CN107311143A - A kind of method that utilization Enteromorpha prepares carbon quantum dot - Google Patents
A kind of method that utilization Enteromorpha prepares carbon quantum dot Download PDFInfo
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Abstract
The invention belongs to carbon nanomaterial and ocean technical field of new material preparation, the Enteromorpha being related in a kind of utilization ocean prepares the method for carbon quantum dot, and the carbon quantum dot of preparation is applied into the fields such as metal ion detection and bio-imaging.First Enteromorpha is rinsed well with deionized water and is put into 150ml ptfe autoclaves, then the borate buffer solutions of pH 10.0 are added, reactor is put into vacuum drying chamber again and reacted 6 hours under conditions of 180 DEG C, reactor is then taken out and is cooled to room temperature, obtain carbon quantum dot solution.Obtained carbon quantum dot solution is finally centrifuged into 20min using centrifuge under the conditions of 11900rpm and removes solid impurity and precipitation, it is obtained carbon quantum dot to take supernatant liquor.Its preparation process is simple and convenient to operate, and raw material are easily obtained, and prepares that cost is low, and the carbon quantum dot of preparation is nontoxic, point uniform particle diameter, photoluminescent property are excellent, stability is good.
Description
Technical field：
The invention belongs to carbon nanomaterial and ocean technical field of new material preparation, it is related to the technique that a kind of hydro-thermal method prepares carbon quantum dot, Enteromorpha in particularly a kind of utilization ocean prepares the method for carbon quantum dot, and the carbon quantum dot of preparation is applied into the fields such as metal ion detection and bio-imaging.
Background technology：
Due to reasons such as Global climate change, body eutrophications, a large amount of Enteromorpha green tides in ocean are caused to break out.Prolific Enteromorpha energy shielded from sunlight, influences the growth of seabed algae；Dead Enteromorpha can also consume the oxygen in seawater, and a large amount of Enteromorpha floatings gather bank, block the fairway, while destroying marine ecosystems, seriously threaten inshore fishing, the development of tourist industry.Therefore; the problem of increasingly being paid close attention in terms of the recycling of Enteromorpha and rationally utilization always environmental protection and the utilization of resources; such as there is research worker by Enteromorpha by extruding moisture by mechanical treatment after salvaging; it is then converted into feed and organic fertilizer etc., and explores the processing of more Enteromorphas and recovery and utilization technology still has great importance.
The booming of materialogy promotes many effective nano material synthesis and the appearance of application process.Wherein carbon quantum dot is a kind of new fluorescence carbon nanomaterial, compared with traditional semiconductor-quantum-point, it has that good dispersion in the aqueous solution, chemical stability are strong, is easy to functionalization, good biocompatibility and the characteristic such as photoelectric properties are excellent, has splendid application prospect in fields such as bio-imaging, biochemical sensitive, photoelectrocatalysis.Have much currently used for carbon quantum dot method is prepared, mainly including laser lift-off, arc discharge method, electrochemical stripping method, chemical oxidization method, microwave process for synthesizing and hydro-thermal process method etc.；The carbon source of carbon quantum dot is varied, such as：CNT, carbon fiber, graphite rod, activated carbon etc..But, these methods are more complicated and time-consuming, the raw material of use is mostly costly, and product needs just to can guarantee that the stable optical property of carbon quantum dot by means such as strong acid reflow treatment or surface modifications, limits large-scale production and the practical application of fluorescent carbon quantum dot.Therefore, a kind of raw material cheap and easy to get, Nantural non-toxic is found, quickly can seem particularly necessary by the excellent fluorescent carbon quantum dot of processability using simple effective method.Application No. 201110049051.X Chinese invention patent discloses a kind of fluorescent carbon quantum dot preparation method using plant haulm as raw material, mainly includes the following steps that：High-temperature heating raw material in 350 DEG C -450 DEG C in tubular heater obtain soot；Returned acid is boiled 10-12 hours in salpeter solution；Remove water and obtain solid product；With polyethylene glycol and deionized water are mixed is incorporated in 105 DEG C -135 DEG C and reacts 48-72 hours；Carbon quantum dot is obtained after centrifugation.Although the raw material in the preparation method are cheap and easy to get, processing step is cumbersome, and high to equipment requirement, time-consuming, is unfavorable for large-scale production；The Chinese invention patent of Application No. 201210264411.2 discloses a kind of bean dregs by the use of soybean after squeezing soya-bean milk as the fluorescent carbon quantum dot preparation method of raw material, mainly includes the following steps that：Raw material drying dehydration is obtained into bean dregs powder；20-40min carbonizations are heated in 200 DEG C；Grind the coarse powder that sieves to obtain；It is scattered in deionized water and is concentrated by evaporation；By concentrate in -50 DEG C of freeze-dryings, fluorescent carbon quantum dot is produced.Although the technique of this method is relatively easy, needs to carry out raw material complicated pretreatment, substantially increase production cost.In addition, the granularity uniformity of obtained carbon quantum dot is poor in embodiment, particle size range is wider, about 3-12 μm.Therefore it is related to a kind of method that utilization Enteromorpha prepares carbon quantum dot, preparing raw material is easily obtained and with low cost, while preparation technology is simple, manufacturing cost is low, can be mass-produced, prepared carbon quantum dot uniform particle diameter, application environment is good, and market prospects are boundless.
