CN103250739A - Preparation method of oxidized grapheme/silver particle nanometer compound and application thereof - Google Patents
Preparation method of oxidized grapheme/silver particle nanometer compound and application thereof Download PDFInfo
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- CN103250739A CN103250739A CN2012102954161A CN201210295416A CN103250739A CN 103250739 A CN103250739 A CN 103250739A CN 2012102954161 A CN2012102954161 A CN 2012102954161A CN 201210295416 A CN201210295416 A CN 201210295416A CN 103250739 A CN103250739 A CN 103250739A
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Abstract
The invention provides a preparation method of oxidized grapheme/silver particle nanometer compound and application thereof, and an oxidized grapheme solution is prepared by an improved Hummers method, and silver nitrate solid powder is added and boiled, and trisodium citrate is added as reducing agent and stabilizing agent, and the solution is boiled for 1 hour continuously and washed out for 3 times by a ultrafiltration pipe. Compared with the prior art, the preparation method provided by the invention is simple and fast, and the prepared nanometer compound has the advantages of stable structure, good water-solubility and good antibiosis activity, and the product can be used as antibiosis material with high efficiency and safety.
Description
Technical field
The present invention relates to pharmaceutical preparation, be specifically related to preparation method and the application of graphene oxide/silver-colored particle nano-complex.
Background technology
Microbe species is various, comprise prokaryotic micro-organisms (as bacterium), eukaryotic microorganisms (as fungi, algae and protozoon) and acellular biology (as virus) three major types, their individualities are small, but closely bound up with human lives, production, existence, relate generally to numerous areas such as health, food, medicine, industrial or agricultural, environmental protection.Wherein, the pathogenic microbes especially infection of bacterium has brought great harm for aquaculture and human health, has caused enormous economic loss.Therefore, research and the practical application about antibiosis always is subjected to extensive concern.Traditional antibacterial agent comprises antibiotic and chemosterilant (as free chlorine, chloramines, ozone etc.).Antibiotic since coming out just with its efficiently fast bactericidal effect obtain widely applying in the medical and health industry, bringing into play important role aspect the human health protecting.Yet because antibiotic abuse, the resistance problem becomes and is on the rise and gives prominence to, as discovers that staphylococcus aureus to the benzyl penicillin resistance is up to more than 85%.Chemosterilant can be realized producing in enormous quantities, but its toxicity problem has limited its application in clinical.Based on above problem, work out a kind of new type of safe efficiently anti-biotic material have extremely important practical significance and using value.
In recent years, the develop rapidly of nanoscale science and technology has all produced profound influence to every field, and nano material relies on its unique character to demonstrate great application prospect at numerous areas as industry, biomedical sector etc.Discover that some nano materials have good antibacterial activity and are difficult for the generation drug resistance, as gold nano grain, silver nano-grain, Zinc oxide nanoparticle, copper and oxide nano particles thereof etc.Wherein, silver just is acknowledged as since ancient times has anti-microbial property, silver nano-grain then demonstrates more desirable antibacterial activity, has good biological safety simultaneously, so silver nano-grain has broad application prospects in fields such as packaging for foodstuff, purification of water quality and health systems.In addition, discover with nano materials such as list/multi-walled carbon nano-tubes, active carbon, Graphenes as substrate and holder other nano particles of growing, not only can solve dissolubility and the stability problem of nano particle, can also make both functions act synergistically, the nano-complex that obtains is like this used by big quantity research in many aspects.
At present, with respect to other nano anti-biotic materials, graphene oxide/silver-colored particle nano-complex (GO-Ag) is owing to the antibacterial activity of its special nature and significantly enhancing is widely studied.Graphene oxide/the preparation principle of silver-colored particle nano-complex is that silver ion reduction is become silver nano-grain, thereby be grown in the graphene oxide substrate, concrete grammar comprises a stage reduction method, benzene-water two-step reaction method and silver mirror reaction method etc., related reductant comprises formaldehyde, potassium hydroxide, PVP, hydroquinones, boron hydracid sodium etc.The reductant that uses mostly in view of existing preparation method is chemical organic reagent, and toxicity is bigger, harm humans health and ecotope, and the present invention aims to provide the novel preparation method of a kind of graphene oxide/silver-colored particle nano-complex.
