CN116103042A - Preparation method of fluorescent carbon quantum dot with castor seeds as carbon source - Google Patents
Preparation method of fluorescent carbon quantum dot with castor seeds as carbon source Download PDFInfo
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- CN116103042A CN116103042A CN202310062865.XA CN202310062865A CN116103042A CN 116103042 A CN116103042 A CN 116103042A CN 202310062865 A CN202310062865 A CN 202310062865A CN 116103042 A CN116103042 A CN 116103042A
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Abstract
The invention provides a preparation method of fluorescent carbon quantum dots by taking castor seeds as a carbon source, which comprises the following steps: dispersing peeled castor seed powder into deionized water, uniformly stirring, and placing into a hydrothermal kettle to perform hydrothermal reaction in a drying oven; taking out the product after the reaction is finished, filtering and centrifuging, taking out the supernatant, and concentrating by rotary evaporation; adding the concentrated aqueous solution into a liquid chromatographic column filled with silica gel powder, washing impurities with dichloromethane, extracting carbon quantum dots of castor seeds with deionized water, and freeze-drying to obtain powder. The surface of the carbon quantum dot prepared by the method has no adhesive organic impurities, and the lattice stripes are clearly visible. In addition, the preparation method is low in cost and simple in implementation, and the prepared carbon quantum dot is good in light intensity and dispersibility, can stably emit light in different pH environments, and is expected to be widely applied to chemical detection and biological imaging.
Description
Technical Field
The invention relates to a preparation method of carbon quantum dots, in particular to a preparation method of fluorescent carbon quantum dots with castor seeds as carbon sources.
Background
The nano fluorescent material has the advantages of high fluorescence efficiency, stable performance, strong adjustability and the like, and has great application prospect in the aspects of photoelectric devices and sensing detection. Whereas conventional inorganic semiconductor quantum dots (e.g. CdTe, cdSe) and inorganic perovskite quantum dots (e.g. CsPbI) 3 、CsPbBr 3 ) Although excellent in luminescence performance, heavy metals contained in the light-emitting material have great negative effects on organisms and environment, and the common preparation is complex and the raw materials are expensive. In recent years, a plurality of high-performance and easily-controlled carbon quantum dots are competitively reported by researchers at home and abroad. As a novel fluorescent nano material, the carbon quantum dot has low toxicity or no toxicity, good biocompatibility and water solubility, and the preparation raw materials are cheap and easy to obtain. Has very wide application space in the aspects of light emitting devices, sensing detection, biomedicine and the like.
The selection of the carbon source is important to the in-situ synthesis of the carbon quantum dot, and the effect of the carbon source is particularly obvious in the research of heteroatom doping and surface group regulation. Biomass carbon sources are abundant in nature and often contain elements such as nitrogen, phosphorus and the like, so that the biomass carbon sources have great potential for preparing excellent carbon points. However, the carbon dots prepared from the biological carbon source are often not pure enough, and most of the fluorescence does not have pH stability, in other words, the preparation of clean carbon dots with stable luminescence in different pH environments by using a cheap and easily available biological carbon source has great significance.
Disclosure of Invention
The invention aims to provide a preparation method of fluorescent carbon quantum dots with castor seeds as carbon sources, so as to obtain carbon quantum dots with stable luminescence in different pH environments.
The invention is realized in the following way: a preparation method of fluorescent carbon quantum dots by taking castor seeds as carbon sources comprises the following steps:
a. dispersing peeled castor seed powder into deionized water, uniformly stirring, and placing into a hydrothermal kettle to perform hydrothermal reaction in a drying oven;
b. taking out the product after the reaction is finished, filtering and centrifuging, taking out the supernatant, and concentrating by rotary evaporation;
c. adding the concentrated aqueous solution into a chromatographic column filled with silica gel powder, washing impurities with dichloromethane, extracting carbon quantum dots of castor seeds with deionized water, and freeze-drying to obtain powder.
Preferably, in the step a, the dosage ratio of the castor seed powder to the deionized water is 2-5 g:50ml.
Preferably, the peeled castor seed meal of step a is prepared by: and soaking the castor seeds in deionized water for 5-10 hours, peeling, grinding, sieving, and drying at 60-80 ℃ for 15-30 minutes.
Preferably, the sieving aperture of the ground castor seeds is 100-200 meshes.
Preferably, the hydrothermal kettle in the step a takes polytetrafluoroethylene as an inner container.
Preferably, in the step a, the temperature of the hydrothermal reaction is 150-180 ℃ and the reaction time is 4-8 hours.
Preferably, in the step b, the aperture of the filter head is 0.22 mu m or 0.45 mu m, the rotation speed of the centrifugal machine is 6000-8000 rpm, the centrifugation time is 10-15 min, the rotary evaporation temperature of the supernatant fluid is 65-80 ℃, and the rotary evaporation concentration is 30-40 mg/ml.
