CN108144059A - A kind of two dimension black phosphorus nanometer sheet loading nano silvery composite material and preparation method thereof and the application as antibacterial agent - Google Patents
A kind of two dimension black phosphorus nanometer sheet loading nano silvery composite material and preparation method thereof and the application as antibacterial agent Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material, this method is to add in soluble silver salt in black phosphorus nanometer sheet dispersion liquid, it is ultrasonically treated, up to two-dimentional black phosphorus nanometer sheet loading nano silvery composite material, this method is to realize black phosphorus nanometer sheet to nano silver in-situ reducing and load by one kettle way under room temperature, normal pressure, prepare nano silver be uniformly dispersed, the two-dimentional black phosphorus nanometer sheet loading nano silvery composite material of load stabilization；The composite material can be used as antibacterial agent, particularly have better inhibition effect to the activity of Escherichia coli and staphylococcus aureus.
Description
Technical field
The present invention relates to a kind of antibacterial reagents, and in particular to a kind of black phosphorus nanometer sheet loading nano silvery composite material and one
The method and black phosphorus nanometer sheet loading nano silvery of the reduction of pot method ultrasonic wave added black phosphorus nanometer sheet and loading nano silvery in situ are compound
Application of the material as antibacterial reagent, belongs to biological medicine field of material preparation.
Background technology
Black phosphorus nanometer sheet, as a kind of New Two Dimensional nano material, since its special structure and performance are closed extensively
Note.Black phosphorus plays what rennet was formed by phosphorus, and interlayer Van der Waals force is weaker, so can be surpassed by simple ultrasonic stripping method
Thin two-dimensional nano piece, since the crystalline structure of its two-dimensional appearance is similar to graphene, be otherwise known as phosphorus alkene.The band gap of black phosphorus with
Its thickness is related, and the band gap of bulk black phosphorus is about 0.3eV, and the band gap of individual layer black phosphorus nanometer sheet is then 2.0eV.Since black phosphorus is received
Rice piece has high electron mobility, and in-plane anisotropy and the absorption band across ultraviolet and entire visible light region etc. are unique
Property makes it be widely studied in electricity, photoelectricity, sensor and biological medicine etc..Black phosphorus nanometer sheet is cured in biology
Prescription face has tremendous potential, and optical dynamic therapy (PDT) reagent, photo-thermal therapy (PTT) reagent can be used as and resist swollen by being studied
Tumor medicine carrier.
Nano silver has extensive use in fields such as catalysis, energy storage and biological medicines.However, simple nano silver holds
Easily assemble, performance comes with some shortcomings, and generally all by being supported in other nano materials, can improve nano silver
Dispersion stabilization, obtain the higher composite material of comprehensive performance.Two-dimensional material has high-specific surface area, can load well
Nano silver however, current most of two-dimensional material does not all have reduction activation in itself, needs additionally to add reducing agent and stabilization
Agent could realize the uniform load to nano silver.
The report of the loading nano silvery in the black squama nanometer sheet of two dimension, Chinese patent (publication number are had been reported that in the prior art
CN106623967A a kind of black squama-metal nanometer composite material and its synthetic method and application) are disclosed, it is specific open：(1) it takes
Black phosphorus BP crystal is scattered in N-Methyl pyrrolidone, then carries out break process, obtains broken liquid；(2) broken liquid is surpassed
Sonication, centrifugation, takes precipitation, precipitation is resuspended in N-Methyl pyrrolidone, centrifuge again, collects supernatant, obtains BP nanometer sheets
Suspension；(3) it in the water for boiling the addition of BP nanometer sheets suspension, is subsequently added into chlorauric acid solution or silver nitrate solution continues boiling instead
Should, it cools down, centrifuges after reaction, collect precipitation, obtain black phosphorus-metal nanometer composite material.This method using BP in itself also
The metal ion adsorbed on BP surfaces is reduced into metal element by originality, generates black phosphorus-metal nano compound, easy, easy behaviour
Make, manufactured black phosphorus-metal nanometer composite material can be applied in terms of photo-thermal effect and be increased as active substrate on surface
Application in strong Raman scattering SERS.This method needs react under 100 DEG C of hot conditions, and reaction condition requirement is high, particularly
Easily make black squama excessive oxidation under the high temperature conditions, composite property is caused to be deteriorated.
