between finger (Trychophyton interdigitale) (such as the ATCC-640 bacterial strain), clostridium (Clostridiumsporogenes) (ATCC-3584 bacterial strain), clostridium perfringens (Clostridiumperfringens) (ATCC-13124 bacterial strain), Salmonella typhimurium (Salmonella typhimurium), Listeria monocytogenes (Listeria monocytogenes) etc.Nanometer silver composition of the present invention also has antimycotic activity, comprise for example Candida albicans (Candida albicans) (such as the APCC-2091 bacterial strain), M. smegmatics (Mycobacterium smegmatis) (such as IP 7326 bacterial strains), aspergillus niger (Aspergillus niger) (such as the 218IP bacterial strain), penicillium verruculosum bacterium (Penicilliumverrucosum) etc., also can have Antiparasitic Activity, antagonism is such as Schistosoma haematobium (Schistosomahaematobium) and Schistosoma mansoni (Schistosoma mansoni) etc.
According to a particular implementation of the present invention, described antimicrobial (or antimycotic or parasiticide or antiviral) purposes can be the form of liquid disinfection composition, wherein the nanometer silver composition that produces of aforesaid method can with second antimicrobial agent or the combination of this type of reagent mixture.The suitable example of second antimicrobial agent includes but not limited to: hydrogen peroxide, quaternary ammonium salt, Peracetic Acid and peracid salt (latter the present invention in this article narrates first aspect), and the mixture of any known proportion.Specifically, described combination can provide the synergistic effect of antimicrobial acivity.In an embodiment, described second antimicrobial agent can be the oxidation biocide, such as but not limited to, dioxide peroxide, monochloro amine, hypochlorite, potassium permanganate, iodine or chlorine.According to this embodiment of the present invention, the liquid disinfection composition also can comprise one or more stablizers, such as phosphoric acid, nitric acid, sulfuric acid, Hydrogen bromide or boric acid or its mixture, namely is in the scope that is fit to processing and uses for the pH that adjusts composition.Preferably phosphoric acid especially in the mineral acid stablizer.In the practice, described sour stablizer is incorporated in the commercially available hydrogen peroxide rank with appropriate amount usually.The optional stablizer that uses of the present invention also can be organic carboxyl acid, such as tartrate, citric acid (or its hydrate), phenylformic acid, picolinic acid, niacin and isonicotine acid.Also consider to use the mixture of organic acid and mineral acid for same purpose.When occurring, the amount of described stablizer preferably can effectively be adjusted pH and/or the extended storage stability of liquid disinfection composition.These sterilised liq compositions of the present invention also can comprise at least a being selected from: the composition of tensio-active agent, sanitas or spices (perfume).
The tensio-active agent that is suitable for sanitizing composition of the present invention comprises, such as but not limited to cationic, non-ionic type, facultative or zwitter-ion surface active cpd, preferably be fit to contact food or those compounds of tap water and the mixture of this compounds in related concentrations.The potential use of this paper be multiple nonionogenic tenside.The non-limitative example of anion surfactant comprises, for example, is selected from polyethoxylated and/or poly-propenoxylated glycol, C
8-C
20Fatty acid monoester, anhydrous sorbitol monopalmitate etc.The object lesson of suitable amphoterics comprises 3-sodium dodecyl aminopropionitrile, 3-dodecyl aminopropanesulfonic acid sodium, N-alkyl taurine and trimethyl-glycine.
The sanitizing composition that contains nanometer silver available from the inventive method can be stablized in bio-matrix, can be directly or according to after the further processing mentioned above, and for the treatment of, cleaning or eliminate environmental pollution.For example nano-Ag particles can be scattered in and must remove near the place of bacterium or its by any suitable method or application method.Nanometer silver component in the composition can with the bacterial cell component interaction, also reduce the total cell count of bacterium to acceptable level thereby can effectively destroy them.
Use as the above-mentioned liquid composition that contains nanometer silver can carry out the gas of solid surface or certain volume or liquid cleaning, depollute, sterilize or sterilize.When described liquid composition of the present invention is used for sterilization or sterilization composition (as disperseing to enter liquid or gas), usually under conditions suitable, comprise that concentration and Applicative time use, those skilled in the art are easy to determine these conditions by the standard knowledge of sterilization and sterilization.
When solid surface application the present invention comprises the sterilised liq composition of nanometer silver, for the purpose of safety standard, preferably use the instant diluted formulations, this prescription obtains by following method: concentrate composition and the water of appropriate amount is mixed, then obtain the dilution wetting described solid surface of preparation with institute until solid surface all becomes wet (known to those of skill in the art, depending on the porosity on surface).
Those skilled in the art will know, occur according to solid surface or want the different of the microorganism type that occurs in treatment liq and the gas and amount, comprise the also difference largely of preferred usage quantity of the sterilised liq composition of nanometer silver.
Foregoing uses the liquid composition comprise nanometer silver during as sterilizing agent according to the present invention, the more following application of special recommendation:
-the product that will process immerses in the described composition that comprises nanometer silver,
-sanitizing composition is sprayed to solid surface to be processed, and
-sanitizing composition (dilution or concentrated) is incorporated into (especially swimming-pool water, industrial treatment water, waste water etc.) in the water to be processed.