Preferably, in the step c, the particle size of the silica gel powder is 100-200 meshes, the purity of dichloromethane is more than or equal to 99.0%, the pressure in a freeze dryer cavity is 30-100pa, and the freeze drying time is 24-30 h.
The invention has the beneficial effects that:
(1) And (3) taking the natural carbon source castor seeds as a single carbon source to obtain the fluorescent carbon quantum dots with stable pH. The preparation method is low in cost and simple in implementation, and the prepared carbon quantum dots are high in purity, strong in luminescence, good in dispersibility, capable of stably emitting luminescence in different pH environments, and expected to be widely applied to chemical detection and biological imaging.
(2) Unlike the dialysis method for preparing biological carbon source carbon quantum dots, the preparation-purification system of the invention adopts a one-step hydrothermal method-column chromatography-freeze drying combination, so that the surface of the carbon quantum dots is free from adhesion of organic impurities, the lattice stripes are clear and visible, and a new thought is provided for preparing the biological carbon source carbon quantum dots.
Drawings
Fig. 1 is a transmission electron microscope image of castor seed carbon quantum dots prepared in example 2.
Fig. 2 is a high resolution transmission electron microscope image of the castor seed carbon quantum dots prepared in example 2.
FIG. 3 is an X-ray photoelectron spectrometer spectrum of the castor seed carbon quantum dot prepared in example 2.
FIG. 4 is an infrared absorption spectrum of the carbon quantum dots of castor seeds prepared in example 2.
Fig. 5 is an ultraviolet-visible absorption spectrum of the carbon quantum dots of castor seeds prepared in example 2.
Fig. 6 is a fluorescence excitation emission spectrum of the castor seed carbon quantum dot prepared in example 2.
Fig. 7 is an emission spectrum diagram of the castor seed carbon quantum dot prepared in example 2 under excitation of different excitation lights.
Fig. 8 is a fluorescence lifetime graph of the castor seed carbon quantum dots prepared in example 2.
Fig. 9 is a graph showing changes in fluorescence intensity measured at different pH values of the castor seed carbon quantum dots prepared in example 2.
Detailed Description
The invention is further illustrated by the following examples, which are given by way of illustration only and are not intended to limit the scope of the invention in any way.
The procedures and methods not described in detail in the examples below are conventional methods well known in the art, and the reagents used in the examples are all analytically or chemically pure and are either commercially available or prepared by methods well known to those of ordinary skill in the art.
EXAMPLE 1 preparation of Castor seed powder
Soaking semen Ricini in deionized water for 6 hr, taking out, peeling, grinding for 10min, sieving with 200 mesh sieve, and oven drying at 70deg.C for 20min to obtain semen Ricini powder.
Example 2 preparation of fluorescent carbon Quantum dots Using Castor seeds as carbon Source (method 1)
Dispersing 2g of the castor seed powder prepared in the example 1 into 50ml of deionized water, stirring for 10min, and putting into a hydrothermal kettle taking tetrafluoroethylene as an inner container for hydrothermal reaction, wherein the temperature of the hydrothermal reaction is 150 ℃ and the reaction time is 5h; filtering with a 0.45 mu m filter head after the reaction is finished, centrifuging at 7000rpm for 10min in a centrifuge, and concentrating the supernatant in a rotary steaming instrument at 70 ℃ to 30 mg/ml; adding the concentrated aqueous solution into column chromatography filled with 200-mesh silica gel powder, washing impurities with dichloromethane with purity of more than or equal to 99.0%, eluting with deionized water to obtain pure carbon quantum dot solution, and finally placing the pure carbon quantum dot solution into a freeze dryer, and drying at 50pa for 24h to obtain white castor seed carbon quantum dot powder.
1g of carbon quantum dot powder is dispersed into 20ml of deionized water, and stirred uniformly to obtain a carbon quantum dot aqueous solution, and the TEM particle size distribution of the carbon quantum dot aqueous solution is measured, and the measurement result is shown in figure 1, and the average particle size of the carbon quantum dots is 15nm as can be seen from figure 1. The high resolution image of the aqueous solution of the carbon quantum dots was measured, and the measured results are shown in fig. 2, and as can be seen from fig. 2, the lattice fringes of the carbon quantum dots are clearly visible. XPS elemental analysis data of the prepared aqueous carbon quantum dot solution were measured, and the results are shown in FIG. 3. In addition, the infrared absorption, ultraviolet-visible, excitation emission, different excitation light and fluorescence lifetime spectra of the prepared aqueous solution of the carbon quantum dots are measured, and the measured results are sequentially shown in fig. 4-8. From fig. 3 and fig. 4, it can be seen that the surface of the carbon quantum dot contains rich functional groups related to nitrogen and oxygen. From FIG. 5, it can be seen that pi-pi exists in the castor seed carbon quantum dots * (at 282 nm) and n-pi * Photon energy absorption (at 328 nm), i.e., the creation of an inverted bond orbital of the conjugated pi domain; it can be seen from fig. 6 that the fluorescence of the aqueous solution of carbon quantum dots is blue under the excitation of ultraviolet light; as can be seen from FIG. 7, the carbon quantum dots obtained in this example are excitedA dependency phenomenon; as can be seen from fig. 8, the fluorescence lifetime of the carbon quantum dots obtained in this example is in the nanometer scale.
