CN106629659A - Preparation method of fluorescent carbon quantum dots employing algae as carbon sources and application - Google Patents
Preparation method of fluorescent carbon quantum dots employing algae as carbon sources and application Download PDFInfo
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Abstract
The invention discloses a method for preparing fluorescent carbon quantum dots employing algae as raw materials. The method comprises the following steps of (1) cleaning the alga raw materials to remove impurities, and crushing the algae and water to obtain alga slurry; (2) putting a proper amount of alga slurry into a sealing system for high-temperature hydrothermal reaction, and carbonizing the alga slurry; (3) carrying out post-treatment on the carbonized product in the step (2) to remove insoluble substances and large-size components, thereby obtaining a fluorescent carbon quantum dot solution; and (4) carrying out freeze drying on the fluorescent carbon quantum dot solution in the step (3) to obtain fluorescent carbon quantum dot powder. Cheap algae, especially enteromorpha prolifera, are effectively converted into the carbon quantum dots with fluorescence characteristics, and high additional values and harmless conversion of the enteromorpha prolifera are achieved; and meanwhile, the preparation method is simple in process, high in operability and low in cost, massive preparation of the carbon quantum dots can be achieved, and other chemicals are not needed, so that the method is environment-friendly; and the yield of a product is high in the preparation process.
Description
Technical field
The invention belongs to functional nanomaterials technical field, and in particular to a kind of carbon quantum dot with fluorescent characteristic
Preparation method, more particularly to a kind of side for preparing the carbon quantum dot cell marking probe with fluorescent characteristic as carbon source with marine alga
Method.
Background technology
Functional nanomaterials are to develop a very fast technical field in recent years, and scientists are successfully made
It is standby go out including organic polymer bead, liposome, semiconductor-quantum-point, silicon quantum dot, upper conversion nano-probe, noble metal nano
Particle, magnetic Nano bead etc. have developed various in interior various nano materials using their various excellent properties
Technological means, it is widely used in each fields such as medical science detection, medical diagnosis on disease, energy battery, environmental assessment, pollution controls.
Carbon quantum dot is one new study hotspot of field of nanometer material technology.It refers to carbon of the diameter in 2-40 nanometer ranges
Atom microballoon.Carbon quantum dot shows significant fluorescence special while material with carbon element various excellent physical and chemical performances are retained
Property and good aqueous phase dispersibility.Experiment shows that carbon quantum dot can be each directly as a kind of safely and effectively fluorescence probe
Kind of cell internalizing is so as to carry out fluorescence labeling to these cells, and the carbon quantum dot of surface modification being capable of specifically marked tumor
Cell.In a word, carbon quantum dot has important using value in the field such as biological detection and bioanalysis.
The preparation method of carbon quantum dot mainly has two big class, and a class is to utilize high purity graphite, carbon black and CNT for original
Material is broken using the method such as beamwriter lithography erosion, electrochemical oxidation, chemical stripping, microwave and ultrasonic wave added, oxygen plasma treatment
Interaction force between bad graphite flake layer, while forming substantial amounts of oxide group in lamella, graphene sheet layer is cut open
Obtain undersized carbon quantum dot；An other class is to utilize the micromolecular compound such as citric acid and ethylenediamine, at a certain temperature
Condensation reaction is carried out, the carbon quantum dot with higher molecular weight is generated.The raw material of both preparation methods is all highly purifiedization
Compound, it is relatively costly.And can usually use red fuming nitric acid (RFNA) and the concentrated sulfuric acid, the harmless treatment of byproduct of reaction during synthesizing
It is complex.In recent years, scientist is found that the method that various natural organic matters prepare carbon quantum dot, such as Chinese patent
CN2014108419973 discloses a kind of method of the carbon quantum dot fluorescence labeling material with egg as carbon source, Chinese patent
A kind of preparation methods of photic carbon quantum dot of giving out light of CN2012102644112 are disclosed and adopt soya-bean milk residue for raw material preparation carbon amounts
The method of son point, Chinese patent 2013102199000 discloses the method for preparing carbon quantum dot as raw material with plant leaf, but
Generally existing product yield is low in these natural organic matter preparation process, such as adopts soya-bean milk residual product yield for 5-7.3%, and
During be related to the use of the chemicals such as sulfuric acid, NaOH, be not belonging to green production process, in addition these raw material are still
So have price higher, the shortcoming of limited source.