Therefore, the sterilised liq composition that comprises nanometer silver according to the present invention especially can be used for:
(a) hospital and place, laboratory, industrial site (such as dairy factory, cheese factory, malthouse, distillery, the Workplace of mineral water, wine, alcohol, fruits and vegetables juice, greenhouse, cowshed, hen house and stable); The inside of food, beverage and pharmacy packaging production line, aircraft and ship) and the content in described place, the sterilization of the equipment in described place and instrument and health in the time of especially;
(b) incubator of the sterilization of sterile enclosure such as prematurity animal or germ-free animal growth;
(c) processing of legionella in the air-conditioning system;
(d) sterilization and the health of the pipeline of storage vessel (especially silo) and transmission liquid or solid product such as food (sugar, tea, coffee, cereal, beverage) and animal-feed;
(e) sterilization and the health of swimming pool and other shower set, the at this moment composition of preferred surfactant-free;
(f) production of tap water, transport and the sterilization of stocking system (for example well or storage vessel), at this moment the composition of preferred surfactant-free; And
(g) protect outdoor crop (such as cereal, tomato, banana garden, water culture thing such as Herba Sonchi Arvensis, seed and stem tuber etc.) not to be subjected to bacterium, fungi, virus and parasitic injury.
Except industrial application widely; the optionally high antimicrobial acivity that the inventive method is obtained also has widely domestic use; such as but not limited to water sterilization; remove algae in the water; cleaning product and antimicrobial coatings coating formulation; as be used for medical use or handler with or the nutrition used of animal or other material (special because do not affect or affect minimum for eukaryotic cell or organism); be used for the antimicrobial protection of textile product; be used for preventing from exposing the external application medical preparation that infection or microbial contamination occur tissue; for example; but be not limited to; ointment; salve or lotion, or as chemistry or the catalyzer of other conversion process.By the nanometer silver suspended substance nanometer silver is mixed the above-mentioned various application of realization in polymkeric substance and/or other type coatings.
The 3rd aspect of the present invention relates to probiotic bacterium and other bacterium and prepares the metal nano precipitation; surprisingly; this metal nano precipitation can be used as algicide (anti-chlorella (Chlorella vulgaris); but be not limited to this); be used for tap water, fishpond or pond or swimming-pool water, fresh water or flushing water reservoir, polymkeric substance and paint; topcoating and other material that needs protection prevent soft dirt (aesthetic) or hard scale (material degeneration).The 4th aspect of the present invention relates to probiotic bacterium and other bacterium and prepares the metal nano precipitation, surprisingly, this metal nano precipitation can be used as the dicotyledonous or monocotyledons of weedicide antagonism or resists dissimilar lower plant, such as liver moss, can be diluted in the water or further usefulness machinery, zymetology and/or the processing of physicochemical method.Purpose selection that corresponding plants are carried out is not key parameter of the present invention accordingly.Suitable plant with this purpose comprises dicotyledons such as tobacco (Nicotianatabacum); duck grass (Lamna sp.); soybean (Glycine max); apple; beet; Arabidopis thaliana; clover; petunia; cotton; Radix Dauci Sativae; celery; wild cabbage; cucumber; pepper; (canola) drawn in the Kano; tomato; potato; French beans; flax; cabbage; soybean; lettuce; rape; Cauliflower; spinach; brussels sprouts; jerusalem artichoke; pea; okra; pumpkin; kale; tea tree; coffee and Selaginella tamariscina (Selaginella lepidophylla).Also can comprise monocotyledons, such as paddy rice (rice Oryza sativa), corn, barley, Zea mays, Sunflower Receptacle (Helianthus annuus), wheat, oat, grain, Chinese sorghum, three-coloured amaranth, onion, asparagus and sugarcane.
Above-mentioned aspect of the present invention is particularly useful in following field:
-the growth of inhibition algae in following system: water processing filtering system or the dissimilar sprinkler system of fishpond water, animal and human's tap water system of distribution, gardening water distribution system, pond, swimming pool, pond or swimming pool;
-suppress the surface, comprise the surface that contacts with water, such as the algal grown of hull;
-prevent algae for the treatment of the surface, comprise paint, polymkeric substance and the coating of the algal grown of the higher organism surface that prevents similar plants;
-suppress the growth of liver moss or other undesirable plant, comprise dicotyledonous and monocotyledons, specifically leaf, stem or root system system are exposed to colloidal silver, for example produced by probiotic bacterium according to aforementioned production method and precipitate colloidal silver on it; And
-by coating or alternate manner these surfaces being exposed to colloidal silver suppresses certain plants or weeds growth from the teeth outwards, and described colloidal silver for example comprises, produced by bacterium and precipitates colloidal silver on it according to aforementioned production method.
The following example is in order to illustrating some embodiment of the inventive method and sanitizing composition, but do not impose any restrictions.
The preparation of embodiment 1--nanometer silver
(ATCC 11976 for fermentation lactobacillus Beijerinck 1901AL, LMG 8900, enteron aisle from 8 days large breast feeding babies) (this gets (Oxoid to culture available from the AudioCodes of Britain's Basingstoke at MRS meat soup, Basingstoke, United Kingdom)) lower 37 ℃ of little aerobic condition was cultivated 15 hours in.15 ℃ of centrifugal 10 minutes collecting cells from MRS of 3,000g are used milliQ water washing twice, then are resuspended in to make its final optical density at 600nm (OD in the milliQ water
600) be 1.5.To join in the cell suspension in the 1N sodium hydroxide mother liquor, be respectively 0.05N NaOH and 0.10N NaOH such as final concentration.