In addition, 11 samples with pH values of 2-12 are respectively prepared by concentrated hydrochloric acid, sodium hydroxide solid and deionized water, 0.1g of the carbon quantum dot powder prepared in the embodiment is respectively added into the 11 samples, and the measured fluorescence intensity changes are shown in FIG. 9, and as can be seen from FIG. 9, the fluorescence intensity changes of the carbon quantum dots prepared in the embodiment are not large at different pH values.
Example 3 preparation of fluorescent carbon Quantum dots Using Castor seeds as carbon Source (method 2)
3g of castor seed powder prepared in the example 1 is taken and dispersed into 50ml of deionized water, stirred for 15min, and put into a hydrothermal kettle taking tetrafluoroethylene as an inner container for hydrothermal reaction, wherein the temperature of the hydrothermal reaction is 150 ℃ and the reaction time is 7h; filtering with a 0.45 mu m filter head after the reaction is finished, centrifuging at 7000rpm for 15min in a centrifuge, and concentrating the supernatant in a rotary steaming instrument at 70 ℃ to 35 mg/ml; adding the concentrated aqueous solution into column chromatography filled with 200-mesh silica gel powder, washing impurities with dichloromethane with purity of more than or equal to 99.0%, eluting with deionized water to obtain pure carbon quantum dot solution, and finally placing the pure carbon quantum dot solution into a freeze dryer, and drying at 40pa for 24h to obtain white castor seed carbon quantum dot powder. The aqueous solution of carbon quantum dots prepared in this example and the characterization at different pH were the same as in example 2.
Example 4 preparation of fluorescent carbon Quantum dots Using Castor seeds as carbon Source (method 3)
Dispersing 4g of the castor seed powder prepared in the example 1 into 50ml of deionized water, stirring for 20min, and putting into a hydrothermal kettle taking tetrafluoroethylene as an inner container for hydrothermal reaction, wherein the temperature of the hydrothermal reaction is 160 ℃, and the reaction time is 6h; filtering with a 0.22 mu m filter head after the reaction is finished, centrifuging at 8000rpm for 10min in a centrifuge, and concentrating the supernatant in a rotary steaming instrument at 75 ℃ to 30 mg/ml; adding the concentrated aqueous solution into column chromatography filled with 200-mesh silica gel powder, washing impurities with dichloromethane with purity of more than or equal to 99.0%, eluting with deionized water to obtain pure carbon quantum dot solution, and finally placing the pure carbon quantum dot solution into a freeze dryer, and drying at 30pa for 30h to obtain white castor seed carbon quantum dot powder. The aqueous solution of carbon quantum dots prepared in this example and the characterization at different pH were the same as in example 2.
Example 5 preparation of fluorescent carbon Quantum dots Using Castor seeds as carbon Source (method 4)
Dispersing 5g of the castor seed powder prepared in the example 1 into 50ml of deionized water, stirring for 20min, and putting into a hydrothermal kettle taking tetrafluoroethylene as an inner container for hydrothermal reaction, wherein the temperature of the hydrothermal reaction is 170 ℃, and the reaction time is 8h; filtering with a 0.22 mu m filter head after the reaction is finished, centrifuging at 8000rpm for 20min in a centrifuge, and concentrating the supernatant in a rotary steaming instrument at 80 ℃ to 40 mg/ml; adding the concentrated aqueous solution into a column chromatograph filled with 100-mesh silica gel powder, washing impurities with dichloromethane with purity of more than or equal to 99.0%, eluting with deionized water to obtain pure carbon quantum dot solution, and finally placing the pure carbon quantum dot solution into a freeze dryer, and drying at 30pa for 28h to obtain white castor seed carbon quantum dot powder. The aqueous solution of carbon quantum dots prepared in this example and the characterization at different pH were the same as in example 2.
