CN111978958B - Sea urchin-shaped carbon-based nano material and preparation method and application thereof - Google Patents
Sea urchin-shaped carbon-based nano material and preparation method and application thereof Download PDFInfo
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- CN111978958B CN111978958B CN202010830198.1A CN202010830198A CN111978958B CN 111978958 B CN111978958 B CN 111978958B CN 202010830198 A CN202010830198 A CN 202010830198A CN 111978958 B CN111978958 B CN 111978958B
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention discloses a sea urchin-shaped carbon-based nano material and a preparation method thereof. The preparation method comprises the following steps: adding uric acid and L-cysteine into an ethanol water solution according to the mass ratio of 1-2: 1, stirring to form a uniform suspension, reacting the suspension at 160-240 ℃ for 2-8 hours, cooling to room temperature, centrifuging to remove large particles, dialyzing, and drying to obtain the carbon-based nano material. The carbon-based nano material or the water solution thereof can be matched with a phosphate buffer solution to realize the quantitative detection of hypochlorite in different solutions; the water solution can also be uniformly dripped on chromatographic paper, and the paper-based detection chip is prepared after drying and is used for detecting hypochlorite. The sea urchin-shaped carbon-based fluorescent nano material is prepared by a one-step solvothermal method, the method is simple, easy to separate, high in yield, high in purity and good in selectivity, and the defects that the conventional hypochlorite probe molecule is long in synthesis and purification steps, low in yield and the like are overcome.
Description
Technical Field
The invention relates to the field of nano material preparation and analysis and detection, in particular to a sea urchin-shaped carbon-based nano material and a preparation method and application thereof.
Background
Hypochlorite, a representative active oxygen substance, is involved in various physiological and pathological processes and reacts with biomolecules such as cholesterol, protein, DNA, RNA, etc. Abnormal levels of hypochlorite are likely to cause cell damage and tissue destruction, ultimately leading to some irreversible disease. In addition, hypochlorite is also commonly used in the pretreatment process of drinking water due to anti-infection capability, and hypochlorite with too high concentration can generate oxidation reaction with specific organic matters, thus being unfavorable for human health. Therefore, it is very important to develop hypochlorite probes with high sensitivity and specificity.
Common instrumental detection methods include an electrochemical method, an electrochemiluminescence method, a precipitation titration method, a colorimetric method and the like, but the methods have certain defects, such as complicated sample preparation process, long detection time, insufficient selectivity and sensitivity and the like. The fluorescence analysis method has simple experimental steps, easy operation and high sensitivity, thereby being widely applied.
The fluorescent carbon-based nano material is a novel material, has the advantages of good light stability, easy surface modification, good biocompatibility and the like, and is well applied to the fields of biomolecule detection, environmental analysis and the like as a fluorescent probe. The luminescent carbon-based material with unique morphology is very helpful for improving the specificity of the detection method. The existing hypochlorite fluorescence detection method is mostly based on the fluorescence quenching phenomenon, so that the detection sensitivity is limited. Namely, the existing hypochlorite fluorescence detection has the defects of low sensitivity, poor specificity, complex fluorescent probe preparation process and the like.
Disclosure of Invention
The invention aims to provide a preparation method of a sea urchin-shaped carbon-based nano material.
The specific technical scheme is as follows:
a preparation method of a sea urchin-shaped carbon-based nano material comprises the following steps: adding uric acid and L-cysteine into an ethanol water solution according to the mass ratio of 1-2, stirring to form a uniform suspension, reacting the suspension at 160-240 ℃ for 2-8 h, cooling to room temperature, centrifuging to remove large particles, dialyzing, collecting the obtained solution, and drying to obtain the sea urchin-shaped carbon-based nano material.
Preferably, the ethanol aqueous solution is prepared by mixing ethanol and water according to a volume ratio of 1-2.
Preferably, the concentrations of the uric acid and the L-cysteine in the ethanol water solution are respectively 10.71-13.40 g/L and 5.35-10.72 g/L.