Preparation 425mg AgNO in 50mL milliQ water
3With 225mg NH
4The Ag of Cl (I) mother liquor.With this Ag (I) mother liquor of 1 volume join respectively contain 0.05 and the 10 volume cell suspensions of 0.10N NaOH in.25 ℃ these mixtures are hatched mild stirring (per minute 100 turns on the agitator) 30 minutes under visible light.(535mg Ag (0)/L) is deposited on the final solution on the lactobacillus fermentum biological substance, and this paper is called " nanometer silver " or " nanometer-Ag " to obtain 5.0mM Ag (0).With coated lactobacillus fermentum cell centrifugation, by repeated centrifugation, decant and composition is resuspended in fresh milliQ water removes residual and other additive of growth medium three times with the milliQ water washing.Then adjust final nanometer Ag concentration.Then use as required milliQ water diluted composition or with its 3, be resuspended in behind the 000g centrifugal concentrating in the milliQ water.
Embodiment 2-nanometer silver XRD analysis
After gained among the embodiment 1 being had the further 30 ℃ of dryings of biological substance of silver-colored particle, carry out X-ray diffraction (XRD) with the siemens D5000 diffractometer (available from Siemens Company, the Munich, Germany) that is equipped with Prague-Brunt (Bragg-Brentano) optical system.The X-ray is the copper X-x ray tube generation of 1.6kW (40kV, 40mA) by power.Measure between 25-90 degree 2 θ, " tep time ", (tep time) was 1.6 seconds, with the big or small stepping of 0.02 degree.Gained spectrum (not shown) has shown the existence of argent and sodium oxide X-ray diffraction pattern.The latter is that the employed sodium hydroxide of preparation nanometer silver is remaining.
The EDX of embodiment 3-nanometer silver analyzes
After gained among the embodiment 1 being had the further 30 ℃ of dryings of dry biological substance of nanometer silver, carry out energy dispersive X-ray (EDX) analysis with the JSM6100 scanning electronic microscope (available from U.S. joule company (JEOL USA, Inc.)) of the EDX detector that is equipped with corresponding projectile energy 20.0keV resolving power.Analytical results (being expressed as simultaneously quality % and atom %) as shown in table 1, clear showing mainly exists organic substance (because the content of carbon and oxygen is high) and silver, and its combination amounts to and accounts for 91% of dry matter weight.The residue dry substance is comprised of trace elements Ca, Mg, Si, P, S and Cl, mainly is because the mineral substance in the dry bio-matrix is remaining.
Table 1
| Element | % by weight | Atom % |
| C | 55.90±0.28 | 69.18±0.16 |
| O | 26.21±0.08 | 24.35±0.15 |
| Na | 4.97±0.02 | 3.21±0.00 |
| Mg | 0.85±0.06 | 0.52±0.04 |
| Si | 0.19±0.06 | 0.10±0.03 |
| P | 1.64±0.04 | 0.79±0.02 |
| S | 0.22±0.01 | 0.11±0.01 |
| Cl | 0.31±0.03 | 0.13±0.014 |
| Ag | 8.51±0.20 | 1.17±0.03 |
| Ca | 1.22±0.18 | 0.45±0.07 |
The antimicrobial acivity of embodiment 4-nanometer silver on the intestinal bacteria solid growth culture media
Form is that the 100mL Ag concentration of the nanometer silver that is deposited on the lactobacillus fermentum biological substance of acquisition among the embodiment 1 is the silver-colored suspension of 5mM, is plated on to solidify on the growth medium to cultivate intestinal bacteria (get over-Bel's tower Buddhist nun (Luria Bertani) agar on the road).In contrast, the 5mM AgNO of the aseptic milliQ aqueous solution preparation of 100mL 0.1N NaOH
3Be plated on the identical growth medium.This setting is equal to every block of agar plate, and to contain total amount be 0.05mg Ag, or 11mg Ag/m
2Total surface area.Rear concentration with twice repeats this experiment, i.e. every agar plate 0.11mg Ag, or 22mg Ag/m
2Total surface area.
By with these silver-colored suspension bed boards, with uniform Ag (I) NO
3Or the nanometer Ag layer is applied on the curing growth medium.
After growth medium is solidified in pre-treatment like this, with the 2x 10 of 100 μ L physiological solutions (8.5g NaCl/L sterilized water) preparation
6CFU/mL intestinal bacteria suspension is plated on the pretreated agar plate.Then flat board was hatched 24 hours and counted bacterium colony at 30 ℃.Count results as shown in Figure 1.When nanometer Ag concentration is 11mg Ag/m
2With 22mg Ag/m
2The time, on the solid growth culture media of processing, do not detect survival Bacillus coli cells (＜detectability (detection limit) (D.L.)=1x 10
1CFU/ml).Therefore the nanometer Ag of these concentration is processed and is caused Bacillus coli cells from 2x 10
6CFU/ml is reduced by at least in 1x 10
1CFU/ml (D.L.).As Ag (I) NO
3Concentration is 11mg Ag/m
2With 22mg Ag/m
2The time, Bacillus coli cells is respectively from 2x 10
6CFU/ml obviously is reduced to 4x 10
2CFU/ml and 1x 10
2CFU/ml.
In contrast, concentration is 2x 10
6The intestinal bacteria suspension of CFU/mL is plated on and is untreated, namely do not contain on the growth medium of Ag, and be plated on 100 μ L and only contain on the same growth medium of aseptic mQ water treatment of lactobacillus fermentum ATCC 11976, it is identical that its concentration and nanometer silver are processed, but do not contain nanometer Ag.Do not observe the restraining effect to the total count of these bacteriums.Therefore, the nanometer Ag of observing and Ag (I) restraining effect is attributable to Ag and processes, and is not employed lactobacterium strain in treatment step or the experiment.