At present, in terms of black phosphorus-metal nanometer composite material of report is mainly used for photo-thermal effect and as active substrate
Application in Surface enhanced Raman scattering SERS, but in terms of sterilization and have not been reported.
Invention content
For solve in the prior art two-dimentional black squama loading nano silvery method there are the defects of, of the invention first purpose
It is to be to provide a kind of side under room temperature, normal pressure by one kettle way realization black phosphorus nanometer sheet to nano silver in-situ reducing and load
Method.
Another object of the present invention be to provide a kind of nano silver be uniformly dispersed, the two-dimentional black phosphorus nanometer of load stabilization
Piece loading nano silvery composite material.
Third object of the present invention is to be to provide a kind of answering for two-dimentional black phosphorus nanometer sheet loading nano silvery composite material
With as the application of antibacterial agent, being answered especially as the reagent of Escherichia coli and staphylococcus aureus activity is inhibited
With.
In order to realize above-mentioned technical purpose, the present invention provides a kind of two-dimentional black phosphorus nanometer sheet loading nano silvery composite materials
The preparation method of (Ag@BP), the preparation method are that soluble silver salt is added in black phosphorus nanometer sheet dispersion liquid, are ultrasonically treated, i.e.,
.
Preferred scheme, a concentration of 0.05~5mg/mL of black squama nanometer sheet in the black phosphorus nanometer sheet dispersion liquid.It is preferred that
Scheme, solvent is at least one of water, methanol and ethyl alcohol in the black phosphorus nanometer sheet dispersion liquid.Preferred solvent is water.
Preferred scheme, the soluble silver salt are silver nitrate and/or silver perchlorate.
Preferred scheme, a concentration of 1~100mM of addition of the soluble silver salt in black phosphorus nanometer sheet dispersion liquid.
Preferred scheme, the frequency of the supersound process are 20~40kHz, and power is 150~300W, the time for 0.25~
3h。
Preferred scheme, the thickness of the black phosphorus nanometer sheet is 1~7nm, a diameter of 200nm~250nm.
The present invention also provides a kind of two-dimentional black phosphorus nanometer sheet loading nano silvery composite materials, are obtained by above-mentioned preparation method
It arrives.
The present invention also provides a kind of application of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material, as antibacterial
Reagent application.
Preferred scheme under the conditions of near infrared light photograph, is pressed down using two-dimentional black phosphorus nanometer sheet loading nano silvery composite material
Bacterial activity processed.
Preferred scheme, using two-dimentional black phosphorus nanometer sheet loading nano silvery composite material as inhibition Escherichia coli and golden yellow
The reagent application of activity against staphylococci.
The preparation method of the black phosphorus nanometer sheet of the present invention：Black phosphorus powder is placed in the saturation sodium hydroxide solution of NMP, ice
Ultrasound stripping under the conditions of bath, then again through centrifuging, washing, again moisture dissipate (or alcohol disperses) the black phosphorus nanometer sheet dispersion
Liquid；Wherein, ultrasonic 20~40kHz of frequency, ultrasonic power are 150~300W, and sonication treatment time is 6~12h.
The preparation method of the black phosphorus nanometer sheet loading nano silvery of the present invention：It is added in black phosphorus nanometer sheet dispersion liquid soluble
Silver salt is ultrasonically treated, centrifugation, washing, it is dry to get；Black squama nanometer sheet is a concentration of in the black phosphorus nanometer sheet dispersion liquid
0.05~5mg/mL；A concentration of 1~100mM of addition of the soluble silver salt in black phosphorus nanometer sheet dispersion liquid；The supersound process
Time for 0.25~3h, ultrasonic frequency is 20~40kHz, and ultrasonic power is 150~300W.
Technical scheme of the present invention makes full use of the oxidation-reduction potential of black phosphorus to be less than the oxidation-reduction potential of silver ion
The characteristics of, black phosphorus may be used, silver ion is restored.Silver ion is restored using black phosphorus nanometer sheet surface, is realized
Silver-colored in-situ reducing and load, and black phosphorus surface is steady well since a small amount of phosphorous oxides or phosphoric acid of oxidation generation have nano silver
It is set for using, it is ensured that nano silver is in the homodisperse of black phosphorus surface.
