CN104560035A - 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, particularly relating to a kind of take peel of Citrus reticulata Blanco as carbon quantum dot fluorescence labeling material of carbon source and its preparation method and application.
Background technology
Fluorescent carbon point, as the newcomer in carbon nanometer family, is a kind of dispersion, size is less than the spherical nano particle of the class of 10nm, has excitation wavelength and emission wavelength is tunable, fluorescent stabilization and the excellent fluorescence property such as unglazed scintillation.Relative to traditional semiconductor-quantum-point, rare-earth fluorescence labeling material and organic fluorescent dye dyestuff, there is particle diameter and the advantage such as molecular weight is little, bio-toxicity is low, good biocompatibility, and light emitting region is adjustable, good light stability, without optical flare, be easy to the advantages such as functionalization, have very large application prospect in the field such as cell imaging, biochemical analysis, development in recent years is very fast.
The carbon source of carbon quantum dot contains carbon organic chemical reagent from initial carbon nanotube, graphite, carbon black, candle ash, Sweet natural gas cigarette ash, Citrate trianion etc., arrive gac, carbohydrate, starch, Vc, carbon fiber gradually, then the waste product such as ash wood, peanut skin, Watermelon rind and coffee grounds simply can synthesize the carbon quantum dot of various visible ray till now.
CN 104030269A discloses a kind of method preparing red carbon quantum dot, and the mixing solutions of bagasse and phosphoric acid and sulfuric acid, using bagasse as carbon source, heats by the method, refilters washing and obtains red carbon quantum dot.The method uses a kind of biological waste resource, effectively realizes resource reutilization.CN 103911149A disclose a kind of carbon nitrogen quantum dot based on ionic liquid preparation method and as the application of fluorescence labeling material in drug testing.The method is after ionic liquid and distilled water being mixed, and puts into microwave oven and retort furnace carries out reacting by heating, obtains water miscible carbon nitrogen quantum dot, then using the quantum dot prepared as fluorescence labeling material, measures the content of medicine.The method synthesis is simple, but needs the equipment such as retort furnace, microwave oven to carry out high-temperature high-voltage reaction, and the ionic liquid being difficult to prepare is as auxiliary agent.
Fluorescent carbon quantum dot is as novel fluorescence carbon nanomaterial, not only there is the advantages such as excellent optical property and small size property, or environmentally friendly novel material, although also achieve certain progress in preparation, how also to need further research with the carbon point that easy, economic method prepares performance more excellent.
Summary of the invention
The invention provides a kind of take peel of Citrus reticulata Blanco as carbon quantum dot fluorescence labeling material of carbon source and its preparation method and application, described method is in building-up process, high temperature, reaction under high pressure is carried out without the need to equipments such as plasma emitters, hydrothermal reaction kettle or retort furnaces, only need the routine instrument devices such as water-bath, can obtain.
First aspect, the invention provides a kind of take peel of Citrus reticulata Blanco as the preparation method of the carbon quantum dot fluorescence labeling material of carbon source, comprises the following steps:
(1) peel of Citrus reticulata Blanco and water are mixed in proportion;
(2) in the mixed solution of step (1), add the vitriol oil, stir, until peel of Citrus reticulata Blanco carbonization, solution changes color;
(3) pH is regulated, the more undissolved residue of centrifugal segregation;
(4) methylene dichloride is added the supernatant liquor of step (3), the more non-blooming lipid-soluble substance of centrifugal segregation;
(5) supernatant liquor of step (4) is crossed film, obtained fluorescence labeling material.
Described peel of Citrus reticulata Blanco is fresh peel of Citrus reticulata Blanco, such as, can be selected from orange peel, orange peel or shaddock ped, is preferably orange peel or shaddock ped.
Preferably, peel of Citrus reticulata Blanco described in step (1) and water are 1:(10-40 by mass volume ratio (g:mL)) mix, can be such as 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, be preferably 1:(15-30), more preferably 1:20.
Preferably, the vitriol oil is the sulfuric acid that massfraction is more than or equal to 98%.