Our national cultivated land resources are limited, but possess wide ocean area, and the yield of marine alga is huge, can be used as one
Plant the high biomass resource of valency low amounts.Enteromorpha is one kind of large-scale wild marine alga, strong with fertility, and environment adapts to model
The characteristics of enclosing wide.Recently as a series of change of environmental factors such as global air temperature warming, body eutrophication, ocean Enteromorpha
Green tide is continuously broken out.Most just there is Enteromorpha earlier than 2007 in Yellow Sea of China, continuous 9 years so far outburst Enteromorpha green tide disasters.
Such as in June, 2015 had larger area Enteromorpha to float in Along The Coast of Shandong Province marine site, about 430 square kilometres of green tide area coverage, distribution
About 35200 square kilometres of area (www.xinhuanet.com's news).A large amount of Enteromorpha floatings are gathered in sea, block the passage of ship's navigation.
With this simultaneously, the raised growth propagation of Enteromorpha and the process of decomposition of rotting are required for consuming a large amount of oxygen, and to it marine site is grown
Other hydrobiological existence constitute a serious threat.The Enteromorpha that cannot be removed in time gives out a foul smell when rotting, and affects coastal trip
Trip industry development.How harmless treatment is carried out to Enteromorpha, be the severe challenge that we face by its ' turning waste into wealth '.At present
It is to be made into feed to the harmless treatment main path of Enteromorpha, or processing and eating.
Based on discussed above, design and realize effective approach, Enteromorpha is converted into functional fluorescent carbon quantum dot
Material, conversion harmless for the high added value of Enteromorpha has important practical significance.
The content of the invention
Present invention aims to the deficiency in existing carbon quantum dot synthetic technology, there is provided one kind is with marine alga as raw material
Production fluorescent carbon quantum dot and the method being applied as cell marking probe are prepared, marine alga, particularly Enteromorpha is solved high
The problem of added value harmless treatment.
In order to realize above-mentioned technical proposal, the preparation side of the fluorescent carbon quantum dot with marine alga as carbon source according to the present invention
Method, specifically includes following steps：
(1) marine alga raw material is cleaned to go the removal of impurity, then together crush marine alga and water, obtain seaweed slurry；
(2) appropriate seaweed slurry is put in sealing system carries out high temperature hydro-thermal reaction and makes marine alga slurry be carbonized；
(3) carbonized product in step (2) is post-processed to remove insoluble matter and large-sized component, is obtained fluorescence
Carbon quantum dot solution；
(4) the fluorescent carbon quantum dot solution in step (3) is carried out into freeze-drying process, obtains fluorescent carbon quantum dot powder
End.
Marine alga according to the present invention is one or more in Enteromorpha, sea-tangle, undaria pinnitafida and seaweed, but it is several to be not limited to this
Kind, water is preferably pure water.
Enteromorpha according to the present invention is one or more in bar Enteromorpha, edge pipe Enteromorpha, shovel Enteromorpha and flat Enteromorpha.
The present invention relates to the step of (1) in by mass ratio (1.5-2.5)：1 marine alga and water are put into together tissue mashing machine
Middle crushing 3-10 minutes, it is 2 to obtain the preferred marine alga of seaweed slurry and water quality ratio：1, grinding time is 5 minutes.
The present invention relates to the step of (2) in sealing system be closed reactor, the seaweed slurry in step (2) accounts for closed anti-
1/3rd of kettle product are answered, high temperature hydro-thermal reaction and carbonisation are specially：Closed reactor is heated up in heater
To 180-200 DEG C, and maintain 60-90 minutes, be ensure fluorescent carbon quantum dot yield and quality, the temperature fluctuation of heater
Control is at ± 5 DEG C.
Heater according to the present invention is the one kind in microwave reactor, baking oven or Muffle furnace.
The present invention relates to the step of (3) in last handling process be centrifuged, filter, dialyse in one kind, but be not limited to
This is several.
