CN104560035B - Carbon quantum dot fluorescence labeling material with orange peels used as carbon source as well as preparation method and application of carbon quantum dot fluorescence labeling material - Google Patents
Carbon quantum dot fluorescence labeling material with orange peels used as carbon source as well as preparation method and application of carbon quantum dot fluorescence labeling material Download PDFInfo
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Abstract
The invention relates to a carbon quantum dot fluorescence labeling material with orange peels used as a carbon source as well as a preparation method and an application of the carbon quantum dot fluorescence labeling material. The orange peels are used as a raw material, and a simple method for one-step synthesis of a high-fluorescence-quantum-efficiency carbon quantum dot is established. The method comprises steps as follows: the orange peels are mixed with water; then a mixed liquid is mixed with concentrated sulfuric acid; the pH is adjusted by the aid of NaOH, and undissolved residues are removed in a centrifugal manner; then dichloromethane is added to a centrifuged supernate, and a non-fluorescence fat-soluble substance is removed in the centrifugal manner; finally, the centrifuged supernate passes through a film, and the fluorescence labeling material is prepared. According to the method, high-temperature and high-pressure reactions are not required to be performed by equipment such as plasma emitters, hydrothermal reaction kettles, muffle furnaces and the like during synthesis, reaction conditions are mild, operations are simple and convenient, the cost is very low, the method can be implemented in ordinary laboratories, the synthesized fluorescence labeling material lays a foundation for the further application to work such as bio-labeling, heavy metal analysis, antibody labeling, cell staining and the like.
Description
Technical field
The present invention relates to field of material synthesis technology, more particularly, to a kind of carbon quantum dot fluorescence mark with peel of Citrus reticulata Blanco as carbon source
Note material and its preparation method and application.
Background technology
Fluorescent carbon point as the newcomer in carbon nanometer family, be a kind of scattered, be smaller in size than the spherical of 10nm
Nano-particle, has the excellent fluorescence property such as excitation wavelength and launch wavelength is tunable, fluorescent stabilization and unglazed scintillation.
With respect to traditional semiconductor-quantum-point, rare-earth fluorescence labeling material and organic fluorescent dye dyestuff, there is particle diameter and molecular weight
Little, bio-toxicity is low, good biocompatibility the advantages of, and light emitting region is adjustable, good light stability, no optical flare, be easy to work(
The advantages such as energyization, have very big application prospect in fields such as cell imaging, biochemical analysises, develop very fast in recent years.
The carbon source of carbon quantum dot is from initial CNT, graphite, white carbon black, wax candle ash, natural gas flue dust, citrate etc.
Containing carbon organic chemical reagent, gradually arrive activated carbon, saccharide, starch, dimension c, carbon fiber, then ash wood, peanut skin, west till now
The waste product such as melon skin and coffee groundss can simply synthesize the carbon quantum dot of various visible rays.
Cn 104030269a discloses a kind of method preparing red carbon quantum dot, the method using bagasse as carbon source,
By the mixed solution heating of bagasse and phosphoric acid and sulphuric acid, obtain red carbon quantum dot after filter washing.The method is using a kind of
Biological waste resource, is effectively realized resource reutilization.Cn 103911149a discloses a kind of carbon nitrogen quantity based on ionic liquid
The preparation method of son point and its application in drug detection as fluorescence labeling material.The method is by ionic liquid and distillation
After water mix homogeneously, put into microwave oven and Muffle furnace carries out reacting by heating, obtain water miscible carbon nitrogen quantum dot, then to be prepared into
The quantum dot arriving, as fluorescence labeling material, measures the content of medicine.The method synthesis is simple, however it is necessary that Muffle furnace, microwave
The equipment such as stove carry out high-temperature high-voltage reaction, and are difficult to the ionic liquid prepared as auxiliary agent.
Fluorescent carbon quantum dot, as novel fluorescence carbon nanomaterial, not only has excellent optical property and small size property
The advantages of, or environmentally friendly new material is although also achieve certain progress in terms of preparation, but how to use easy, warp
The method of Ji prepares the more excellent carbon point of performance also to be needed further to study.
Content of the invention
The invention provides a kind of carbon quantum dot fluorescence labeling material with peel of Citrus reticulata Blanco as carbon source and preparation method thereof and should
With, methods described in building-up process, without the equipment such as plasma emitters, hydrothermal reaction kettle or Muffle furnace carry out high temperature,
Reaction under high pressure is it is only necessary to the routine instrument device such as water-bath, you can be obtained.