Preferably, the mixed solution described in step (2) and the vitriol oil are (10-2) by volume: 1 mixes, such as, can be 10:1,5:1,10:3,5:2 or 2:1, are preferably 10:3.
Preferably, the time of step (2) described stirring is 3-10min, such as, can be 3min, 4min, 5min, 6min, 7min, 8min, 9min or 10min, is preferably 4-8min, more preferably 5min.
Preferably, adjustment pH to the 5.5-8.5 described in step (3) can be such as 5.5,6,6.5,7,7.5,8 or 8.5, is preferably 6-8.
Preferably, the add-on of step (4) described methylene dichloride is 1-8mL, such as, can be 1mL, 2mL, 3mL, 4mL, 5mL, 6mL, 7mL or 8mL, is preferably 2-5mL, more preferably 3mL.
Preferably, step (3) and (4) described centrifugal rotating speed are 4000-8000r/min, can be such as 4000r/min, 4500r/min, 5000r/min, 5500r/min, 6000r/min, 6500r/min, 7000r/min, 7500r/min or 8000r/min, be preferably 4500-6000r/min, more preferably 5000r/min.
Preferably, step (3) and (4) described centrifugal time are 3-10min, such as, can be 3min, 4min, 5min, 6min, 7min, 8min, 9min or 10min, are preferably 4-8min, more preferably 5min.
Preferably, the aperture of the filter membrane described in step (5) is 0.1-1.2 μm, such as, can be 0.1 μm, 0.22 μm, 0.45 μm, 0.65 μm or 1.2 μm, is preferably 0.45 μm.
The invention provides a kind of take peel of Citrus reticulata Blanco as the preparation method of the carbon quantum dot fluorescence labeling material of carbon source, comprises the steps:
(1) the fresh peel of Citrus reticulata Blanco shredded is mixed by mass volume ratio 1:20g/mL with water;
(2) in the mixed solution of step (1), add the vitriol oil, the volume ratio of mixed solution and the vitriol oil is 10:3, stirs 5min, until peel of Citrus reticulata Blanco carbonization, and solution changes color;
(3) add NaOH, regulate pH to 7, at the centrifugal 5min of 5000r/min, remove undissolved residue;
(4) methylene dichloride of 3mL is added the supernatant liquor of step (3), at the centrifugal 5min of 5000r/min, remove non-blooming lipid-soluble substance;
(5) supernatant liquor of step (4) is crossed the film of 0.45 μm, obtained fluorescence labeling material.
Second aspect, present invention also offers carbon quantum dot fluorescence labeling material prepared by a kind of method as described in the first aspect of the invention.
The third aspect, present invention also offers the application of carbon quantum dot fluorescence labeling material in biomarker and bio-imaging as described in respect of the second aspect of the invention.
Compared with prior art, tool of the present invention has the following advantages:
(1) the present invention's peel of Citrus reticulata Blanco prepares carbon quantum dot fluorescence labeling material as carbon source, and cheaper starting materials is easy to get, abundance, is conducive to the abundant recycling of waste resource;
(2) preparation method's equipment of carbon quantum dot fluorescence labeling material of the present invention is simple, building-up process carries out high temperature, reaction under high pressure without the need to equipments such as plasma emitters, hydrothermal reaction kettle or retort furnaces, reaction conditions gentleness is carried out at normal temperatures, reaction times is short, simple to operate, with low cost, toxicity is little;
(3) carbon quantum dot fluorescence labeling material of the present invention is widely used in biomarker and bio-imaging as fluorescent marker
Accompanying drawing explanation
Fig. 1 is the transmission electron microscope photo of the specific embodiment of the invention 1.
Fig. 2 in the specific embodiment of the invention 1 take peel of Citrus reticulata Blanco as the carbon quantum dot fluorescence spectral characteristic figure of carbon source.
Embodiment
For further setting forth the technique means and effect thereof that the present invention takes, further illustrate technical scheme of the present invention below in conjunction with the preferred embodiments of the present invention, but the present invention is not confined in scope of embodiments.