Claims (8)
1. The preparation method of the fluorescent carbon quantum dot by taking castor seeds as a carbon source is characterized by comprising the following steps of:
a. dispersing peeled castor seed powder into deionized water, uniformly stirring, and placing into a hydrothermal kettle to perform hydrothermal reaction in a drying oven;
b. taking out the product after the reaction is finished, filtering and centrifuging, taking out the supernatant, and concentrating by rotary evaporation;
c. adding the concentrated aqueous solution into a column chromatography filled with silica gel powder, washing impurities with dichloromethane, extracting carbon quantum dots of castor seeds with deionized water, and freeze-drying to obtain powder.
2. The method for preparing fluorescent carbon quantum dots by taking castor seeds as a carbon source according to claim 1, wherein in the step a, the dosage ratio of castor seed powder to deionized water is 2-5 g:50ml.
3. The method for preparing fluorescent carbon quantum dots by using castor seeds as a carbon source according to claim 1, wherein the peeled castor seed powder in the step a is prepared by the following steps: and soaking the castor seeds in deionized water for 5-10 hours, peeling, grinding, sieving, and drying at 60-80 ℃ for 15-30 minutes.
4. The method for preparing the fluorescent carbon quantum dots by taking castor seeds as a carbon source according to claim 3, wherein the sieving aperture of the castor seeds after grinding is 100-200 meshes.
5. The method for preparing fluorescent carbon quantum dots by using castor seeds as a carbon source according to claim 1, wherein the hydrothermal kettle in the step a uses polytetrafluoroethylene as an inner container.
6. The method for preparing fluorescent carbon quantum dots by using castor seeds as a carbon source according to claim 5, wherein in the step a, the hydrothermal reaction temperature is 150-180 ℃ and the reaction time is 4-8 h.
7. The method for preparing the fluorescent carbon quantum dots by taking castor seeds as a carbon source according to claim 1, wherein in the step b, the aperture of a filter head is 0.22 mu m or 0.45 mu m, the rotation speed of a centrifugal machine is 6000-8000 rpm, the centrifugation time is 10-15 min, the rotary evaporation temperature of supernatant fluid is 65-80 ℃, and the rotary evaporation concentration is 30-40 mg/ml.
8. The method for preparing the fluorescent carbon quantum dots by taking castor seeds as a carbon source according to claim 1, wherein in the step c, the particle size of silica gel powder is 100-200 meshes, the purity of dichloromethane is more than or equal to 99.0%, the pressure in a freeze dryer cavity is 30-100pa, and the freeze drying time is 24-30 h.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110217721A1 (en) * | 2010-03-08 | 2011-09-08 | Afreen Allam | Water soluble fluorescent quantum carbon dots |
| CN106829917A (en) * | 2015-12-04 | 2017-06-13 | 中国科学院大连化学物理研究所 | A kind of carbon quantum dot and its application |
| CN106916590A (en) * | 2015-12-28 | 2017-07-04 | 江南大学 | A kind of method that fluorescent carbon point is prepared by waste material pumpkin seeds shell |
| CN109110747A (en) * | 2018-11-06 | 2019-01-01 | 湖南农业大学 | It is a kind of using extract oil residue as raw material prepare fluorescent carbon point method and its manufactured fluorescent carbon point |
| CN110697681A (en) * | 2019-10-12 | 2020-01-17 | 青海大学 | Method for preparing carbon dots from broad beans, carbon dots and application thereof |
| CN113025318A (en) * | 2021-02-02 | 2021-06-25 | 南京师范大学 | Carbon quantum dot using pepper as carbon source and preparation method and application thereof |
-
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- 2023-01-16 CN CN202310062865.XA patent/CN116103042A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110217721A1 (en) * | 2010-03-08 | 2011-09-08 | Afreen Allam | Water soluble fluorescent quantum carbon dots |
| CN106829917A (en) * | 2015-12-04 | 2017-06-13 | 中国科学院大连化学物理研究所 | A kind of carbon quantum dot and its application |
| CN106916590A (en) * | 2015-12-28 | 2017-07-04 | 江南大学 | A kind of method that fluorescent carbon point is prepared by waste material pumpkin seeds shell |
| CN109110747A (en) * | 2018-11-06 | 2019-01-01 | 湖南农业大学 | It is a kind of using extract oil residue as raw material prepare fluorescent carbon point method and its manufactured fluorescent carbon point |
| CN110697681A (en) * | 2019-10-12 | 2020-01-17 | 青海大学 | Method for preparing carbon dots from broad beans, carbon dots and application thereof |
| CN113025318A (en) * | 2021-02-02 | 2021-06-25 | 南京师范大学 | Carbon quantum dot using pepper as carbon source and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 武文波等: "碳量子点/曙红Y比率型荧光探针测定L-抗坏血酸", 《发光学报》, vol. 41, pages 331 - 338 * |
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