In a first aspect, a kind of the invention provides preparation side of the carbon quantum dot fluorescence labeling material with peel of Citrus reticulata Blanco as carbon source
Method, comprises the following steps:
(1) peel of Citrus reticulata Blanco is mixed in proportion with water；
(2) add concentrated sulphuric acid in the mixed liquor of step (1), stir, until peel of Citrus reticulata Blanco carbonization, solution changes color；
(3) adjust ph, then be centrifuged the undissolved residue of removal；
(4) dichloromethane is added the supernatant of step (3), then be centrifuged the non-blooming liposoluble substance of removal；
(5) supernatant of step (4) is crossed film, prepared fluorescence labeling material.
Described peel of Citrus reticulata Blanco is fresh peel of Citrus reticulata Blanco, for example can be selected from Pericarpium Citri tangerinae, Pericarpium Citri junoriss or Pericarpium Citri grandiss, preferably Pericarpium Citri junoriss or Fructus Citri grandiss
Skin.
Preferably, the peel of Citrus reticulata Blanco described in step (1) and water are 1:(10-40 by mass volume ratio (g:ml)) mixed,
Can be for example 1:10,1:12,1:15,1:18,1:20,1:22,1:25,1:28,1:30,1:32,1:35,1:38 or 1:40,
It is preferably 1:(15-30), more preferably 1:20.
Preferably, concentrated sulphuric acid is more than or equal to 98% sulphuric acid for mass fraction.
Preferably, the mixed liquor described in step (2) is (10-2) with concentrated sulphuric acid by volume: 1 is mixed, for example permissible
It is 10:1,5:1,10:3,5:2 or 2:1, preferably 10:3.
Preferably, the time of the described stirring of step (2) be 3-10min, can be for example 3min, 4min, 5min, 6min,
7min, 8min, 9min or 10min, preferably 4-8min, more preferably 5min.
Preferably, regulation ph to the 5.5-8.5 described in step (3), for example, can be 5.5,6,6.5,7,7.5,8 or 8.5,
It is preferably 6-8.
Preferably, the addition of the described dichloromethane of step (4) be 1-8ml, can be for example 1ml, 2ml, 3ml, 4ml,
5ml, 6ml, 7ml or 8ml, preferably 2-5ml, more preferably 3ml.
Preferably, the rotating speed of step (3) and (4) described centrifugation is 4000-8000r/min, for example, can be 4000r/
Min, 4500r/min, 5000r/min, 5500r/min, 6000r/min, 6500r/min, 7000r/min, 7500r/min or
8000r/min, preferably 4500-6000r/min, more preferably 5000r/min.
Preferably, the time of step (3) and (4) described centrifugation be 3-10min, can be for example 3min, 4min, 5min,
6min, 7min, 8min, 9min or 10min, preferably 4-8min, more preferably 5min.
Preferably, the aperture of the filter membrane described in step (5) is 0.1-1.2 μm, can be for example 0.1 μm, 0.22 μm, 0.45
μm, 0.65 μm or 1.2 μm, preferably 0.45 μm.
The invention provides a kind of preparation method of the carbon quantum dot fluorescence labeling material with peel of Citrus reticulata Blanco as carbon source, including such as
Lower step:
(1) the fresh peel of Citrus reticulata Blanco shredding is mixed by mass volume ratio 1:20g/ml with water；
(2) add concentrated sulphuric acid in the mixed liquor of step (1), mixed liquor is 10:3 with the volume ratio of concentrated sulphuric acid, stirring
5min, until peel of Citrus reticulata Blanco carbonization, solution changes color；
(3) add naoh, adjust ph to 7, be centrifuged 5min in 5000r/min, remove undissolved residue；
(4) dichloromethane of 3ml is added the supernatant of step (3), be centrifuged 5min in 5000r/min, remove unstressed configuration
Liposoluble substance；
(5) supernatant of step (4) is crossed 0.45 μm of film, prepared fluorescence labeling material.
Second aspect, present invention also offers a kind of carbon quantum dot of method preparation as described in the first aspect of the invention is glimmering
Signal material.