Technical scheme of the present invention, there are large effect, black phosphorus nanometer sheets for reaction of the ultrasound to black phosphorus reduction silver ion
After being mixed with silver ion, do not need to ultrasound, black phosphorus nanometer sheet surface can also generate nano silver, but the nano silver size generated compared with
Greatly, it is and uneven, and the nano silver size obtained under ultrasound condition is smaller and uniform, can greatly improve the activity of silver；Ultrasound
Dispersion of the silver ion on black phosphorus nanometer sheet surface can be advantageously promoted, and ultrasound is a kind of uniform energy injection mode, favorably
In the quick formation of nano silver, the aggregation of nano-particle is reduced, ensure that nano silver in the uniformly dispersed of black phosphorus surface.
Technical scheme of the present invention is compound by two-dimentional black phosphorus nanometer sheet and nano silver, can greatly improve the two in terms of sterilization
Synergistic effect, on the one hand can utilize black phosphorus photo-thermal effect, make local temperature excessively high, lead to bacterium internal protein and lipid
A large amount of bacteriums are killed in denaturation, the metabolism so as to inhibit bacteria in a short time；On the other hand, Ag@BP also can slow release
Go out silver ion, destroy the cell membrane of bacterium and improve the ROS contents in bacterium, the proliferation so as to inhibit bacteria, the two collaboration is made
With apparent, bactericidal effect is excellent.
Technical scheme of the present invention is used by the use of nano silver/black phosphorus nanometer sheet (Ag@BP) composite material as anti-biotic material
Method be：A certain amount of Ag@BP are dispersed in PBS buffer solution (pH=7.4), and be added separately to give birth to Escherichia coli and
In the culture medium of staphylococcus aureus, after cultivating 4h, with 808nm, 2.5Wcm-2Laser irradiation 5min, continue cultivate for 24 hours after
Its OD value is tested by microplate reader and calculates Survival probability of bacteria.The further antibacterial effect of research Ag@BP, using plate count
Method carries out antibacterial activity experiment, after above-mentioned inoculum is diluted 100 times, 50 μ L is taken to be applied on solid medium, 37 DEG C
Lower cultivation is for 24 hours.
Compared with the prior art, the advantageous effects brought of technical scheme of the present invention：
(1) preparation method for preparing black phosphorus nanometer sheet loading nano silvery composite material of the invention is simple, quick, economical,
It can realize that black phosphorus nanometer sheet to one pot of in-situ reducing of nano silver and load, effectively prevent the excessive oxygen of black phosphorus in a mild condition
Change, black phosphorus is both support substrate and stabilizer or silver ion reducing agent.
(2) Ag@BP of the invention are with excellent photostability and photothermal conversion performance, the bacterium being processed,
After near-infrared laser irradiation, almost all is dead, has extremely strong antibacterial ability.
(3) Ag BP of the invention almost no cytotoxicities, have good biological safety.
(4) Ag@BP preparation methods of the invention are simple, and reaction condition is mild, at low cost, and application prospect is big.
Description of the drawings
【Fig. 1】Scheme for embodiment with the synthesis schematic diagram of comparative example 1 and corresponding TEM.As seen from Figure 1：Ultrasonic energy
Dispersion of the silver ion on black phosphorus nanometer sheet surface is advantageously promoted, reduces the aggregation of nano-particle, ensure that nano silver in black phosphorus
Surface it is uniformly dispersed.