Preferably, the rotation speed of the centrifugation is 7000-15000r/min, and the time of the centrifugation is 10-30 min.
Preferably, the molecular weight cut-off for dialysis is 1000-3000Da, and the dialysis time is 10-15 h.
Preferably, the drying is rotary steaming drying at 50-60 ℃ for 1-3 h, or vacuum drying at 40-60 ℃ for more than 24 h.
The second purpose of the invention is to provide a sea urchin-shaped carbon-based nano material.
The specific technical scheme is as follows:
a carbon-based nanomaterial in the shape of sea urchin is prepared by the above preparation method.
The third purpose of the invention is to provide the application of the sea urchin-shaped carbon-based nano material.
The specific technical scheme is as follows:
the urchin-shaped carbon-based nano material is applied to the detection of hypochlorite.
Preferably, the method applied above comprises the following steps: firstly, adding the urchin-shaped carbon-based nano material or the aqueous solution thereof, a phosphate buffer solution with the pH of 5-9 and hypochlorite aqueous solutions with different concentration gradients into a container, and fixing the volume by using secondary distilled water; and (3) swirling the obtained mixed solution for 2-5 minutes, incubating for 10-15 minutes, placing the mixed solution in a micro fluorescence cuvette, setting excitation and emission gratings to be 2nm by taking 350nm as an excitation wavelength, collecting a fluorescence emission spectrum, establishing a standard curve of the fluorescence intensity at the maximum emission peak and the hypochlorite concentration, and realizing quantitative detection of hypochlorite in different solutions by adopting the standard curve.
Preferably, the method of the above application comprises the steps of: preparing the sea urchin-shaped carbon-based nano material into an aqueous solution, uniformly dripping the aqueous solution on chromatographic paper, and drying at 40-60 ℃ to prepare a paper-based detection chip for detecting hypochlorite.
The invention has the following beneficial results:
(1) The sea urchin-shaped carbon-based fluorescent nano material is prepared by a one-step solvothermal method, the preparation method is simple, easy to separate, high in yield and high in purity, and the defects of long synthesis and purification steps, low yield and the like of the conventional hypochlorite probe molecule are overcome.
(2) The sea urchin-shaped carbon-based nano material can generate oxidation reaction with hypochlorite to inhibit the intramolecular charge transfer process, so that the phenomena of purple shift of emission wavelength and fluorescence enhancement are caused, and the problem of low sensitivity of the conventional fluorescence quenching type hypochlorite probe is solved. The linear range of detection is 0.1-200 mu M, and the detection limit is 30nM.
(3) The prepared sea urchin-shaped carbon-based nano material has higher selectivity on hypochlorite and is not interfered by other common substances.
(4) Compared with other luminescent carbon-based materials, the prepared sea urchin-shaped carbon-based nano material has better light stability, and can be conveniently used for qualitative analysis of hypochlorite after being manufactured into a solid paper-based detection chip.
Drawings
FIG. 1 is a transmission electron microscope image of the sea urchin-shaped carbon-based nanomaterial prepared in example 1.
Fig. 2 shows fluorescence emission wavelengths of the echinoid-shaped carbon-based nanomaterial prepared in example 2 at different excitation wavelengths.
FIG. 3 is a fluorescence response spectrum of the sea urchin-shaped carbon-based nanomaterial prepared in example 3 for hypochlorite of different concentrations; the upper right hand picture is the change in fluorescence color under 365nm ultraviolet light.
FIG. 4 is a graph showing the fluorescence intensity ratio I in example 3 401nm /I 436nm Linear relationship with hypochlorite concentration.
FIG. 5 is a diagram showing selective recognition and detection of the carbon-based nanomaterial of sea urchin shape obtained in example 4.
Fig. 6 is a qualitative detection diagram of hypochlorite by the paper-based detection chip based on the echinoid carbon-based nanomaterial constructed in example 4, and fig. 6A and 6B are fluorescence emission diagrams in the paper-based detection chip before and after hypochlorite is added, respectively.