The third aspect, present invention also offers carbon quantum dot fluorescence labeling material as described in respect of the second aspect of the invention is being given birth to
Substance markers and the application of bio-imaging aspect.
Compared with prior art, the present invention has the advantage that
(1) present invention peel of Citrus reticulata Blanco prepares carbon quantum dot fluorescence labeling material as carbon source, and raw material is cheap and easy to get, source
Abundant, be conducive to the abundant recycling of waste resource；
(2) the preparation method equipment of the carbon quantum dot fluorescence labeling material of the present invention is simple, and building-up process is without plasma
The equipment such as body emitter, hydrothermal reaction kettle or Muffle furnace carries out high temperature, reaction under high pressure, and reaction condition is gently carried out at normal temperatures,
Response time is short, simple to operate, with low cost, small toxicity；
(3) the carbon quantum dot fluorescence labeling material of the present invention as fluorescent marker be widely used in biomarker with biological become
Picture
Brief description
Fig. 1 is the transmission electron microscope photo of the specific embodiment of the invention 1.
Fig. 2 is the carbon quantum dot fluorescence spectral characteristic figure with peel of Citrus reticulata Blanco as carbon source in the specific embodiment of the invention 1.
Specific embodiment
For further illustrating the technological means and its effect that the present invention taken, being preferable to carry out below in conjunction with the present invention
Example is further illustrating technical scheme, but the present invention is not limited in scope of embodiments.
Embodiment 1
A kind of carbon quantum dot fluorescence labeling material preparation method with Pericarpium Citri grandiss as carbon source, comprises the following steps:
(1) the fresh Pericarpium Citri grandiss 0.5g shredding is taken to add in 10ml water, stir about 5min；
(2) add dense h in step (1)2so4Solution 3ml, stir about 5min, until Pericarpium Citri grandiss carbonization, solution changes color；
(3) add naoh solution, adjust ph to 7, then remove undissolved residue in rotating speed 5000r/min centrifugation 5min；
(4) dichloromethane of 3ml is added the supernatant of step (3), then remove no in rotating speed 5000r/min centrifugation 5min
The liposoluble substance of fluorescence；
(5) supernatant of step (4) is crossed 0.45 μm of filter membrane, prepared fluorescence labeling material.
The Pericarpium Citri grandiss prepared as can be seen from Figure 1 are the carbon quantum dot fluorescence labeling material of carbon source is near-spherical, monodispersity
Good, do not assemble, particle diameter is in 50nm；The obtained carbon quantum dot fluorescence mark with Pericarpium Citri grandiss as carbon source as can be seen from Figure 2
Note material, maximum excitation wavelength 422nm, maximum emission wavelength 504nm, can be effectively as fluorescence labeling material.
Embodiment 2
A kind of carbon quantum dot fluorescence labeling material preparation method with Pericarpium Citri grandiss as carbon source, comprises the following steps:
(1) the fresh Pericarpium Citri grandiss 0.7g shredding is taken to add in 10ml water, stir about 5min；
(2) add dense h in step (1)2so4Solution 3ml, stir about 5min, until Pericarpium Citri grandiss carbonization, solution changes color；
(3) add naoh solution, adjust ph to 7.5, then remove in rotating speed 5500r/min centrifugation 5min undissolved residual
Slag；
(4) dichloromethane of 1ml is added the supernatant of step (3), then remove no in rotating speed 5500r/min centrifugation 5min
The liposoluble substance of fluorescence；
(5) supernatant of step (4) is crossed 0.1 μm of filter membrane, prepared fluorescence labeling material.
Embodiment 3
A kind of carbon quantum dot fluorescence labeling material preparation method with Pericarpium Citri grandiss as carbon source, comprises the following steps:
(1) the fresh Pericarpium Citri grandiss 0.3g shredding is taken to add in 10ml water, stir about 3min；
(2) add dense h in step (1)2so4Solution 3ml, stir about 5min, until Pericarpium Citri grandiss carbonization, solution changes color；
(3) add naoh solution, adjust ph to 7, then remove undissolved residue in rotating speed 6000r/min centrifugation 5min；
(4) dichloromethane of 5ml is added the supernatant of step (3), then remove no in rotating speed 6000r/min centrifugation 5min
The liposoluble substance of fluorescence；
(5) supernatant of step (4) is crossed 0.22 μm of filter membrane, prepared fluorescence labeling material.