In the embodiment 5 – suspensions to the antimicrobial acivity of various pathogenic bacteria
Detection contains the survival of the pathogenic colon bacillus (Escherichia coli), Salmonella typhimurtum (Salmonella typhimurium), streptococcus aureus (Staphylococcus aureus) and Listeria monocytogenes (Listeria monocytogenes) culture that dilute in the physiological solution of the nanometer Ag composition that obtains among different concns (0mg/L, 0.10mg/L, 1.0mg/L, 10mg/L and the 50mg/L) embodiment 1.Nanometer Ag is applied to contain in the physiological solution of culture alive of a kind of above-mentioned malignant bacteria.The physiological solution that dissolving 8.5g NaCl obtains in 1 premium on currency and bacterial cell etc. ooze, therefore can be because of the osmotic pressure cell killing.Control treatment comprises the bacterial cultures by the physiological solution preparation that does not contain nanometer Ag.
The mother liquor of 100mg nanometer Ag/L in the preparation mQ water, and the bacterial cultures that joins the physiological solution dilution makes its final nanometer Ag concentration as shown in table 2.
Each above-mentioned pathogenic species all carries out independently re-treatment, the liquid meat soup that contains exponential phase growth bacterium of " bacterial cultures " expression dilution, and being diluted to final concentration of cells with physiological solution is 10
4-10
5CFU/ml.Each is processed to carry out in duplicate.All hatching all carried out in aseptic test tube with cover, and concussion was cultivated 72 hours under 37 ° of C.After hatching, from each test tube, get 100 μ L and be plated on trypticase soy agar (TSA) solid growth culture media and count bacterium colony.Count results for difference test pathogenic agent is as shown in table 2.
Table 2
It is that the nanometer Ag of 1mg/L was enough to the cell concn of intestinal bacteria, streptococcus aureus and Salmonella typhimurtum is reduced to＜10CFU/mL (namely being lower than detectability) in 72 hours that table 2 shows available from embodiment 1 concentration.Be that 0.10mg/L has observed significant necrocytosis in concentration.As for the listeria bacteria, the 10mg/L nanometer Ag is down under the detectability viable cell concentrations.Therefore we reach a conclusion: the nanometer Ag that obtains from embodiment 1 is 1.0mg/L or when lower in the concentration of liquid cell suspension, can significantly effectively reduce survival pathogenic bacteria in the liquid as potent biocide.
Nanometer Ag and San Francisco Bay halogen worm (Artemia franciscana) coupling in the embodiment 6-suspension
Antimicrobial acivity
Prepare aseptic artificial seawater (Instant Ocean by autoclaving with milliQ water
R, available from U.S.'s aquarium system (Aquarium Systems USA)).All are processed in dividing the 50mL Falcon test tube that the aseptic artificial seawater of 20mL is housed and carry out.20 artemia naupliis that each processing (operating in triplicate) contains the preparation of 20mL artificial seawater wherein add 10
5CFU/mL (colony-forming unit) Vibrio campbellii (Vibrio campbellii) LMG21363 and/or available from the nanometer Ag of embodiment 1, final concentration is as shown in table 3.The pathogenic bacterium Vibrio campbellii is hatched with its host's organism San Francisco Bay halogen worm.
Be set as follows test:
-San Francisco Bay halogen worm+10
5The CFU/ml Vibrio campbellii
-San Francisco Bay halogen worm+10
5CFU/ml Vibrio campbellii+100mg nanometer Ag/L
-San Francisco Bay halogen worm+10
5CFU/ml Vibrio campbellii+10mg nanometer Ag/L
-San Francisco Bay halogen worm+10
5CFU/ml Vibrio campbellii+1.0mg nanometer Ag/L
-San Francisco Bay halogen worm+10
5CFU/ml Vibrio campbellii+0.1mg nanometer Ag/L
-San Francisco Bay halogen worm+0.10mg nanometer Ag/L
-San Francisco Bay halogen worm+100mg nanometer Ag/L
-San Francisco Bay halogen worm+10mg nanometer Ag/L
-San Francisco Bay halogen worm+1.0mg nanometer Ag/L, and
-San Francisco Bay halogen worm+0.10mg nanometer Ag/L
After hatching 48 hours, contain the concentration of Vibrio campbellii in the sterilization artificial seawater of San Francisco Bay halogen worm by being plated on specific vibrios grown cultures based assays.Below table 3 has shown average treatment result (wherein D.L. refers to detectability).
Table 3
Also noteworthy is that with untreated control and compare, concentration be 0.10 and the 1.0mg/L nanometer Ag survival rate of San Francisco Bay halogen worm is had no significant effect (80%).This shows that the nanometer Ag that produces according to embodiment 1 does not have toxicity or restraining effect to high organism under these concentration.
Embodiment 7 – measure effective duration of contact of antimicrobial acivity
The purpose of this test is to measure nanometer silver composition and the suitable duration of contact that is diluted in pathogenic bacteria culture intestinal bacteria (Escherichia coli), Salmonella typhimurtum (Salmonellatyphimurium), streptococcus aureus (Staphylococcus aureus) and Listeria monocytogenes (Listeria monocytogenes) in the physiological solution among the embodiment 1, take acquisition concentration as 0.1 and effective antimicrobial acivity during 1mg/LAg.
These nanometer Ag concentration are applied in the bacterial cultures of physiological solution (dissolving 8.5gNaCl in 1 premium on currency) preparation, physiological solution and bacterial cell etc. oozes, therefore can be because of the osmotic pressure cell killing.Control treatment is included in the bacterial cultures in the physiological solution that does not contain the nanometer Ag composition.
The 100mg nanometer Ag of preparation mQ water preparation/L mother liquor also adds bacterial cultures (with equivalent in meaning among the embodiment 5) in the physiological solution of appropriate amount so that required final concentration nanometer Ag to be provided.