【Fig. 2】A) the Antibacterial Mechanism schematic diagram for nano silver/black phosphorus nanometer sheet (Ag@BP) under near-infrared laser irradiation,
On the one hand the photo-thermal effect of black phosphorus can be utilized, makes local temperature excessively high, kills a large amount of bacteriums in a short time, on the other hand,
Ag@BP can also slowly release silver ion, the proliferation inhibited bacteria；B) it is Ag@BP, BP and the photo-thermal effect comparison diagram of pure water,
As can be seen that Ag@BP maintain the photothermal conversion ability of BP well；Ag@BP swash in 808nm near-infrareds it can be seen from c)
There is good photostability under light；Figure is d) growth curve of bacteria under the conditions of different disposal, wherein (1) is blank control,
(2) it is irradiated for only near-infrared laser, (3) only to blacken phosphorus nanometer sheet (corresponding comparative example 2), (4) is only add Ag@BP (corresponding
Comparative example 3), (5) are black phosphorus nanometer sheet+near-infrared laser irradiation (corresponding comparative example 4), and (6) are shone for Ag@BP+ near-infrared lasers
It penetrates (corresponding embodiment)；E) and f) it is respectively the photo and quantitative analysis of bacterium colony on corresponding agar plate；By d), e) and f) may be used
To find out, through the processed bacteriums of Ag@BP, after near-infrared laser irradiation, almost all is dead, has extremely strong antibacterial energy
Power.
Specific embodiment
With reference to specific embodiment, the invention will be further described.These embodiments are interpreted as being merely to illustrate
The present invention rather than limit the scope of the invention.After the content of the invention recorded has been read, based on the present invention's
Principle equally falls into claims of the present invention and is limited to the various changes of the invention made or modification
Embodiment 1
The preparation of step (1) BP nanometer sheets：
Black phosphorus powder sample prepares ultra-thin BP nanometer sheets by the stripping of ultrasonic stripping method.Detailed process is as follows：Take 15mg
Bulk black phosphorus powder is dispersed in N-Methyl pyrrolidone (NMP) solution of 30mL saturations NaOH, and ultrasound 8h, obtains under condition of ice bath
Brown suspension, subsequent low-speed centrifugal (3000r/min) 10min to remove unstripped black phosphorus residue, take supernatant up to palm fibre
Color black phosphorus nanometer sheet transparency liquid.Acquired solution is stored in 4 DEG C of refrigerators for use, is centrifuged using preceding high speed (12000r/min)
10min obtains black phosphorus nanometer pellet, and ultrasonic disperse is in required solution.
The preparation of step (2) nano silver/black phosphorus nanometer sheet (Ag@BP)：
AgNO3 (100 μ L 10mM) is added in the aqueous solution (1mL 0.10mg/mL) of black phosphorus nanometer sheet, mixed liquor is surpassed
Sound 30 minutes, 12000 leave the heart 10 minutes, and with pure water for several times
The test of antibacterial effect：
A certain amount of Ag@BP are dispersed in PBS buffer solution (pH=7.4), and be added separately to give birth to Escherichia coli and
In the culture medium of staphylococcus aureus, after cultivating 4h, with 808nm, 2.5Wcm-2Laser irradiation 5min, continue cultivate for 24 hours after
Its OD value is tested by microplate reader and calculates Survival probability of bacteria.The further antibacterial effect of research Ag@BP, using plate count
Method carries out antibacterial activity experiment, after above-mentioned inoculum is diluted 100 times, 50 μ L is taken to be applied on solid medium, 37 DEG C
Lower cultivation is for 24 hours.
Comparative example 1
The preparation of step (1) BP nanometer sheets：
Black phosphorus powder sample prepares ultra-thin BP nanometer sheets by the stripping of ultrasonic stripping method.Detailed process is as follows：Take 15mg
Bulk black phosphorus powder is dispersed in N-Methyl pyrrolidone (NMP) solution of 30mL saturations NaOH, and ultrasound 8h, obtains under condition of ice bath
Brown suspension, subsequent low-speed centrifugal (3000r/min) 10min to remove unstripped black phosphorus residue, take supernatant up to palm fibre
Color black phosphorus nanometer sheet transparency liquid.Acquired solution is stored in 4 DEG C of refrigerators for use, is centrifuged using preceding high speed (12000r/min)
10min obtains black phosphorus nanometer pellet, and ultrasonic disperse is in required solution.