Embodiment 4
A kind of carbon quantum dot fluorescence labeling material preparation method with Pericarpium Citri junoriss as carbon source, comprises the following steps:
(1) the fresh Pericarpium Citri junoriss 0.5g shredding is taken to add in 10ml water, stir about 5min；
(2) add dense h in step (1)2so4Solution 1ml, stir about 3min, until Pericarpium Citri junoriss carbonization, solution changes color；
(3) add 5m naoh solution, adjust ph to 7, then remove in rotating speed 8000r/min centrifugation 3min undissolved residual
Slag；
(4) dichloromethane of 8ml is added the supernatant of step (3), then remove no in rotating speed 8000r/min centrifugation 5min
The liposoluble substance of fluorescence；
(5) supernatant of step (4) is crossed 1.2 μm of filter membrane, prepared fluorescence labeling material.
Embodiment 5
A kind of carbon quantum dot fluorescence labeling material preparation method with Fructus Citri sinensiss as carbon source, comprises the following steps:
(1) the fresh Pericarpium Citri junoriss 0.5g shredding is taken to add in 10ml water, stir about 5min；
(2) add dense h in step (1)2so4Solution 5ml, stir about 8min, until Pericarpium Citri junoriss carbonization, solution changes color；
(3) add naoh solution, adjust ph to 7.2, then remove in rotating speed 4000r/min centrifugation 6min undissolved residual
Slag；
(4) dichloromethane of 3ml is added the supernatant of step (3), then remove no in rotating speed 4000r/min centrifugation 6min
The liposoluble substance of fluorescence；
(5) supernatant of step (4) is crossed 0.45 μm of filter membrane, prepared fluorescence labeling material.
The carbon quantum dot fluorescence labeling material that embodiment 1-4 makes and fluorescent effect are similar to Example 1.
Integrated embodiment 1-5 it will thus be seen that
Embodiment 1-5 is reproducible, shows that the marker material stability made is high.This method in building-up process,
Without complex device, reaction condition is gentle, easy and simple to handle, using cheap and simple peel of Citrus reticulata Blanco as raw material, cost is very low, typically
Laboratory all can be carried out, the fluorescence labeling material of synthesis for next step be applied to biomarker, Analysis of Heavy Metal, antibody labeling,
The work such as cell dyeing are laid a good foundation..
Applicant states, the present invention illustrates the method detailed of the present invention by above-described embodiment, but the present invention not office
It is limited to above-mentioned method detailed, that is, do not mean that the present invention has to rely on above-mentioned method detailed and could implement.Art
Technical staff is it will be clearly understood that any improvement in the present invention, the equivalence replacement to each raw material of product of the present invention and auxiliary element
Interpolation, selection of concrete mode etc., within the scope of all falling within protection scope of the present invention and disclosure.
Claims (20)
1. a kind of preparation method of the carbon quantum dot fluorescence labeling material with peel of Citrus reticulata Blanco as carbon source is it is characterised in that include as follows
Step:
(1) peel of Citrus reticulata Blanco is mixed in proportion with water；
(2) add concentrated sulphuric acid in the mixed liquor of step (1), stir up to peel of Citrus reticulata Blanco carbonization, solution changes color；
(3) adjust ph, then be centrifuged the undissolved residue of removal；
(4) dichloromethane is added the supernatant of step (3), centrifugation removes non-blooming liposoluble substance；
(5) supernatant of step (4) is crossed film, prepared fluorescence labeling material；
Peel of Citrus reticulata Blanco described in step (1) and water are 1:(10-40 by mass volume ratio g/ml) mixed；
Mixed liquor described in step (2) is (10-2) with concentrated sulphuric acid by volume: 1 is mixed；
Regulation ph to 5.5-8.5 described in step (3)；
The addition of the described dichloromethane of step (4) is 1-8ml.
2. method according to claim 1 is it is characterised in that the peel of Citrus reticulata Blanco described in step (1) presses mass volume ratio with water
G/ml is 1:(15-30) mixed.
3. method according to claim 2 is it is characterised in that the peel of Citrus reticulata Blanco described in step (1) presses mass volume ratio with water
G/ml is mixed for 1:20.
4. method according to claim 1 it is characterised in that mixed liquor described in step (2) with concentrated sulphuric acid by volume
Mixed for 10:3.