Detailed Description
Example 1
0.15g of uric acid and 0.15g of L-cysteine were added to a mixed solution of 7mL of ethanol and 7mL of water, and stirred with a glass rod for about 15 minutes to form a uniform suspension. The resulting suspension was transferred to a 25mL stainless steel reaction vessel with a Teflon liner and reacted at 185 ℃ for 4.5 hours. After the reaction, the reaction mixture was cooled to room temperature and centrifuged at 10 000r/min for 15 minutes to remove large particles. Dialyzing for 12 hours by using a dialysis bag with the molecular weight cutoff of 1000Da, collecting the obtained solution, and carrying out rotary evaporation for 2 hours at 50 ℃ to obtain the sea urchin-shaped carbon-based nano material.
Taking part of the carbon-based nanomaterial to prepare 1.5mg/mL aqueous solution (i.e. aqueous solution of carbon-based nanomaterial), and storing in a refrigerator at 4 deg.C.
The aqueous solution of the carbon-based nanomaterial of echinoid shape was tested by TEM, and fig. 1 is a transmission electron microscope picture of the prepared carbon-based nanomaterial. As can be seen from FIG. 1, the fluorescent carbon-based nanomaterial is sea urchin-shaped and has a size of about 585nm.
Example 2
0.15g of uric acid and 0.075g of L-cysteine were added to a mixed solution of 7mL of ethanol and 7mL of water, and stirred with a glass rod for about 15 minutes to form a uniform suspension. The resulting suspension was transferred to a 25mL stainless steel reaction vessel with a Teflon liner and reacted at 200 ℃ for 5 hours. After the reaction, the reaction mixture was cooled to room temperature and centrifuged at 10 000r/min for 15 minutes to remove large particles. Dialyzing for 12 hours by a dialysis bag with the molecular weight cutoff of 1000Da, collecting the obtained solution, and carrying out rotary evaporation for 80 minutes at 60 ℃ to obtain the sea urchin-shaped carbon-based nano material.
Taking part of the carbon-based nanomaterial to prepare 1.5mg/mL aqueous solution (i.e. aqueous solution of carbon-based nanomaterial), and storing in a refrigerator at 4 deg.C.
The aqueous solution of the sea urchin-shaped carbon-based nano material is used for performance test, fig. 2 shows the emission spectra of the carbon-based nano material in the aqueous solution when the excitation wavelength is 280nm, 300nm, 320nm, 330nm, 340nm, 350nm, 360nm, 370nm, 380nm, 400nm, 420nm and 440nm, and the emission wavelength position can change along with the change of the excitation wavelength, and the optimal excitation wavelength is 350 nm.
Example 3
0.6g of uric acid and 0.6g of L-cysteine were added to a mixed solution of 28mL of ethanol and 28mL of water, and stirred with a glass rod for about 15 minutes to form a uniform suspension. The resulting suspension was transferred to a 100mL stainless steel reaction vessel with a Teflon liner and reacted at 200 ℃ for 5 hours. After the reaction, the reaction mixture was cooled to room temperature and centrifuged at 10 000r/min for 15 minutes to remove large particles. Dialyzing for 12 hours by a dialysis bag with the molecular weight cutoff of 1000Da, collecting the obtained solution, and carrying out rotary evaporation for 80 minutes at 60 ℃ to obtain the sea urchin-shaped carbon-based nano material.
Taking part of the carbon-based nanomaterial to prepare 1.5mg/mL aqueous solution (namely the aqueous solution of the carbon-based nanomaterial), and storing in a refrigerator at 4 ℃.
Putting 10 mu L of sea urchin-shaped carbon-based nano material aqueous solution into a 2mL centrifuge tube, adding 50 mu L of phosphate buffer (pH is 7.0), then adding hypochlorite aqueous solution with different concentration gradients, and fixing the volume to 500 mu L by using secondary distilled water; swirling the mixed solution for 2-5 minutes, and incubating for 10 minutes; placing in 700 μ L micro fluorescence cuvette, setting excitation and emission grating at 2nm with 350nm as excitation wavelength, and collecting fluorescence emission spectrum; and establishing a standard curve of the fluorescence intensity at the maximum emission peak and the hypochlorite concentration through the measured fluorescence emission spectrum to realize quantitative detection of the hypochlorite.