Centrifugation of the present invention is specially：Carbonized product in step (2) is put in centrifuge, in rotating speed 5000-
Centrifugal treating 10-20 minute under conditions of 12000 revs/min, centrifuged supernatant is fluorescent carbon quantum dot solution, and preferred rotating speed is
10000 revs/min, centrifugation time is 15 minutes.
It is of the present invention filtration detailed process be：By carbonized product in step (2) by 0.10-0.45 μm of filter membrane
Reason, filter liquor is fluorescent carbon quantum dot solution, and preferably filter sizes are 0.22 μm.
It is of the present invention filtration detailed process be：By carbonized product in step (2) by the saturating of molecular cut off 500Da
Analysis film process removes small molecular weight impurity, and bag filter trapped inside solution is carbon quantum dot solution.
It is multiple that fluorescent carbon quantum dot with marine alga as carbon source according to the present invention is used for biomarker, Analysis of Heavy Metal, electrode
The all directions such as condensation material preparation, antibody coupling and composite photo-catalyst preparation.
Compared with prior art, the present invention has advantages below：(1) effectively by cheap marine alga, particularly Enteromorpha turn
The carbon quantum dot of the nano-scale with fluorescent characteristic is turned to, the high added value of Enteromorpha and harmless conversion is realized；(2) preparation method
Process is simple, strong operability, low cost, preparation that large-scale carbon quantum dot can be realized, without using other chemicals,
Environmental friendliness；(3) fluorescent carbon quantum dot for preparing is used for biomarker, Analysis of Heavy Metal, antibody labeling and cell dyeing；(4)
Product yield is high in preparation process；(5) comminution process is conducive to the composition in marine alga to disperse to improve follow-up in aqueous
The efficiency of hydro-thermal reaction and carbonisation.
Description of the drawings：
Fig. 1 is the transmission electron microscope phenogram of the fluorescent carbon quantum dot of embodiment according to the present invention 1；
Fig. 2 is the uv-visible absorption spectrum figure of the fluorescent carbon quantum dot of embodiment according to the present invention 1；
Fig. 3 is the fluorescent emission spectrogram of the fluorescent carbon quantum dot of embodiment according to the present invention 1；
Fig. 4 is the impact of the fluorescent carbon quantum dot cell growth of embodiment according to the present invention 1 breeding；
Fig. 5 be the concentration 500mg/L fluorescent carbon quantum dot of embodiment according to the present invention 1 and cell co-culture 5 hours it
Confocal microscopy imaging results afterwards.
Specific embodiment
With reference to specific embodiment, the present invention will be further described：
Embodiment 1：
(1) Enteromorpha is washed with running water, removes the impurity such as surface silt, be then rinsed with pure water；
(2) the Enteromorpha raw material (about 10 grams of dry weight) after 200 grams of steps (1) are processed is weighed, according to Enteromorpha and the quality of pure water
Than for 2:1 ratio mixes Enteromorpha and pure water, is put into the material container of tissue mashing machine, crushes 5 minutes, obtains Enteromorpha slurry
Material；
(3) weigh the Enteromorpha slurry that 10 grams of steps (2) obtain, be put into 30mL polytetrafluoros hydrothermal reaction kettle and adding a cover be put into it is outer
Canning tube carries out closed；
(4) reactor that will be equipped with Enteromorpha slurry is put into microwave reactor, and it is 200 DEG C to arrange temperature, and pressure is 20atm,
Reaction 60 minutes, then stops heating；
(5) temperature of question response kettle is reduced to less than 40 DEG C, and the lid of reactor is opened, and product is that bronzing is molten
Liquid, the insoluble matter containing a little black；
(6) reaction product mixture that step (5) is obtained is collected in centrifuge tube, setting speed 10000rpm, is centrifuged
20 minutes, centrifuge tube supernatant liquor is transferred in clean test tube, insoluble matter precipitation is directly abandoned；
(7) the fluorescent carbon quantum dot solution that step (6) is obtained is carried out into freeze-drying process, obtains fluorescent carbon quantum dot powder
End, it is 4.7% to calculate fluorescence quantum yield, and product yield is 9.6%.