5. method according to claim 1 is it is characterised in that the time of the described stirring of step (2) is 3-10min.
6. method according to claim 5 is it is characterised in that the time of the described stirring of step (2) is 4-8min.
7. method according to claim 6 is it is characterised in that the time of the described stirring of step (2) is 5min.
8. method according to claim 1 is it is characterised in that regulation ph to 6-8 described in step (3).
9. method according to claim 8 is it is characterised in that regulation ph to 7 described in step (3).
10. method according to claim 1 is it is characterised in that the addition of the described dichloromethane of step (4) is 2-5ml.
11. methods according to claim 10 are it is characterised in that the addition of the described dichloromethane of step (4) is 3ml.
12. methods according to claim 1 are it is characterised in that the rotating speed of step (3) and (4) described centrifugation is 4000-
8000r/min.
13. methods according to claim 12 are it is characterised in that the rotating speed of step (3) and (4) described centrifugation is 4500-
6000r/min.
14. methods according to claim 13 are it is characterised in that the rotating speed of step (3) and (4) described centrifugation is 5000r/
min.
15. methods according to claim 1 are it is characterised in that the time of step (3) and (4) described centrifugation is 3-
10min.
16. methods according to claim 15 are it is characterised in that the time of step (3) and (4) described centrifugation is 4-
8min.
17. methods according to claim 16 are it is characterised in that the time of step (3) and (4) described centrifugation is 5min.
18. methods according to claim 1 are it is characterised in that the aperture of filter membrane described in step (5) is 0.1-1.2 μm.
19. methods according to claim 18 are it is characterised in that the aperture of filter membrane described in step (5) is 0.45 μm.
20. methods according to claim 1 are it is characterised in that comprise the steps:
(1) the fresh peel of Citrus reticulata Blanco shredding is mixed by mass volume ratio 1:20g/ml with water；
(2) add concentrated sulphuric acid in the mixed liquor of step (1), mixed liquor is 10:3 with the volume ratio of concentrated sulphuric acid, stirs 5min, directly
To peel of Citrus reticulata Blanco carbonization, solution changes color；
(3) add naoh, adjust ph to 7, be centrifuged 5min in 5000r/min, remove undissolved residue；
(4) dichloromethane of 3ml is added the supernatant of step (3), be centrifuged 5min in 5000r/min, remove non-blooming fat
Soluble substance；
(5) supernatant of step (4) is crossed 0.45 μm of film, prepared fluorescence labeling material.
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| CN105954247B (en) * | 2016-05-04 | 2019-03-12 | 东南大学 | A kind of mercury ion is luminous to indicate material and its preparation method and application |
| CN106047343B (en) * | 2016-06-27 | 2018-08-07 | 青岛大学 | The method for using bulk kelp to prepare fluorescent carbon nano dot for carbon source one-step method hydrothermal carbonization |
| CN106833630B (en) * | 2017-01-22 | 2019-11-05 | 桂林碳谷科技有限公司 | A kind of pH response type blue-fluorescence carbon quantum dot and its preparation method and application |
| CN107446578A (en) * | 2017-07-20 | 2017-12-08 | 中国科学院理化技术研究所 | A kind of nitrogen-doped carbon quanta point material and its preparation method and application |
| CN107601457A (en) * | 2017-11-07 | 2018-01-19 | 辽宁工业大学 | A kind of method that carbon quantum dot is prepared using bagasse as utilization of carbon source chemical oxidization method |
| CN109486486B (en) * | 2018-12-21 | 2021-12-17 | 达州职业技术学院 | Preparation method of fluorescent carbon quantum dots based on microwave method |
| CN110702754B (en) * | 2019-11-08 | 2021-05-04 | 广西师范大学 | Method for measuring human chorionic gonadotropin |
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| CN103834395A (en) * | 2012-11-23 | 2014-06-04 | 天津工业大学 | Method for preparing luminescent carbon dots from glycerin |
| CN104130777B (en) * | 2013-05-03 | 2016-06-22 | 首都医科大学 | A kind of fluorescent carbon point and synthetic method thereof and its application in cell marking |
| CN103588189B (en) * | 2013-07-04 | 2015-07-15 | 上海交通大学 | Egg-based method for realizing hydrothermal synthesis of carbon quantum dots |
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