The content of the invention：
It is an object of the invention to overcome the shortcoming that prior art is present, seek a kind of raw material easily obtain, it is with low cost, preparation technology is simple, preparation process environmental protection, it can be mass-produced out uniform particle diameter, the method for the carbon quantum dot of excellent performance simultaneously, the carbon quantum dot uniform particle diameter prepared using this method, good water solubility and bright blue can be sent under uviol lamp；The carbon quantum of preparation has extremely strong specificity to iron ion detection, can be applied to iron ion in seawater or running water and detect, while may be used on the fields such as bio-imaging.
To achieve these goals, the technique of present invention preparation carbon quantum dot specifically includes following steps：
(1) Enteromorpha that 1-10g is collected into is weighed, is rinsed well with deionized water, is put into 50-200mL ptfe autoclaves, the cushioning liquid of 30-160mL deionized waters or 30-160mL pH for 6.0-10.0 is measured, pours into the reactor.
(2) reactor is put into vacuum drying chamber, 150-240 DEG C, reacts 3-15h, be subsequently cooled to room temperature, obtain carbon quantum dot solution.
(3) the carbon quantum dot solution for preparing step (2) removes impurity and precipitation, it is obtained carbon quantum dot to take supernatant liquor using centrifuge after centrifugation 10-30min under the conditions of 8000-15000rpm.
The present invention is characterized as the carbon quantum dot to obtained by using means of different, iron ion detection and cell imaging can be carried out, the means used are included in fluorescence, ultraviolet, infrared, x-ray photoelectron spectroscopy (XPS) and transmission electron microscope (TEM), AFM (AFM), the one or more of X-ray diffraction (XPD).
Using the Enteromorpha raw material of more than 1g different qualities, in pH value in 1.0-13.0 cushioning liquid, hydrothermal temperature is 150-240 DEG C, and under the 3-15h reaction time, the particle diameter of gained carbon quantum dot is respectively between 2-20nm, and quantum yield is 9-30%；The carbon quantum dot uniform particle sizes of preparation, its surface is rich in groups such as carboxyl, hydroxyls, with splendid water solubility；Gained carbon quantum dot is placed more than 5 months, the fluorescence intensity change very little of the carbon quantum dot sample of same concentrations, and the photoluminescent property of carbon quantum dot is stable；150 μM of Ag are separately added into carbon quantum dot solution+,Al3+,Cd2+,Cu2+,Co2+,Fe2+,Fe3+,Hg2+,Mg2+,Mn2+,Ni2+,Pb2+,Zn2+Etc. different metal ions, wherein Fe3+There is obvious quenching effect to the fluorescence intensity of carbon quantum dot, and with addition Fe3+Concentration increase, the fluorescence intensity of carbon quantum dot constantly reduces, reduction fluorescence intensity and Fe3+It is linear within the specific limits, and the addition for increasing other different metal ions is produced all without the fluorescence intensity to carbon quantum dot and significantly affected, so the carbon quantum dot prepared can be by its fluorescent quenching effect to Fe3+Specific detection.
Prepared carbon quantum dot is applied to cell imaging, significantly, and continuous scanning is more than 10 hours effect effect, and fluorescence intensity is basically unchanged, illustrates that the carbon quantum dot has high anti-light Bleachability.
The present invention compared with prior art, with advantages below：One is that the carbon source for synthesizing carbon quantum dot comes from natural material Enteromorpha, and raw material are easily obtained, and preparation cost is low, also provides new processing path for prolific Enteromorpha in ocean；Two be that the carbon quantum dot prepared is nontoxic, putting uniform particle diameter, photoluminescent property, excellent, stability is good；Three be that preparation technology is simple to operate, preparation efficiency is high, can be mass-produced, while environmental protection.
Brief description of the drawings：
The TEM figures and high-resolution for the carbon quantum dot that Fig. 1 is prepared for the present invention transmit electric (HRTEM) figure.
Fig. 2 is carbon quantum dot size distribution plot.
Fig. 3 is carbon quantum dot atomic force microscopy diagram
Fig. 4 is the respective heights figure of carbon quantum dot atomic force microscopy diagram.