Summary of the invention
For overcoming deficiency of the prior art, the invention provides preparation method and the application of graphene oxide/silver-colored particle nano-complex, be a kind of anti-biotic material of highly effective and safe.
For realizing above-mentioned technical purpose, reach above-mentioned technique effect, the present invention is achieved through the following technical solutions:
The preparation method of graphene oxide/silver-colored particle nano-complex may further comprise the steps:
Step 1) prepares the graphite oxide aqueous solution with improved Hummers method:
The solid sodium chloride of 1g expanded graphite and about 30 times of weight is ground to no obvious visible particle shape thing, adds the water suction filtration, flush away sodium chloride is with the graphite dry for standby of porphyrize;
The about 30mL concentration of adding is 98.3% the concentrated sulfuric acid in flask, put into the porphyrize graphite of having dried, stirred 8 hours under the room temperature, then mixture is placed ice bath, add about 3g potassium permanganate, slowly be warming up to 115 ℃, slowly add about 150mL water, add about 10mL30% hydrogen peroxide and stirring afterwards, the centrifugal upper water solution of removing is washed to solution repeatedly and no longer occurs till the precipitation;
Be the graphene oxide hydrosol that the ultrasonic 30min of graphene oxide suspension of 3mg/mL obtains clarifying with 5mL concentration, press then that final concentration 0.12 g/mL adds the NaOH solid and 50 ℃ of stirred in water bath 4 hours, add concentrated hydrochloric acid afterwards and adjust the pH value of solution value near 1, repeated hydrogenation oxygen radical ion not, wash repeatedly and centrifugal purification with distilled water, obtain the water-soluble good not graphite oxide aqueous solution of alkalization;
Step 2) in described graphene oxide solution, adds the silver nitrate pressed powder, boil;
Step 3) adds trisodium citrate as reductant and stabilizing agent, then again 100 ℃ boil 1h, ultrafiltration pipe wash-out three times is removed unconjugated silver-colored particle and other intermediate products, can obtain graphene oxide/silver-colored particle nano-complex.
Further, described graphene oxide/silver-colored particle nano-complex is in the application of antibiosis.
Compared with prior art, the present invention has following beneficial effect:
1, method reaction of the present invention is simple, mild condition, and preparation is fast;
2, employed reductant belongs to environmental type among the present invention, and toxicity is low, and is pollution-free;
3, the graphene oxide of the present invention's generation/silver-colored particle nano-complex stability is high, good water solubility, and high safety, and need not follow-up surface-functionalized modification, namely produce and namely use;
4, graphene oxide/silver-colored particle composites all has significant antibacterial activity to Gram-negative bacteria Escherichia coli and gram-positive bacteria staphylococcus aureus, and colibacillary antibacterial action is better than staphylococcus aureus.
Description of drawings
Fig. 1 is the transmission electron microscope picture (TEM) of 0.65 GO-Ag nano-complex for silver-colored particle and graphene oxide molar ratio;
Fig. 2 is the transmission electron microscope picture (TEM) of 1 GO-Ag nano-complex for silver-colored particle and graphene oxide molar ratio;
Fig. 3 is the transmission electron microscope picture (TEM) of 2 GO-Ag nano-complex for silver-colored particle and graphene oxide molar ratio;
Fig. 4 is the influence of the Escherichia coli growth activity of the GO-Ag of three kinds of different ratios;
Fig. 5 is the influence of the staphylococcus aureus growth activity of the GO-Ag of three kinds of different ratios;
Fig. 6 is the variation of Escherichia coli clump count under the variable concentrations GO-Ag;
Fig. 7 is the variation of staphylococcus aureus clump count under the GO-Ag of variable concentrations;
Fig. 8 is 2.5,5 for the GO-Ag nano-complex at final concentration, during 10ug/ml to the influence of Hela cells growth activity.
Embodiment
Below with reference to the accompanying drawings and in conjunction with the embodiments, describe the present invention in detail.