Embodiment 1
Take shaddock ped as a carbon quantum dot fluorescence labeling material preparation method for carbon source, comprise the following steps:
(1) getting the fresh shaddock ped 0.5g shredded adds in 10ml water, stir about 5min;
(2) in step (1), dense H is added
2sO
4solution 3mL, stir about 5min, until shaddock ped carbonization, solution changes color;
(3) add NaOH solution, regulate pH to 7, then remove undissolved residue at the centrifugal 5min of rotating speed 5000r/min;
(4) methylene dichloride of 3mL is added the supernatant liquor of step (3), then remove non-blooming lipid-soluble substance at the centrifugal 5min of rotating speed 5000r/min;
(5) supernatant liquor of step (4) is crossed the filter membrane of 0.45 μm, obtained fluorescence labeling material.
The shaddock ped prepared as can be seen from Figure 1 is the carbon quantum dot fluorescence labeling material of carbon source is near-spherical, and monodispersity is good, and do not assemble, particle diameter is at about 50nm; Obtained as can be seen from Figure 2 take shaddock ped as the carbon quantum dot fluorescence labeling material of carbon source, and maximum excitation wavelength 422nm, maximum emission wavelength 504nm, can be effective as fluorescence labeling material.
Embodiment 2
Take shaddock ped as a carbon quantum dot fluorescence labeling material preparation method for carbon source, comprise the following steps:
(1) getting the fresh shaddock ped 0.7g shredded adds in 10ml water, stir about 5min;
(2) in step (1), dense H is added
2sO
4solution 3mL, stir about 5min, until shaddock ped carbonization, solution changes color;
(3) add NaOH solution, regulate pH to 7.5, then remove undissolved residue at the centrifugal 5min of rotating speed 5500r/min;
(4) methylene dichloride of 1mL is added the supernatant liquor of step (3), then remove non-blooming lipid-soluble substance at the centrifugal 5min of rotating speed 5500r/min;
(5) supernatant liquor of step (4) is crossed the filter membrane of 0.1 μm, obtained fluorescence labeling material.
Embodiment 3
Take shaddock ped as a carbon quantum dot fluorescence labeling material preparation method for carbon source, comprise the following steps:
(1) getting the fresh shaddock ped 0.3g shredded adds in 10ml water, stir about 3min;
(2) in step (1), dense H is added
2sO
4solution 3mL, stir about 5min, until shaddock ped carbonization, solution changes color;
(3) add NaOH solution, regulate pH to 7, then remove undissolved residue at the centrifugal 5min of rotating speed 6000r/min;
(4) methylene dichloride of 5mL is added the supernatant liquor of step (3), then remove non-blooming lipid-soluble substance at the centrifugal 5min of rotating speed 6000r/min;
(5) supernatant liquor of step (4) is crossed the filter membrane of 0.22 μm, obtained fluorescence labeling material.
Embodiment 4
Take orange peel as a carbon quantum dot fluorescence labeling material preparation method for carbon source, comprise the following steps:
(1) getting the fresh orange peel 0.5g shredded adds in 10ml water, stir about 5min;
(2) in step (1), dense H is added
2sO
4solution 1mL, stir about 3min, until orange peel carbonization, solution changes color;
(3) add 5M NaOH solution, regulate pH to 7, then remove undissolved residue at the centrifugal 3min of rotating speed 8000r/min;
(4) methylene dichloride of 8mL is added the supernatant liquor of step (3), then remove non-blooming lipid-soluble substance at the centrifugal 5min of rotating speed 8000r/min;
(5) supernatant liquor of step (4) is crossed the filter membrane of 1.2 μm, obtained fluorescence labeling material.
Embodiment 5
Take sweet orange as a carbon quantum dot fluorescence labeling material preparation method for carbon source, comprise the following steps:
(1) getting the fresh orange peel 0.5g shredded adds in 10ml water, stir about 5min;
(2) in step (1), dense H is added
2sO
4solution 5mL, stir about 8min, until orange peel carbonization, solution changes color;
(3) add NaOH solution, regulate pH to 7.2, then remove undissolved residue at the centrifugal 6min of rotating speed 4000r/min;
(4) methylene dichloride of 3mL is added the supernatant liquor of step (3), then remove non-blooming lipid-soluble substance at the centrifugal 6min of rotating speed 4000r/min;
(5) supernatant liquor of step (4) is crossed the filter membrane of 0.45 μm, obtained fluorescence labeling material.