Enter the Fe of various concentrations in the carbon quantum dot that Fig. 5 is prepared for the present invention in the embodiment 2 of same concentration3+Fluorescence intensity afterwards.
Fig. 6 uses preparation carbon quantum dot detection Fe in implementing 2 for the present invention3+Linear relationship, with the Fe that various concentrations are added in the carbon quantum dot of concentration3+Afterwards, fluorescence intensity (Y) and Fe3+The linear relationship of concentration (X), (1) is Y=953.05-1.76X (R=0.9971), 1-350 μM of concentration range；(2) it is Y=410.46-0.173X (R=0.9933), 370-1700 μM of concentration range.
Fig. 7 is the laser scanning co-focusing microscope imaging fluorogram for the HeLa cells that the embodiment of the present invention 3 is incubated through carbon quantum dot solution
Fig. 8 is the laser scanning co-focusing microscope imaging light field figure for the HeLa cells that the embodiment of the present invention 3 is incubated through carbon quantum dot solution
Fig. 9 is the laser scanning co-focusing microscope imaging light field figure for the HeLa cells that the embodiment of the present invention 3 is incubated through carbon quantum dot solution and the overlay chart of fluorogram
Figure 10 is the cytotoxicity test result figure of carbon quantum dot
Embodiment：
Below by embodiment and the invention will be further described with reference to accompanying drawing.
Embodiment 1：
(1) Enteromorpha that 2g is collected into is weighed, is rinsed well with deionized water, is put into 150ml ptfe autoclaves, the borate buffer solutions of 120ml pH 10.0 are then added.
(2) reactor is put into vacuum drying chamber and reacted 6 hours under conditions of 180 DEG C, then taken out reactor and be cooled to room temperature, obtain carbon quantum dot solution.
(3) the carbon quantum dot solution for preparing step (2) centrifuges 20min using centrifuge under the conditions of 11900rpm and removes solid impurity and precipitation, and it is obtained carbon quantum dot to take supernatant liquor.
Embodiment 2：
Carbon quantum dot solution prepared by embodiment 1 is applied to Fe by the present embodiment3+The context of detection of characteristic ion, is separately added into 150 μM of Ag+,Al3+,Cd2+,Cu2+,Co2+,Fe2+,Fe3+,Hg2+,Mg2+,Mn2+,Ni2+,Pb2+,Zn2+, and its fluorescence intensity is surveyed under 360nm excitation wavelength, wherein only Fe3+There is obvious quenching effect to the fluorescence intensity of carbon quantum dot, the addition of other different metal ions is produced all without the fluorescence intensity to carbon quantum dot to be significantly affected.In order to probe into the Fe of addition3+The linear relationship of concentration and fluorescence intensity, prepares a series of Fe of concentration gradients3+, 1 μM, 10 μM, 50 μM, 100 μM, 120 μM, 150 μM, 170 μM, 200 μM, 220 μM, 250 μM, 270 μM, 300 μM, 320 μM, 350 μM, 370 μM, 400 μM, 500 μM, 700 μM, 1000 μM, 1200 μM, 1500 μM, 1700 μM, 2000 μM are added separately in the carbon quantum dot solution of same concentration, a series of different fluorescence intensities are obtained, so as to obtain Fe3+Influence to the fluorescence intensity of carbon quantum dot, as shown in figure 5, the carbon quantum dot prepared using embodiment 1 can realize that lowest detection is limited to 1 μM of Fe3+Detection, Fig. 6 is the different Fe of addition3+, fluorescence intensity (Y) and Fe3+The linear relationship of concentration (X), (1) is Y=953.05-1.76X (R=0.9971), 1-350 μM of concentration range；(2) it is Y=410.46-0.173X (R=0.9933), 370-1700 μM of concentration range.
Embodiment 3：
In terms of carbon quantum dot solution prepared by embodiment 1 to be applied to the present embodiment cell imaging, first by HeLa cells (106Cell/sample) it is placed in length of side 35mm square-shaped slide, the fresh DMEM solution of carbon containing quantum dot solution concentration 1/10 is prepared, being incubated HeLa cells 4-24h with the DMEM solution prepared at 37 DEG C obtains incubated cell；Again at room temperature, all incubated cells are flushed three times with phosphate buffer solution and remove unnecessary unabsorbed carbon quantum dot, selection 488nm Ar ion lasers are excited to incubated cell, and incubated cell is obtained into image by the collection of LEICA TCS SP2 laser scanning co-focusing microscopes, and further collection changes over time the cell imaging situation of a certain FX, as shown in fig. 7, visible cell imaging effect is very good.