Carbon quantum dot powder prepared by embodiment 1 is tested, as a result as Figure 1-5, from transmission electron microscope phenogram
It can be seen that the fluorescent carbon quantum dot particle for preparing is uniformly dispersed, and particle size distribution range relative narrower, substantially in 2-
In the range of 6nm, average particulate diameter is 3.5nm.Can be seen that fluorescent carbon quantum dot without obvious from uv-visible absorption spectrum figure
Absworption peak.Can draw under conditions of being irradiated to carbon quantum dot with the exciting light of 360nm from fluorescent emission spectrogram, it is glimmering
Light carbon quantum dot has very strong fluorescent emission in 400-600nm intervals, and emission peak is near 440nm.From with cell co-culture 5
Confocal microscopy imaging results after hour can draw：Growth of the Enteromorpha carbon quantum dot of concentration 500ug/mL to cell
Propagation has no significant effect, and can be enriched in cytoplasmic position carries out fluorescence labeling to it.
Embodiment 2：
(1) Enteromorpha is washed with running water, removes the impurity such as surface silt, be then rinsed with pure water；
(2) the Enteromorpha raw material (about 10 grams of dry weight) after 200 grams of steps (1) are processed is weighed, according to Enteromorpha and the quality of pure water
Than for 2:1 ratio mixes Enteromorpha and pure water, is put into and smashs cup to pieces, is crushed 5 minutes using tissue mashing machine, obtains Enteromorpha slurry
Material；
(3) the Enteromorpha slurry that 30 grams of steps (2) obtain is weighed, 100mL polytetrafluoros hydrothermal reaction kettle is put into and is added a cover and be put into
Kalamein sleeve pipe carries out closed；
(4) reactor that will be equipped with Enteromorpha slurry is put into baking oven, and it is 180 DEG C to arrange temperature, is reacted 90 minutes, course of reaction
Middle temperature fluctuation is controlled at ± 5 DEG C, then stops heating；
(5) temperature of question response kettle is reduced to less than 40 DEG C, and the lid of reactor is opened, and product is that bronzing is molten
Liquid, the insoluble matter containing a little black；
(6) reaction product mixture for obtaining step (5) is carried out at filtration with the Suction filtration device equipped with 0.22 μm of filter membrane
Reason, collects the filter liquor of clarification, and in being transferred to clean test tube, the insoluble matter that filter membrane is collected directly is abandoned；
(7) the fluorescent carbon quantum dot solution that step (6) is obtained is carried out into freeze-drying process, obtains fluorescent carbon quantum dot powder
End.It is 4.9% to calculate fluorescence quantum yield, and product yield is 7.8%.
Embodiment 3：
(1) Enteromorpha is washed with running water, removes the impurity such as surface silt, be then rinsed with pure water；
(2) the Enteromorpha raw material (about 10 grams of dry weight) after 200 grams of steps (1) are processed is weighed, according to Enteromorpha and the quality of pure water
Than for 2:1 ratio mixes Enteromorpha and pure water, is put into and smashs cup to pieces, is crushed 5 minutes using tissue mashing machine, obtains Enteromorpha slurry
Material；
(3) the Enteromorpha slurry that 30 grams of steps (2) obtain is weighed, 100mL polytetrafluoros hydrothermal reaction kettle is put into and is added a cover and be put into
Kalamein sleeve pipe carries out closed；
(4) reactor that will be equipped with Enteromorpha slurry is put into Muffle furnace, and it is 200 DEG C to arrange temperature, is reacted 60 minutes, is reacted
Temperature fluctuation in journey is controlled at ± 5 DEG C, then stops heating；
(5) temperature of question response kettle is reduced to less than 40 DEG C, and the lid of reactor is opened, and product is that bronzing is molten
Liquid, the insoluble matter containing a little black；
(6) reaction product mixture for obtaining step (5) is collected and is dispensed in centrifuge tube, setting speed 10000rpm,
Centrifugation 20 minutes, centrifuge tube supernatant liquor is transferred in clean test tube, and insoluble matter precipitation is directly abandoned；
(7) the fluorescent carbon quantum dot solution that step (6) is obtained is carried out into freeze-drying process, obtains fluorescent carbon quantum dot powder
End.It is 3.8% to calculate fluorescence quantum yield, and product yield is 8.9%.