FIG. 3 shows that when the prepared urchin-shaped carbon-based nanomaterial is used to detect sodium hypochlorite with different concentrations, the fluorescence emission changes with the concentration of hypochlorite, and as can be seen from the upper right-hand graph in FIG. 3, after hypochlorite is added, under a 365nm ultraviolet lamp, the original blue color of the solution changes into purple, the fluorescence intensity gradually increases, and the fluorescence emission peak position is blue-shifted from 436nm to 401nm.
FIG. 4 is a graph showing the fluorescence intensity ratio I in example 3 401nm /I 436nm The linear relationship with hypochlorite concentration can be obtained by the method, the linear range of the detection method is 0.1-200 mu M, and the detection limit is 30nM.
Example 4
0.75g of uric acid and 0.45g of L-cysteine were added to a mixed solution of 28mL of ethanol and 28mL of water, and stirred with a glass rod for about 15 minutes to form a uniform suspension. The resulting suspension was transferred to a 100mL stainless steel reaction vessel with a Teflon liner and reacted at 200 ℃ for 8 hours. After the reaction, the reaction mixture was cooled to room temperature and centrifuged at 10 000r/min for 15 minutes to remove large particles. Dialyzing for 12 hours by a dialysis bag with the molecular weight cutoff of 1000Da, collecting the obtained solution, and drying in vacuum for 48 hours at 45 ℃ to obtain the sea urchin-shaped carbon-based nano material.
Taking part of the carbon-based nanomaterial to prepare 1.5mg/mL aqueous solution (i.e. aqueous solution of carbon-based nanomaterial), and storing in a refrigerator at 4 deg.C.
Putting 50 mu L of sea urchin-shaped carbon-based nano material aqueous solution into a 2mL centrifuge tube, adding 50 mu L of phosphate buffer solutions with different pH values of 7.0, and then adding different substance aqueous solutions (CuSO) 4 、ZnSO 4 、PbSO 4 、NiCl 2 、MnSO 4 、CoCl 2 、Hg(NO 3 ) 2 +EDTA、KI、NaBr、NaCl、NaF、NaNO 3 、NaNO 2 、Na 2 SO 4 、Na 3 PO 4 、Na 2 HPO 4 、Na 2 CO 3 、H 2 O 2 NaClO), and the volume is adjusted to 500 mu L by secondary distilled water; transferring to a 700-mu-L fluorescence cuvette, setting the excitation wavelength to be 350nm and the excitation and emission grating to be 2nm, and collecting the fluorescence emission spectrum. Fig. 5 is a selective recognition and detection diagram of the prepared sea urchin-shaped carbon-based nanomaterial, and it can be seen from the diagram that the sea urchin-shaped carbon-based nanomaterial only has a fluorescent response to hypochlorite.
Uniformly dripping 200 mu L of sea urchin-shaped carbon-based nano material aqueous solution on chromatographic paper, placing the chromatographic paper on an oven, and drying at 40-60 ℃ to obtain the paper-based detection chip. FIG. 6 is a qualitative detection diagram of hypochlorite by the paper-based detection chip, and it can be seen from FIG. 6 that the paper-based detection chip changes blue fluorescence (FIG. 6A) into purple fluorescence (FIG. 6B) after hypochlorite is added, thus confirming that the solid-phase urchin-shaped carbon-based nano material adsorbed on the paper fiber still has qualitative detection capability.
Claims (10)
1. A preparation method of a sea urchin-shaped carbon-based nano material is characterized by comprising the following steps: adding uric acid and L-cysteine into an ethanol water solution according to the mass ratio of 1-2, stirring to form a uniform suspension, reacting the suspension at 160-240 ℃ for 2-8 h, cooling to room temperature, centrifuging to remove large particles, dialyzing, collecting the obtained solution, and drying to obtain the sea urchin-shaped carbon-based nano material.