Preparation anti-biotic material graphene oxide/silver-colored particle nano-complex (GO-Ag).
Do the most initial raw material with the 1g expanded graphite, the solid sodium chloride that adds about 30 times of weight is ground to no obvious visible particle shape thing, adds the water suction filtration, and flush away sodium chloride is with the graphite dry for standby of porphyrize.In flask, add about 30mL concentrated sulfuric acid (98.3%), put into the porphyrize graphite of having dried, stirred 8 hours under the room temperature, then mixture is placed ice bath, add about 3g potassium permanganate, slowly be warming up to 115 ℃, slowly add about 150mL water, add about 10mL30% hydrogen peroxide and stirring afterwards, the centrifugal upper water solution of removing is washed to solution repeatedly and no longer occurs till the precipitation.Be the graphene oxide hydrosol that the ultrasonic 30min of graphene oxide suspension of 3mg/mL obtains clarifying with 5mL concentration, press then that final concentration 0.12 g/mL adds the NaOH solid and 50 ℃ of stirred in water bath 4 hours, add concentrated hydrochloric acid afterwards and adjust the pH value of solution value near 1, repeated hydrogenation oxygen radical ion not, wash repeatedly and centrifugal purification with distilled water, obtain the water-soluble good not graphite oxide aqueous solution of alkalization.Add silver nitrate pressed powder (difference 18mg, 36mg, 54mg) to graphite oxide aqueous solution (concentration is 0.5mg/ml), adding ultra-pure water to cumulative volume is 100ml, mixing boils, (distinguish 20mg, 40mg is 60mg) as reductant and stabilizing agent to add the trisodium citrate solid again, 100 ℃ are boiled 1h, ultrafiltration post water elution by 10kDa is three times then, removes unconjugated silver-colored particle and other intermediate products, can obtain graphene oxide/silver-colored particle nano-complex.Calculate three kinds of different silver nitrates by complexometry and feed intake that the molar ratio of silver-colored particle and graphene oxide is respectively 0.65,1,2 in the compound of gained.
Above-mentioned graphene oxide/silver-colored particle nano-complex is carried out transmission electron microscope analysis, from Fig. 1, Fig. 2, Fig. 3 as can be known: the silver nano-grain that is combined on the graphene oxide lamella is sphere, the silver-colored amounts of particles difference of combination under the different ratio situations, but size is all between about 10nm ~ 100nm.
With embodiment 1 gained graphene oxide/silver-colored particle nano-complex and bacterial suspension Mixed culture, measure bacterial activity by mtt assay behind the 2.5h.Concrete grammar is: Escherichia coli and staphylococcus aureus are incubated at the LB liquid nutrient medium respectively, shake at constant temperature and be cultured to (OD exponential phase of growth in the window
600Being about 0.5) time gets 200 μ l and is re-seeded into the new medium of 2ml, the GO-Ag nano-complex that adds different volumes, make the compound final concentration of three kinds of different ratios be respectively 2.5,5,10 μ g/ml, put into the constant temperature shaking table and continue to cultivate 2.5h, get this suspension and add 96 orifice plates with every hole 100 μ l, add 20 μ l MTT again, 96 orifice plates are placed 37 ℃ of constant incubator 4h, absorb supernatant, it is fully dissolving crystallized to add 150 μ l dimethyl sulfoxide (DMSO)s again, and microplate reader is being surveyed its absorbance value at 570nm wavelength place.
Referring to Fig. 4, shown in Figure 5: the growth activity of bacterium presents concentration dependent, and material concentration is more high, and bacterial activity is more low; The compound that compares three kinds of different ratios, ratio is that 1 compound shows the strongest anti-microbial property, when its concentration is 10 μ g/ml, the active percentage of Escherichia coli and staphylococcus aureus has only about 20% and 30%, and ratio is that the placed in the middle and ratio of 2 compound anti-microbial property is that 0.65 compound possesses remarkable anti-microbial property hardly.Therefore, above-mentioned ratio is that graphene oxide/silver-colored particle nano-complex of 1 can be used as anti-biotic material efficiently.