All hatch (in duplicate) all carry out in aseptic test tube with cover, and 37 ℃ of concussions are cultivated, and carry out cell counting (sampling accident) in different duration of contact.When each sampling accident, from each processes sample, get 100 μ L and be plated on trypticase soy agar (TSA) solid growth culture media and count bacterium colony.Result after 15 hours, 16 hours, 17 hours, 18 hours and 40 hours is shown in lower tabulation 4 (wherein ND represents not detect, and namely is lower than detectability).
Table 4
The nanometer silver composition of the different silver of embodiment 8-preparation and biological substance dry cell weight weight ratio
With the liquefied ammonia (NH of 28% volume in the water
3) preparation silver (I) mother liquor, final concentration is 425g AgNO
3/ L (=50mM AgNO
3).Prepare as described in Example 1 the lactobacillus fermentum culture.
2.8g (weight in wet base) eccentric cell agglomerate is resuspended in 3 kinds of differences to be measured in the milliQ water of (50ml, 100ml and 1L) to obtain to be called the reaction mixture of A, B and C.
The NaOH mother liquor of 1N MilliQ water preparation is added in each test tube, in above-mentioned suspension, to obtain normality 0.10N NaOH.
Then, following adding silver (I) mother liquor:
-reaction mixture A: add 0.24mL silver (I) mother liquor and obtain final concentration 1.30g Ag/L (or 12mM) Ag.Precipitin reaction (reddish-brown precipitation) almost occurs on the 56g/L biological substance (weight in wet base) immediately.Suppose that the average dry weight of centrifugal biological substance compares between 10-30%, thereby acquisition is silver-colored and dry cell weight compares between 1:4 and 1:12.
-reaction mixture B: add 2.4mL silver (I) mother liquor and obtain final concentration 5.78g Ag/L (or 55mM) Ag.Precipitin reaction (reddish-brown precipitation) almost occurs on the 28g/L biological substance (weight in wet base) immediately.Because the dry weight of centrifugal biological substance on average accounts for 10-30%, thereby the ratio of acquisition silver and dry cell weight is between 2:1 and 0.7:1.PH is 11.6 in this reaction.
-reaction mixture C: add 24mL silver (I) mother liquor and obtain final concentration 5.78g Ag/L (or 55mM) Ag.2.8g/L almost precipitate immediately on the biological substance (weight in wet base).Because the dry weight of centrifugal biological substance on average accounts for 10-30%, thereby the ratio of acquisition silver and dry cell weight is between 20:1 and 7:1.
With reaction mixture sat 30 minutes, then collect the nanometer Ag composition that forms.
Gained nanometer Ag precipitation is with 15 ℃ 3 of biological substance, centrifugal 10 minutes of 000g, and then flushing is removed any residual ammonia and water soluble ingredient in the production process twice in milliQ water.Then analyze nanometer Ag purifying agglomerate product (embodiment 9), or further be diluted to the nanometer Ag concentration that is fit to further test with milliQ water.
Embodiment 9 – produce nanometer Ag when the ratio of silver and biological substance dry cell weight is 0.7:1
XRD analysis
According to embodiment 8 silver with the biological substance dry cell weight
ItThan for preparation under the condition of 0.7:1 contains the biological substance of nano-Ag particles, in 100 ℃ of baking ovens, after dry 24 hours, carry out as described in Example 2 XRD analysis.In this XRD spectrum, only detect the X-ray diffraction pattern of silver metal.Because estimate that safely quality in the desciccate is less than 5% crystal trace elements and can't detects in XRD, be in Ag (0) state so approximately estimate the silver at least 95% that detects in the XRD analysis.
Embodiment 10 – H
2
O
2
Nanometer silver is carried out post-processed
With containing 30% (volume) H
2O
2The aqueous solution nanometer Ag agglomerate according to embodiment 1 or embodiment 8 resulting washings is carried out post-processed.Agglomerate is suspended in H for this reason
2O
2Obtain the concentration height to 6gAg/L H
2O
2(30%).Obtain more stable precipitation.Then the suspension that further dilutes gained precipitation with milliQ water obtains the nanometer Ag for the suitable concn of follow-up test.
Prepare respectively nanometer Ag preparation (this paper is called respectively sample A, B and C) according to embodiment 8 is described with different silver and biological substance dry cell weight ratio 7:1,1:10 and 0.7:1.In addition, further process according to embodiment 10 described methods with H2O2 with the nanometer Ag prepared product that silver and biological substance dry cell weight ratio 0.7:1 obtain, thereby obtain the 4th sample D.
In order to evaluate silver and biological substance dry cell weight ratio to the impact of nanometer silver product antimicrobial acivity, in sterile physiological solution, prepare 1x 10
4CFU/mL salmonella typhimurium cell suspension, and be sub-packed in the different test tubes.Sample A, B, C and D are added test tube until obtain the nanometer Ag that final concentration is 0.05mg/L (or 50ppb) in each test tube.In contrast, with bacterial cultures and 0.5ppm AgNO
3Hatch or do not hatch with any silver.Cover test tube and at 37 ℃ of duplicate oscillation incubations.After hatching 4.5 hours, take out sample and also in physiological solution, carry out serial dilution, be plated on the TSA substratum after, 37 ℃ of overnight incubation are measured to carry out total Salmonellas counting.Fig. 3 has shown the result of these countings, represents with average cell counting and twice independent standard deviation that repeats.Under embodiment 8 described method gained 0.05ppm nanometer silver existence conditions, hatch and observe the minimizing of obvious bacterial cell counting after 4.5 hours.Fig. 3 show silver and the dry cell weight ratio higher, the antimicrobial acivity of gained nanometer Ag is more weak.Equally, utilize H
2O
2Process the nanometer Ag product and significantly increase its antimicrobial acivity.