The present embodiment is tested cytotoxicity, cell is carried out with the DMEM solution of the carbon quantum dot containing various concentrations to be incubated 24 hours, then the MTT solution that 10mL contains 5mg/mL is added, then proceed to be incubated 4 hours under the conditions of logical 5%CO2 and 37 DEG C, finally crystal is set fully to dissolve with 100 μ L dimethyl sulfoxide (DMSO)s, the light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm, measurement result is as shown in Figure 10, the cytoactive being incubated without carbon quantum dot solution is set as 100%, after then the carbon quantum dot through various concentrations is incubated, cytoactive can keep more than 85%, fluorescence intensity changes over time smaller simultaneously.
Carbon quantum dot is imaged by the present embodiment applied to effective cell, experimental result illustrates that the carbon quantum dot produced successfully can be absorbed by cell, the good spectral characteristic of carbon quantum dot may be used as effective bio-imaging reagent, while the carbon quantum dot has low toxicity, stabilization and influenceed less characteristic by photobleaching.
Claims (6)
1. a kind of method that utilization Enteromorpha prepares carbon quantum dot, it is characterised in that specifically include with
Lower step：
(1) Enteromorpha that 1-10g is collected into is weighed, is rinsed well, is put into deionized water
In 50-200mL ptfe autoclaves, measure 30-160mL deionized waters or
30-160mL pH are 6.0-10.0 cushioning liquid, are poured into the reactor.
(2) reactor is put into vacuum drying chamber, 150-240 DEG C, reacts 3-15h,
Room temperature is subsequently cooled to, carbon quantum dot solution is obtained.
(3) the carbon quantum dot solution for preparing step (2) uses centrifuge in 8000-15000
Centrifuged under the conditions of rpm after 10-30min, remove impurity and precipitation, it is system to take supernatant liquor
The carbon quantum dot obtained.
2. the method for carbon quantum dot, its feature are prepared using Enteromorpha according to claim 1
It is for the raw material for preparing carbon quantum dot from Enteromorpha common in the sea.
3. preparing the method for carbon quantum dot using Enteromorpha according to claim 1, its feature exists
In preparation carbon quantum dot particle diameter respectively between 2-20nm, quantum yield is 9%-30%；
The carbon quantum dot uniform particle sizes of preparation, its surface is rich in groups such as carboxyl, hydroxyls, with splendid
Water solubility.
4. the method for carbon quantum dot, its feature are prepared using Enteromorpha according to claim 1
It is that the carbon quantum dot photoluminescent property prepared is stable, can be by its fluorescent quenching effect to Fe3+It is special
Opposite sex detection.
5. the method for carbon quantum dot, its feature are prepared using Enteromorpha according to claim 1
It is that the carbon quantum dot prepared has high anti-light Bleachability, continuous scanning is more than 10 hours,
Fluorescence intensity is basically unchanged, and can be used in cell imaging.
6. preparing the method for carbon quantum dot using Enteromorpha according to claim 1, its feature exists
In the carbon quantum dot safety and low toxicity of preparation, by DMEM of the cell with the carbon quantum dot containing various concentrations
Solution be incubated 24 hours, then adds the MTT solution that 10mL contains 5mg/mL,
Then proceed to be incubated 4 hours under the conditions of logical 5%CO2 and 37 DEG C, finally with 100 μ L
Dimethyl sulfoxide (DMSO) makes crystal fully dissolve, and measures each at enzyme-linked immunosorbent assay instrument 490nm
The light absorption value in hole, the cytoactive being incubated without carbon quantum dot solution is set as 100%, then
After carbon quantum dot through various concentrations is incubated, cytoactive can keep more than 85%, while fluorescence
Intensity changes over time smaller.
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| CN110155984A (en) * | 2019-06-12 | 2019-08-23 | 浙江科技学院 | Using soybean slag as the method and application of raw material hydro-thermal method synthesising biological matter fluorescent carbon point |
| CN111234815A (en) * | 2020-01-22 | 2020-06-05 | 哈尔滨工业大学 | Preparation and use methods of biomass carbon quantum dot fluorescence detector |
| CN111621292A (en) * | 2020-06-28 | 2020-09-04 | 浙江工业大学 | Preparation method of large marine plant-based carbon quantum dots |
| CN112574743A (en) * | 2020-12-14 | 2021-03-30 | 广西大学 | One-step hydrothermal synthesis method of multi-doped carbon quantum dots |
| CN113003562A (en) * | 2021-02-05 | 2021-06-22 | 广东粤港澳大湾区国家纳米科技创新研究院 | Carbon nano-particle and preparation method and application thereof |
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| CN112574743A (en) * | 2020-12-14 | 2021-03-30 | 广西大学 | One-step hydrothermal synthesis method of multi-doped carbon quantum dots |
| CN113003562A (en) * | 2021-02-05 | 2021-06-22 | 广东粤港澳大湾区国家纳米科技创新研究院 | Carbon nano-particle and preparation method and application thereof |
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