Preparation of the step (2) without nano silver under ultrasound condition/black phosphorus nanometer sheet (Ag@BP)：
AgNO3 (100 μ L 10mM) is added in the aqueous solution (1mL 0.10mg/mL) of black phosphorus nanometer sheet, mixed liquor is put
It puts 30 minutes, 12000 leave the heart 10 minutes, and with pure water for several times
Comparative example 2
The preparation of black phosphorus nanometer sheet is the same as 1 step of embodiment (1)
The test of antibacterial effect：
A certain amount of BP is dispersed in PBS buffer solution (pH=7.4), and is added separately to give birth to Escherichia coli and golden yellow
In the staphylococcic culture medium of color, after cultivating 4h, continue to cultivate for 24 hours, testing its OD value by microplate reader calculates Survival probability of bacteria.
The antibacterial effect of BP is further studied, antibacterial activity experiment is carried out using the method for plate culture count, above-mentioned inoculum is dilute
After releasing 100 times, 50 μ L is taken to be applied on solid medium, are cultivated for 24 hours at 37 DEG C.
Comparative example 3
The preparation of nano silver/black phosphorus nanometer sheet (Ag@BP) is the same as embodiment 1
The test of antibacterial effect：
A certain amount of Ag@BP are dispersed in PBS buffer solution (pH=7.4), and be added separately to give birth to Escherichia coli and
In the culture medium of staphylococcus aureus, after cultivating 4h, continue to cultivate for 24 hours, its OD value calculating bacterium is tested by microplate reader and is deposited
Motility rate.The further antibacterial effect of research Ag@BP carries out antibacterial activity experiment, by above-mentioned bacterium using the method for plate culture count
After culture solution dilutes 100 times, 50 μ L is taken to be applied on solid medium, are cultivated for 24 hours at 37 DEG C.
Comparative example 4
The preparation of black phosphorus nanometer sheet is the same as 1 step of embodiment (1)
The test of antibacterial effect：
A certain amount of BP is dispersed in PBS buffer solution (pH=7.4), and is added separately to give birth to Escherichia coli and golden yellow
In the staphylococcic culture medium of color, after cultivating 4h, with 808nm, 2.5Wcm-2Laser irradiation 5min, continue cultivate pass through afterwards for 24 hours
Microplate reader tests its OD value and calculates Survival probability of bacteria.The antibacterial effect of BP is further studied, is carried out using the method for plate culture count
Antibacterial activity is tested, and after above-mentioned inoculum is diluted 100 times, 50 μ L is taken to be applied on solid medium, are cultivated at 37 DEG C
24h。
Claims (10)
1. a kind of preparation method of two dimension black phosphorus nanometer sheet loading nano silvery composite material, it is characterised in that：In black phosphorus nanometer sheet
Soluble silver salt is added in dispersion liquid, be ultrasonically treated to get.
2. a kind of preparation method of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material according to claim 1, special
Sign is：A concentration of 0.05~5mg/mL of black squama nanometer sheet in the black phosphorus nanometer sheet dispersion liquid.
3. a kind of preparation method of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material according to claim 1 or 2,
It is characterized in that：Solvent is at least one of water, methanol and ethyl alcohol in the black phosphorus nanometer sheet dispersion liquid.
4. a kind of preparation method of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material according to claim 1, special
Sign is：The soluble silver salt is silver nitrate and/or silver perchlorate.
5. a kind of preparation method of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material according to claim 1, special
Sign is：A concentration of 1~100mM of addition of the soluble silver salt in black phosphorus nanometer sheet dispersion liquid.
6. a kind of preparation side of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material according to claim 1,2,4 or 5
Method, it is characterised in that：The frequency of the supersound process is 20~40kHz, and power is 150~300W, and the time is 0.25~3h.
7. a kind of preparation method of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material according to claim 1, special
Sign is：The thickness of the black phosphorus nanometer sheet is 1~7nm, a diameter of 200nm~250nm.
8. a kind of two dimension black phosphorus nanometer sheet loading nano silvery composite material, it is characterised in that：It is made by any one of claim 1~7
Preparation Method obtains.
9. a kind of application of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material according to any one of claims 8, it is characterised in that：Make
For antibacterial agent application.
10. a kind of application of two-dimentional black phosphorus nanometer sheet loading nano silvery composite material according to claim 9, feature exist
In：Under the conditions of near infrared light photograph, activity is inhibited bacteria using two-dimentional black phosphorus nanometer sheet loading nano silvery composite material.
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