Embodiment 12 – transmission electron microscope methods detect the nanometer silver granularity
In order to prepare the section for tem analysis, with 0.1M cacodylate damping fluid (pH 7.4) the fixation of bacteria agglomerate that contains 2.5% glutaraldehyde and 2% formaldehyde, and be embedded in 3% low melting-point agarose (from Fick laboratory (Difco Laboratories), the Michigan, USA Detroit).Fixing after in 1% perosmic anhydride, sample being carried out.Between fixing step or afterwards, use the distilled water wash sample.Then, in the ethanol that increases concentration and anhydrous propylene oxide, sample is dewatered.After in epoxy resin medium (Epon-Spurr medium), carrying out embedding, sample blocks is repaiied at TM60 and is repaiied piece (from Ri Chite-river company (Reichert-Jung A.G), Austria Vienna) in the module unit to obtain cut surface as 0.5X 1mm
2-1X 2mm
2Section, and with the Ultracut cutting machine (from Ri Chite-river company (Reichert-Jung A.G), Austria Vienna) of cutting into slices gold is cut to the ultrathin section(ing) in the silver-colored black interface color gamut.Section is positioned on the coated copper mesh of polyvinyl acetal and carbon (200 order).In case finish, with 2% uranyl acetate thin section is dyeed, and detect the ultrastructure of cell with lead citrate.Chemical and net purchase are from Ah loud, high-pitched sound's scientific company (Agar Scientific) (Britain Tan Site).Utilize the EM208S transmission electron microscope to carry out imaging (from FEI Co. of Eindhoven, Holland) at 80kV.
Obtain the TEM image (not shown) of the nano-Ag particles that embodiment 8 described reaction mixture A, B and C obtain.These image confirmings obtain the spherical nano-silver particle at composition, and its existence form is the precipitation on bacterial cell surface or the suspended substance between biological substance.
The nanometer silver granularity that-reaction mixture A (ratio 1:10) produces: for 35 nano particles measuring, diameter range 3.3nm-72nm, mean value are 14nm, granule surface area scope 6.4-2,996nm
2, so sphericity scope 0.14-0.97;
The nanometer silver granularity that-reaction mixture B (ratio 1:1) produces: for 202 nano particles measuring, diameter range 3nm-116nm, mean value are 15nm, granule surface area scope 6-4,805nm
2, so sphericity scope 0.12-0.96;
The nanometer silver granularity that-reaction mixture C (ratio 1:10) produces: for 56 nano particles measuring, diameter range 3.3nm-56nm, mean value are 16nm, granule surface area scope 6.4-1,841nm
2, so sphericity scope 0.15-0.95.
Embodiment 13 – prepare the colloidal nano gold
With milliQ water preparation gold (III) mother liquor, final concentration concentration is 7.5g AuCl
3/ L.Obtain the lactobacillus fermentum culture according to embodiment 1.
The eccentric cell agglomerate of weight in wet base 2.5g is joined in the 100mL milliQ water.
The sodium hydroxide mother liquor of 1N milliQ water preparation added to obtain conventional final concentration in the above-mentioned suspension be 0.10N NaOH.
Then adding 10mL gold (III) mother liquor acquisition final concentration in this suspension is 75mg Au (III)/100mL, and existence form is AuCl3-Au (3.8mM Au).Au in 4 hours (0) finishes and is deposited on the 2.5g/100mL biological substance (weight in wet base).Since centrifugal biological substance dry weight on average accounts for 10-30%, therefore obtain gold and dry cell weight ratio between 1:3 and 1:10.
Allow reaction to continue to carry out 4 hours, then collect nm gold particles.Remove from the water soluble ingredient in the production process for 2 times with centrifugal 10 minutes of 15 ℃ of this purple precipitation 3,000g and with the flushing of milliQ water.
The XRD analysis of embodiment 14 – nanometer gold
The biological substance that contains gold grain that contains embodiment 13 after dry 24 hours, carries out XRD analysis with the siemens D5000 diffractometer that is equipped with Prague-Brunt (Bragg-Brentano) optical system as described in Example 2 in 100 ℃ of baking ovens.The gained spectrogram shows only to have Au as shown in Figure 4
0The X-ray diffraction peak.
Biogenic sediment efficient and biological thing under the different silver of embodiment 15 – and the biological substance dry cell weight ratio
The recovery of matter
Be the impact of Ag (0) nano particle in order to assess biological substance on Ag (I) biological reducing, detect under the different Ag:CDW ratios behind the biological reducing on biological substance and the recovery in the solution.
Prepare as described in Example 8 different Ag:CDW ratio nanometer silver preparations.Hatched 4 hours at biological substance and Ag (I), 7,000g measures silver-colored recovery percent after separating solvable phase (in the solution) and precipitated phase (on the biological substance) in centrifugal 10 minutes.This result of study is as shown in table 8 below.
Table 8
| Sample (Ag:CDW ratio) | Mg Ag/L in the solution | Mg Ag/L on the biological substance | The total recovery |
| A(1:10) | 76 | 1,295 | 100% |
| B(1:1) | 819 | 5,308 | 100% |
| C：Ag:CDW=10:1 | 1,612 | 4,844 | 100% |
These results clearly show that when the Ag:CDW ratio was low, the silver recovery of the relevant particle form of biological substance was higher.For example the Ag:CDW ratio is that 1:10 obtains reclaiming acceptable Ag from Ag (I) solution that the biological reducing method is processed
0(about 95%).