Embodiment 4：
The present embodiment 4 and embodiment 1 are undaria pinnitafida except the raw material selected in step (1), undaria pinnitafida and pure water in step (2)
Mass ratio be 2.5:1, rotating speed 5000rpm in step (6), are centrifuged 10 minutes, and other step all sames prepare fluorescence carbon amounts
Son point powder.
Embodiment 5：
The present embodiment 5 and embodiment 2 remove other step all sames, except the raw material selected in step (1) is seaweed, step
(2) mass ratio of seaweed and pure water is 1.5 in:1,0.45 μm of filter membrane is adopted in step (6) Suction filtration device, other steps are homogeneous
Together, fluorescent carbon quantum dot powder is prepared.
Test shows, the fluorescence carbon amounts that fluorescent carbon quantum dot powder property prepared by embodiment 2-5 is prepared with embodiment 1
Son point powder property is close to.
Comparative example 1：
It is 150 DEG C that this comparative example 1 removes step (4) reaction temperature with embodiment 1, when the reaction time is 30min, other steps
All same, prepares fluorescent carbon quantum dot powder, and it is 1.2% to calculate fluorescence quantum yield, and product yield is 2.1%.
Claims (10)
1. a kind of preparation method of the fluorescent carbon quantum dot with marine alga as carbon source, it is characterised in that specifically include following steps：
(1) marine alga raw material is cleaned to go the removal of impurity, then together crush marine alga and water, obtain seaweed slurry；
(2) appropriate seaweed slurry is put in sealing system carries out high temperature hydro-thermal reaction and makes marine alga slurry be carbonized；
(3) carbonized product in step (2) is post-processed to remove insoluble matter and large-sized component, is obtained fluorescence carbon amounts
Son point solution；
(4) the fluorescent carbon quantum dot solution in step (3) is carried out into freeze-drying process, obtains fluorescent carbon quantum dot powder.
2. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 1, it is characterised in that marine alga
For one or more in Enteromorpha, sea-tangle, undaria pinnitafida and seaweed.
3. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 2, it is characterised in that Enteromorpha
For one or more in bar Enteromorpha, edge pipe Enteromorpha, shovel Enteromorpha and flat Enteromorpha.
4. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 3, it is characterised in that step
(1) by mass ratio (1.5-2.5) in：1 marine alga and water are put into together crushing 3-10 minutes in tissue mashing machine, obtain marine alga slurry
Material.
5. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 4, it is characterised in that marine alga
It is 2 with water quality ratio：1, grinding time is 5 minutes.
6. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 4 or 5, it is characterised in that
Sealing system is closed reactor in step (2), and the seaweed slurry in step (2) accounts for 1/3rd of closed reactor volume,
High temperature hydro-thermal reaction and carbonisation are specially：Closed reactor is warming up to into 180-200 DEG C in heater, and is maintained
60-90 minutes, the temperature fluctuation of heater is controlled at ± 5 DEG C.
7. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 6, it is characterised in that heating
Device is the one kind in microwave reactor, baking oven or Muffle furnace, and water is pure water, and the last handling process in step (3) is centrifugation, mistake
One kind in filter, dialysis, but it is several to be not limited to this.
8. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 7, it is characterised in that centrifugation
Specially：Carbonized product in step (2) is put in centrifuge, under conditions of 5000-12000 rev/min of rotating speed at centrifugation
Reason 10-20 minutes, centrifuged supernatant is fluorescent carbon quantum dot solution, and described filtration detailed process is：By carbonization in step (2)
Product is processed by 0.10-0.45 μm of filter membrane, and filter liquor is fluorescent carbon quantum dot solution, is filtered detailed process and is：By step (2)
Middle carbonized product removes small molecular weight impurity by the dialysis film process of molecular cut off 500Da, and bag filter trapped inside solution is
Carbon quantum dot solution.
9. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 8, it is characterised in that centrifugation
Rotating speed is 10000 revs/min, and centrifugation time is 15 minutes, and filter process filter sizes are 0.22 μm.
10. the preparation method of the fluorescent carbon quantum dot with marine alga as carbon source according to claim 1-9 Arbitrary Term, its feature
It is that fluorescent carbon quantum dot is used for biomarker, Analysis of Heavy Metal, electrode composite material preparation, antibody coupling and complex light
Catalyst preparation.
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