Embodiment 3
With embodiment 1 gained graphene oxide/silver-colored particle nano-complex and bacterial suspension Mixed culture, evenly be applied to behind the 2.5h on the solid LB agar plate, by the antibacterial activity of statistics gained bacterium colony number (CFU) analysis of material.Concrete grammar is: Escherichia coli and staphylococcus aureus are incubated at the LB liquid nutrient medium respectively, shake at constant temperature and be cultured to (OD exponential phase of growth in the window
600Being about 0.5) time gets 200 μ l and is re-seeded into the new medium of 2ml, adding ratio is 1 GO-Ag nano-complex, make its final concentration be respectively 2.5,5,10 μ g/ml put into the constant temperature shaking table and continue to cultivate 2.5h, evenly be applied on the solid LB agar plate after the gradient concentration dilution, place 37 ℃ of constant incubators to cultivate 14h, record the bacterium colony number of each planar surface then, the statistics mapping analysis.
Result such as Fig. 6, shown in Figure 7: compare with experimental group, the clump count that the flat board that material processed is crossed grows obviously reduces, and illustrates that this compound can significantly suppress the growth of bacterium.Compare Escherichia coli and staphylococcus aureus again, find when complex concentration is 10 μ g/ml, Escherichia coli are almost all dead, there is not obvious bacterium colony to form on the flat board, staphylococcus aureus then has a small amount of but obvious visible bacterium colony to form, and shows that this graphene oxide/silver-colored particle nano-complex is better than the gram-positive bacteria staphylococcus aureus to the colibacillary inhibition of Gram-negative bacteria.
Embodiment 4
Referring to shown in Figure 8: be that 0.65 compound has no effect to cytoactive with respect to ratio, ratio is that this compound of 1 and 2 has certain influence to cell, when material concentration is 10ug/ml, still there is the cell about 60% to keep its growth activity, with its antibacterial effect comparison efficiently, can think that this compound does not make significant difference substantially to cytoactive, illustrate that the graphene oxide/silver-colored particle nano-complex biological safety of method for preparing is higher.
Claims (2)
1. the preparation method of graphene oxide/silver-colored particle nano-complex is characterized in that, may further comprise the steps:
Step 1) prepares the graphite oxide aqueous solution with improved Hummers method, and the solid sodium chloride of 1g expanded graphite and about 30 times of weight is ground to no obvious visible particle shape thing, adds the water suction filtration, and flush away sodium chloride is with the graphite dry for standby of porphyrize;
The about 30mL concentration of adding is 98.3% the concentrated sulfuric acid in flask, put into the porphyrize graphite of having dried, stirred 8 hours under the room temperature, then mixture is placed ice bath, add about 3g potassium permanganate, slowly be warming up to 115 ℃, slowly add about 150mL water, add about 10mL30% hydrogen peroxide and stirring afterwards, the centrifugal upper water solution of removing is washed to solution repeatedly and no longer occurs till the precipitation;
Be the graphene oxide hydrosol that the ultrasonic 30min of graphene oxide suspension of 3mg/mL obtains clarifying with 5mL concentration, press then that final concentration 0.12 g/mL adds the NaOH solid and 50 ℃ of stirred in water bath 4 hours, add concentrated hydrochloric acid afterwards and adjust the pH value of solution value near 1, repeated hydrogenation oxygen radical ion not, wash repeatedly and centrifugal purification with distilled water, obtain the water-soluble good not graphite oxide aqueous solution of alkalization, step 2) adding the silver nitrate pressed powder in described graphite oxide aqueous solution boils, described silver nitrate and graphene oxide feed intake mass ratio between 7.2 ~ 21.6, step 3) adds trisodium citrate as reductant and stabilizing agent, then again 100 ℃ boil 1h, ultrafiltration pipe wash-out three times, remove unconjugated silver-colored particle and other intermediate products, can obtain graphene oxide/silver-colored particle nano-complex.
2. the application of graphene oxide according to claim 1/silver-colored particle nano-complex, it is antibiotic to it is characterized in that this nano-complex can be used for.