Embodiment 16 – are without the algae removal characteristic of the nanometer silver preparation of hydrogen peroxide aftertreatment
Prepare nanometer silver preparation with silver with biological substance dry cell weight ratio 1:4 according to embodiment 8 described methods.
In order to assess the algae removal effect of said preparation, containing 10mL BG11 substratum (such as Stanier etc., Bacteriol.Rev. (1971) 35:171-205 is described) test tube in inoculation enter 0.5mL liquid B G11 Fast Growth chlorella culture, cultivate at 20 ℃, 65% relative humidity and 1000Lux (16 hours/day).Utilize spectrophotometry evaluation growth after 2 weeks.By the dosage in the test tube, detect the nanometer silver preparation of different concns, scope is from 20mg Ag/L-0.01mg Ag/L.Minimum test concentrations when observing complete organism growth and suppressed fully is the MIC value.The MIC pH-value determination pH of the anti-chlorella growth of this nanometer silver preparation is 0.125mg Ag/L.
Claims (8)
1. method of producing colloidal metal or colloidal metal compound at bacterial film with bacterium, described method is by under controlled pH described bacterium being contacted with described metal or metallic compound.
2. the method for claim 1, the metal in described metal or the metallic compound are silver.
3. the method for claim 1, the metal in described metal or the metallic compound is selected from: gold, zinc, mercury, copper, palladium, platinum and bismuth.
4. such as each described method in the claims 1 to 3, described bacterium belongs to Bacterium lacticum.
5. such as each described method in the claim 1 to 4, described controlled pH is 8 to 12.
6. such as each described method in the claim 1 to 5, wherein the time of contact is 1 second to 30 minutes.
7. method as claimed in claim 2, described metallic compound is Silver Nitrate.
8. method as claimed in claim 3, described metallic compound gold trichloride.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US81857906P | 2006-07-05 | 2006-07-05 | |
| US60/818,579 | 2006-07-05 | ||
| GBGB0623925.5A GB0623925D0 (en) | 2006-11-30 | 2006-11-30 | Method for producing silver nanoparticles |
| GB0623925.5 | 2006-11-30 |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007800251987A Division CN101506371B (en) | 2006-07-05 | 2007-07-05 | Method for producing metal nanoparticles |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102978241A true CN102978241A (en) | 2013-03-20 |
Family
ID=37671599
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012104486448A Pending CN102978241A (en) | 2006-07-05 | 2007-07-05 | Method for producing metal nanoparticles |
Country Status (6)
| Country | Link |
|---|---|
| CN (1) | CN102978241A (en) |
| AT (1) | ATE537266T1 (en) |
| BR (1) | BRPI0714227A2 (en) |
| ES (1) | ES2379017T3 (en) |
| GB (1) | GB0623925D0 (en) |
| RU (1) | RU2460797C2 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102962467A (en) * | 2012-10-26 | 2013-03-13 | 上海交通大学 | Method for preparing noble metal nano material with adjustable particle size by bacteria |
| CN104694398A (en) * | 2015-03-04 | 2015-06-10 | 大连理工大学 | Trichosporon montevideense and application of trichosporon montevideense in synthesis of gold nanoparticles |
| CN105780067A (en) * | 2016-02-01 | 2016-07-20 | 中国科学院生态环境研究中心 | Method for in-situ synthesis of three-dimensional nanometer palladium catalyst layer through electrode activity biological membrane and application |
| CN107641632A (en) * | 2017-10-18 | 2018-01-30 | 福州大学 | A kind of method with the carbon-based point of Microbe synthesis |
| US10363218B1 (en) | 2019-01-03 | 2019-07-30 | King Saud University | Synthesis of probiotic nanoparticles |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014200380A1 (en) * | 2013-06-10 | 2014-12-18 | Общество с ограниченной ответственностью "Нанобиотех" | Antiseptic preparation and methods for producing and using same |
| US20170247720A1 (en) * | 2014-11-10 | 2017-08-31 | Cristhian Rene LOPEZ NAVARRO | Biological process for producing magnetic nanoparticles |
| RU2614118C1 (en) * | 2015-11-09 | 2017-03-22 | Федеральное государственное бюджетное учреждение науки Институт химической физики им. Н.Н. Семёнова Российской академии наук (ИХФ РАН) | Application of anabaena sp. pcc 7120 strain for silver nanoparticles production |
| RU2711559C1 (en) * | 2019-04-29 | 2020-01-17 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Российский химико-технологический университет имени Д.И. Менделеева" (РХТУ им. Д.И. Менделеева) | Method of producing colloidal solution of silver nanoparticles with plant leaves extracts |
| RU2734350C1 (en) * | 2020-01-16 | 2020-10-15 | Федеральное государственное бюджетное учреждение "48 Центральный научно-исследовательский институт" Министерства обороны Российской Федерации | Disinfectant |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050013759A1 (en) * | 1996-08-14 | 2005-01-20 | Grow Ann E. | Practical method for preparing inorganic nanophase materials |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2278669C1 (en) * | 2004-11-09 | 2006-06-27 | Иркутский институт химии им. А.Е. Фаворского Сибирского отделения Российской академии наук (ИрИХ СО РАН) | Agent possessing antibacterial activity |
-
2006
- 2006-11-30 GB GBGB0623925.5A patent/GB0623925D0/en not_active Ceased
-
2007
- 2007-07-05 RU RU2009103776/10A patent/RU2460797C2/en not_active IP Right Cessation
- 2007-07-05 CN CN2012104486448A patent/CN102978241A/en active Pending
