CN110964521B - Fluorescence quenching carbon dot for circularly detecting oxygen, test strip and application - Google Patents
Fluorescence quenching carbon dot for circularly detecting oxygen, test strip and application Download PDFInfo
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- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/65—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing carbon
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y20/00—Nanooptics, e.g. quantum optics or photonic crystals
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
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- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B32/00—Carbon; Compounds thereof
- C01B32/15—Nano-sized carbon materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
Abstract
The invention provides a fluorescence quenching carbon dot for circularly detecting oxygen, a test strip and application. When the carbon dots are exposed to air or oxygen, the fluorescence of the fluorescence quenching carbon dots can be restored to the original intensity, and the process can be repeatedly cycled. The method for detecting oxygen is simple to operate, easily available in raw materials, green, environment-friendly, rapid and sensitive, and is the first application of detecting oxygen in the field of carbon dots.
Description
Technical Field
The invention relates to the field of application of nano materials, in particular to a fluorescence quenching carbon dot for circularly detecting oxygen, a test strip and application.
Background
Oxygen is one of the most important chemicals on earth, and animal and plant respiration, combustion and various oxidation processes all need the participation of oxygen. The determination of molecular oxygen is of great significance to environmental chemistry, biomedicine and industry. In some cases, the presence of oxygen can cause negative effects, and visual detection of oxygen without the aid of sophisticated instrumentation is desirable.
Carbon Dots (CDs), a novel type of photoluminescent nanomaterial, have received great attention in recent years. The carbon dots have the advantages of simple raw materials, environmental friendliness, strong light stability, biocompatibility, low toxicity and the like, and have wide application and huge development potential in the aspects of fluorescence sensing, biological imaging, drug delivery, photoelectrochemistry and the like.
Compared with methods for detecting oxygen such as a gas chromatography method, an austenite analyzer method, an electrochemical sensing method and the like, the carbon dot nano material based on fluorescence recovery has the advantages of low price, simplicity in operation, obvious phenomenon and the like. At present, the detection of oxygen by using carbon points is not reported, and the carbon points have great application potential in the aspects of intracellular oxygen sensing, in-vivo thrombus treatment and the like based on the excellent properties of the carbon points.
Disclosure of Invention
The invention provides a fluorescence quenching carbon dot for circularly detecting oxygen, a test strip and application. The invention is also applied to test paper and a packaging bag, so that the detection of oxygen is more convenient, rapid and visual.
The technical scheme for realizing the invention is as follows:
a fluorescence quenching carbon spot for circularly detecting oxygen is prepared by the following steps:
(1) dissolving solid carbon point p-CDs in an organic solvent, and performing ultrasonic dissolution to obtain a solution A;
(2) dissolving the solution A obtained in the step (1) in a PBS buffer solution to obtain a solution B;
(3) placing the solution B obtained in the step (2) in a vacuum cuvette at room temperature, introducing nitrogen to remove original oxygen in the cuvette and the solution B, and creating an oxygen-free environment;
(4) injecting the solution B in the step (3) into the vacuum cuvette in the step (2) by using a micro-injector, wherein the solution B is in Na2S2O4Quenching red fluorescence under the action of the (3) to obtain a fluorescence quenching carbon dot solution, wherein the fluorescence quenching state can be maintained in an oxygen-free environment.
The solid Carbon Dots p-CDs were prepared according to the method described in Red, Green, and Blue Luminescence by Carbon Dots, Full-Color Emission Tuning and multicolor cellular Imaging, and the specific steps were as follows: 0.2g of p-phenylenediamine is dissolved in 20mL of ethanol and reacted for 12 hours at 180 ℃ by a hydrothermal method. Purifying by silica gel column chromatography, and rotary evaporating to obtain p-CDs solid.
The organic solvent in the step (1) is ethanol, glycol, methanol, acetonitrile, ethyl acetate, dimethyl sulfoxide or N, N-dimethylaniline; the concentration of the solution A is 1-2.5 mg/mL.