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Cited By (18)
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| CN103521780A (en) * | 2013-11-04 | 2014-01-22 | 中国农业科学院农业质量标准与检测技术研究所 | Preparation method and application of oxidized graphene load gold nanoparticle sol with surface-enhanced Raman spectrum activity |
| CN103975951A (en) * | 2014-04-25 | 2014-08-13 | 常州月夜灯芯绒有限公司 | Graphene oxide / silver complex antibacterial material and preparation method thereof |
| CN104118872A (en) * | 2014-08-04 | 2014-10-29 | 湖南元素密码石墨烯研究院（有限合伙） | Method and device for purifying oxidized graphene/graphene solution |
| CN104353469A (en) * | 2014-10-28 | 2015-02-18 | 江苏大学 | Method for preparing nanocomposite photocatalyst and application of nanocomposite photocatalyst |
| CN104945636A (en) * | 2015-05-13 | 2015-09-30 | 华南师范大学 | Composite material as well as preparation method and use of composite material |
| CN105312063A (en) * | 2014-07-01 | 2016-02-10 | 韩爱英 | Graphene composite catalyst and preparation method thereof |
| CN107337833A (en) * | 2017-07-06 | 2017-11-10 | 湖州中林国际贸易有限公司 | A kind of preparation method for mixing plastic-wood floor material |
| CN107365456A (en) * | 2017-09-15 | 2017-11-21 | 厦门万新橡胶有限公司 | A kind of novel graphite alkene is modified heat conductive rubber |
| CN107617750A (en) * | 2017-09-25 | 2018-01-23 | 中国科学院大连化学物理研究所 | The method for preparing Ag/ redox graphene composite nano materials |
| CN107697905A (en) * | 2017-10-30 | 2018-02-16 | 山西大学 | A kind of preparation method of three-dimensional nitrogen-doped graphene aeroge |
| CN108378440A (en) * | 2018-03-15 | 2018-08-10 | 广西中医药大学 | Chinese medicine antibacterial mask containing graphene oxide-copper silver nanoparticle compound |
| CN108484953A (en) * | 2018-05-09 | 2018-09-04 | 台州骊威环保科技有限公司 | Antibacterial composite membrane of cellulose and its preparation method and application is made with pineapple leaves |
| CN108746580A (en) * | 2018-04-04 | 2018-11-06 | 北京石墨烯技术研究院有限公司 | A kind of dispersing method of graphene oxide in metal powder |
| CN109207984A (en) * | 2018-08-20 | 2019-01-15 | 中国科学院金属研究所 | The preparation method of corrosion resistant antibiotic anti-biofouling multifunctional metal-base protective coating |
| CN109292767A (en) * | 2018-12-07 | 2019-02-01 | 四川聚创石墨烯科技有限公司 | A kind of purification process of graphene oxide |
| CN109292762A (en) * | 2018-12-07 | 2019-02-01 | 四川聚创石墨烯科技有限公司 | A kind of low fault of construction graphene production method |
| CN109292764A (en) * | 2018-12-07 | 2019-02-01 | 四川聚创石墨烯科技有限公司 | A kind of low impurity content graphene preparation method |
| CN117298131A (en) * | 2023-11-29 | 2023-12-29 | 唐宁医药科技(济南)有限公司 | Glycyrrhizic acid-jateorhizine nano-drug for diabetic foot, preparation method and application thereof |
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| CN109292764B (en) * | 2018-12-07 | 2021-01-22 | 四川聚创石墨烯科技有限公司 | Preparation method of low-impurity-content graphene |
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| CN117298131A (en) * | 2023-11-29 | 2023-12-29 | 唐宁医药科技(济南)有限公司 | Glycyrrhizic acid-jateorhizine nano-drug for diabetic foot, preparation method and application thereof |
| CN117298131B (en) * | 2023-11-29 | 2024-02-23 | 唐宁医药科技(济南)有限公司 | Glycyrrhizic acid-jateorhizine nano-drug for diabetic foot, preparation method and application thereof |
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Application publication date: 20130821 |