- 2007-07-05 BR BRPI0714227-7A patent/BRPI0714227A2/en not_active IP Right Cessation
- 2007-07-05 AT AT07765165T patent/ATE537266T1/en active
- 2007-07-05 ES ES07765165T patent/ES2379017T3/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050013759A1 (en) * | 1996-08-14 | 2005-01-20 | Grow Ann E. | Practical method for preparing inorganic nanophase materials |
Non-Patent Citations (3)
| Title |
|---|
| BINOJ NAIR AND T. PRADEEP: "Coalescence of Nanoclusters and Formation of Submicron Crystallites Assisted by Lactobacillus Strains", 《CRYSTAL GROWTH & DESIGN》 * |
| DEENDAYAL MANDAL等: "The use of microorganisms for the formation of metal nanoparticles and their application", 《APPL MICROBIOL BIOTECHNOL》 * |
| TANJA KLAUS等: "Silver-based crystalline nanoparticles,microbially fabricated", 《PNAS》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102962467A (en) * | 2012-10-26 | 2013-03-13 | 上海交通大学 | Method for preparing noble metal nano material with adjustable particle size by bacteria |
| CN102962467B (en) * | 2012-10-26 | 2015-04-01 | 上海交通大学 | Method for preparing noble metal nano material with adjustable particle size by bacteria |
| CN104694398A (en) * | 2015-03-04 | 2015-06-10 | 大连理工大学 | Trichosporon montevideense and application of trichosporon montevideense in synthesis of gold nanoparticles |
| CN104694398B (en) * | 2015-03-04 | 2017-05-03 | 大连理工大学 | Trichosporon montevideense and application of trichosporon montevideense in synthesis of gold nanoparticles |
| CN105780067A (en) * | 2016-02-01 | 2016-07-20 | 中国科学院生态环境研究中心 | Method for in-situ synthesis of three-dimensional nanometer palladium catalyst layer through electrode activity biological membrane and application |
| CN107641632A (en) * | 2017-10-18 | 2018-01-30 | 福州大学 | A kind of method with the carbon-based point of Microbe synthesis |
| US10363218B1 (en) | 2019-01-03 | 2019-07-30 | King Saud University | Synthesis of probiotic nanoparticles |
Also Published As
| Publication number | Publication date |
|---|---|
| RU2009103776A (en) | 2010-08-10 |
| BRPI0714227A2 (en) | 2013-01-29 |
| GB0623925D0 (en) | 2007-01-10 |
| ATE537266T1 (en) | 2011-12-15 |
| RU2460797C2 (en) | 2012-09-10 |
| ES2379017T3 (en) | 2012-04-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101506371B (en) | Method for producing metal nanoparticles | |
| CN102978241A (en) | Method for producing metal nanoparticles | |
| Chaudhary et al. | Metal/metal oxide nanoparticles: toxicity, applications, and future prospects | |
| Ahmed et al. | Green synthesis and characterization of zirconium oxide nanoparticles by using a native Enterobacter sp. and its antifungal activity against bayberry twig blight disease pathogen Pestalotiopsis versicolor | |
| Ogar et al. | Antifungal properties of silver nanoparticles against indoor mould growth | |
| US9162219B2 (en) | Fenton reaction catalyst produced using reducing organic substance as raw material | |
| KR102124574B1 (en) | Method for producing natural complex composition having prevention of pests, offensive odor removal of barn, and sterilization effect | |
| Khaydarov et al. | Silver nanoparticles: Environmental and human health impacts | |
| MX2011011862A (en) | Nanostructural composition of biocide. | |
| Safavi et al. | The study of nano silver (NS) antimicrobial activity and evaluation of using NS in tissue culture media | |
| Doolotkeldieva et al. | Biological Activity of Ag and Cu Monometallic Nanoparticles and Ag‐Cu Bimetallic Nanocomposites against Plant Pathogens and Seeds | |
| Granke et al. | Effects of temperature, concentration, age, and algaecides on Phytophthora capsici zoospore infectivity | |
| Nematollahi et al. | Diazinon reduction in food products: A comprehensive review of conventional and emerging processing methods | |
| Ghasemi et al. | Biosynthesis of silver nanoparticles using Pseudomonas canadensis, and its antivirulence effects against Pseudomonas tolaasii, mushroom brown blotch agent | |
| Abd El-Lateef et al. | Facile green fabrication of copper/polyvinylidene fluoride nanocomposite for pathogens disinfection and dye degradation in wastewater | |
| Tanwar et al. | Role and effects of nanotechnology used in pesticides and agriculture field | |
| Mu | Stability and Toxicity of Cerium Oxide Nanoparticles in Water-Soil-Plant Systems | |
| Husain et al. | Emerging role of biosynthesized silver nanoparticles for agricultural applications | |
| Nsengumuremyi et al. | The Potential Application of Nanoparticles on Grains during Storage: Part 2–An Overview of Inhibition against Fungi and Mycotoxin Biosynthesis | |
| Doolotkeldieva et al. | Research Article Biological Activity of Ag and Cu Monometallic Nanoparticles and Ag-Cu Bimetallic Nanocomposites against Plant Pathogens and Seeds | |
| Madhan et al. | Eco-friendly Nano Colloids for Enhanced Black gram (Vigna mungo) Seed Viability: Experimental and Computational Analysis | |
| Xin | Development, Characterization and Evaluation of Polymeric Nanoparticles for Application in Agriculture | |
| Raza et al. | Exploring the potential of metallic and metal oxide nanoparticles for reinforced disease management in agricultural systems: A comprehensive review | |
| Oktarina | The effect of silver nanoparticles on Trichoderma harzianum, Rhizoctonia spp., and fungal soil communities | |
| Deepa et al. | Activity Of Bi2o3 Nps Helps For The Drug Development On Chenopodium Album |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20130320 |