Carbon quantum dot fluorescence labeling material and the fluorescent effect of embodiment 1-4 making are similar to Example 1.
Integrated embodiment 1-5, can find out:
Embodiment 1-5 is reproducible, shows that the marker material stability of making is high.Present method is in building-up process, without the need to complex apparatus, reaction conditions is gentle, easy and simple to handle, adopt the peel of Citrus reticulata Blanco of cheap and simple as raw material, cost is very low, and common laboratory all can be carried out, and the fluorescence labeling material of synthesis is applied to the work such as biomarker, Analysis of Heavy Metal, antibody labeling, cell dyeing and lays a good foundation for next step.。
Applicant states, the present invention illustrates method detailed of the present invention by above-described embodiment, but the present invention is not limited to above-mentioned method detailed, does not namely mean that the present invention must rely on above-mentioned method detailed and could implement.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of ancillary component, the concrete way choice etc. of each raw material of product of the present invention, all drops within protection scope of the present invention and open scope.
Claims (10)
1. be a preparation method for the carbon quantum dot fluorescence labeling material of carbon source with peel of Citrus reticulata Blanco, it is characterized in that, comprise the steps:
(1) peel of Citrus reticulata Blanco and water are mixed in proportion;
(2) in the mixed solution of step (1), add the vitriol oil, stir until peel of Citrus reticulata Blanco carbonization, solution changes color;
(3) pH is regulated, the more undissolved residue of centrifugal segregation;
(4) methylene dichloride is added the supernatant liquor of step (3), the non-blooming lipid-soluble substance of centrifugal segregation;
(5) supernatant liquor of step (4) is crossed film, obtained fluorescence labeling material.
2. method according to claim 1, it is characterized in that, peel of Citrus reticulata Blanco described in step (1) and water are 1:(10-40 by mass volume ratio g/mL) mix, be preferably 1:(15-30), more preferably 1:20.
3. method according to claim 1 and 2, is characterized in that, the mixed solution described in step (2) and the vitriol oil are (10-2) by volume: 1 mixes, and are preferably 10:3;
Preferably, the time of step (2) described stirring is 3-10min, is preferably 4-8min, more preferably 5min.
4. the method according to any one of claim 1-3, is characterized in that, adjustment pH to the 5.5-8.5 described in step (3), is preferably 6-8; More preferably 7.
5. the method according to any one of claim 1-4, is characterized in that, the add-on of step (4) described methylene dichloride is 1-8mL, is preferably 2-5mL, more preferably 3mL.
6. the method according to any one of claim 1-5, is characterized in that, step (3) and (4) described centrifugal rotating speed are 4000-8000r/min, is preferably 4500-6000r/min, more preferably 5000r/min;
Preferably, step (3) and (4) described centrifugal time are 3-10min, are preferably 4-8min, more preferably 5min.
7. the method according to any one of claim 1-6, is characterized in that, the aperture of the filter membrane described in step (5) is 0.1-1.2 μm, is preferably 0.45 μm.
8. the method according to any one of claim 1-7, is characterized in that, comprises the steps:
(1) the fresh peel of Citrus reticulata Blanco shredded is mixed by mass volume ratio 1:20g/mL with water;
(2) in the mixed solution of step (1), add the vitriol oil, the volume ratio of mixed solution and the vitriol oil is 10:3, stirs 5min, until peel of Citrus reticulata Blanco carbonization, and solution changes color;
(3) add NaOH, regulate pH to 7, at the centrifugal 5min of 5000r/min, remove undissolved residue;
(4) methylene dichloride of 3mL is added the supernatant liquor of step (3), at the centrifugal 5min of 5000r/min, remove non-blooming lipid-soluble substance;
(5) supernatant liquor of step (4) is crossed the film of 0.45 μm, obtained fluorescence labeling material.
9. carbon quantum dot fluorescence labeling material prepared by the method according to any one of claim 1-8.
10. the application of carbon quantum dot fluorescence labeling material according to claim 9 in biomarker and bio-imaging.
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