2. The method for preparing the sea urchin-shaped carbon-based nanomaterial according to claim 1, wherein the ethanol aqueous solution is prepared by mixing ethanol and water according to a volume ratio of 1-2.
3. The method for preparing the sea urchin-shaped carbon-based nanomaterial according to claim 1, wherein the concentrations of the uric acid and the L-cysteine in the ethanol aqueous solution are 10.71-13.40 g/L and 5.35-10.72 g/L, respectively.
4. The method for preparing the sea-urchin-shaped carbon-based nanomaterial according to claim 1, wherein the rotation speed of the centrifugation is 7000-15000r/min, and the time of the centrifugation is 10-30 min.
5. The method for preparing the sea urchin-shaped carbon-based nanomaterial according to claim 1, wherein the cut-off molecular weight of a dialysis bag used for dialysis is 1000-3000Da, and the dialysis time is 10-15 h.
6. The method for preparing the sea urchin-shaped carbon-based nanomaterial according to claim 1, wherein the drying is rotary evaporation drying at 50-60 ℃ for 1-3 h, or vacuum drying at 40-60 ℃ for more than 24 h.
7. A carbon-based nanomaterial in the shape of sea urchin, characterized by being prepared by the preparation method of any one of claims 1 to 6.
8. Use of a carbon-based nanomaterial of the shape of the sea urchin according to claim 7, in the detection of hypochlorite.
9. Use according to claim 8, characterized in that it comprises the following steps: firstly, adding the urchin-shaped carbon-based nano material or the aqueous solution thereof, a phosphate buffer solution with the pH of 5-9 and hypochlorite aqueous solutions with different concentration gradients into a container, and fixing the volume by using secondary distilled water; and (3) swirling the obtained mixed solution for 2-5 minutes, incubating for 10-15 minutes, placing in a micro-fluorescence cuvette, setting excitation and emission gratings to be 2nm by taking 350nm as an excitation wavelength, collecting a fluorescence emission spectrum, establishing a standard curve of fluorescence intensity at a maximum emission peak and hypochlorite concentration, and realizing quantitative detection of hypochlorite in different solutions by adopting the standard curve.
10. Use according to claim 8, characterized in that it comprises the following steps: preparing the sea urchin-shaped carbon-based nano material into an aqueous solution, uniformly dripping the aqueous solution on chromatographic paper, and drying at 40-60 ℃ to prepare a paper-based detection chip for detecting hypochlorite.
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| CN105319192A (en) * | 2015-11-12 | 2016-02-10 | 湖南科技大学 | Method for detecting hypochlorite anions through water-soluble fluorescent silica nanoparticle |
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| CN111286324A (en) * | 2020-03-27 | 2020-06-16 | 上海应用技术大学 | Fluorescent probe for detecting hypochlorite in water environment and preparation method and application thereof |
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| CN105319192A (en) * | 2015-11-12 | 2016-02-10 | 湖南科技大学 | Method for detecting hypochlorite anions through water-soluble fluorescent silica nanoparticle |
| CN110982516A (en) * | 2019-12-04 | 2020-04-10 | 盐城工学院 | Preparation method and application of fluorescent carbon-based nanobelt with narrow half-peak width |
| CN111286324A (en) * | 2020-03-27 | 2020-06-16 | 上海应用技术大学 | Fluorescent probe for detecting hypochlorite in water environment and preparation method and application thereof |
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| Strong enhancement of the chemiluminescence of hydrogen sulfite-oxidant systems in the presence of N,S-doped graphene quantum dots, and its application to the determination of folic acid in spinach and kiwifruit samples;Junmei Zhang等;《Food Analytical Methods》;20190104;第12卷;第869-876页 * |
| Sulfur and nitrogen co-doped carbon quantum dots as the chemiluminescence probe for detection of Cu2+ ions;Mohammad Amjadi等;《Journal of Luminescence》;20161020;第182卷;第246-251页 * |
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