The pH value of the PBS buffer solution in the step (2) is 7.4, and the concentration of the solution B is 1.0-50.0 mu g/mL.
Na in the step (4)2S2O4The concentration of the solution is 10-50 mmol/L.
A fluorescence quenching carbon dot test strip for circularly detecting oxygen is prepared by the following steps:
(a) preparing solution A, solution B and Na according to the steps (1) and (2)2S2O4A solution;
(b) immersing the test paper into the solution B, and then completely drying to obtain test paper C;
(c) mixing Na2S2O4And (3) dropping the solution on the test paper C, observing under a 365nm ultraviolet lamp, and determining that the red fluorescence on the test paper C is completely quenched to obtain the fluorescence quenching carbon dot test paper.
Na in said step (c)2S2O4Upon dropping the solution on test paper C, the fluorescence began to quench and was completely quenched within 30 seconds.
And (4) at room temperature, opening the vacuum cuvette cover in the step (3) to allow oxygen to enter, placing the fluorescence quenching carbon point in an aerobic environment, and observing that the quenched red fluorescence is gradually recovered under the detection of a 365nm ultraviolet lamp or a fluorimeter.
Quenching of the fluorescence quenching carbon spot and detection of oxygen can be cycled repeatedly.
Placing the fluorescence quenching carbon dot test strip in an oxygen-free environment for a period of time at room temperature; then placing in air, contacting with oxygen, observing under 365nm ultraviolet lamp, and gradually recovering red fluorescence.
The fluorescence quenching carbon dot test strip is placed in an anaerobic environment for 0-24 hours and then placed in the air, and the fluorescence is recovered to the maximum value within 30 min.
The invention has the beneficial effects that: the invention uses simple and easily synthesized red light carbon dots and a cheap reducing agent Na2S2O4And reducing the carbon dots to completely quench the red fluorescence, thereby obtaining fluorescence quenching carbon dots. Under the oxygen-free environment, the state of fluorescence quenching can be maintained all the time. The red fluorescence recovered upon exposure to an aerobic environment. The method for detecting oxygen is simple to operate, and raw materials are easy to obtainThe method is rapid and sensitive, and the carbon dots can be recycled. Based on the excellent properties, the method can be applied to oxygen detection in vitro and has application potential in cells.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a graph showing the fluorescence quenching and fluorescence recovery of p-CDs in example 1.
FIG. 2 is a graph showing the fluorescence quenching and fluorescence recovery ultraviolet absorption spectra of p-CDs and p-CDs in example 1.
FIG. 3 shows the p-CDs in Na in example 12S2O4And fluorescence intensity at 615nm with seven repeated treatments of oxygen.
FIG. 4 is a photograph of the test paper of example 4 under a 365nm UV lamp in the absence of oxygen (left) and in the presence of oxygen (right).
FIG. 5 is a photograph of the test paper in the sealed packaging bag (left) and the opened packaging bag (right) under 365nm ultraviolet lamp in example 5.
Detailed Description
The technical solutions of the present invention will be described clearly and completely below with reference to embodiments of the present invention, and it should be apparent that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without inventive effort based on the embodiments of the present invention, are within the scope of the present invention.
Example 1
The method for circularly detecting oxygen by using the fluorescence quenching carbon dots comprises the following steps:
the prepared p-CDs are dissolved in ethanol to be completely dissolved, and the concentration is 1.0 mg/mL. The ethanol solution of p-CDs was dissolved in PBS buffer solution at pH 7.4 to a final concentration of 1.0. mu.g/mL.
2mL of the solution was placed in a vacuum cuvette and nitrogen was introduced to create an oxygen-free environment. Fresh preparation concentration of 10mmol/L Na2S2O4The solution was 4. mu.L of Na by using a micro syringe2S2O4The solution was poured into a vacuum cuvette and the fluorescence quenched. The nitrogen was stopped, the lid of the vacuum cuvette was opened and the fluorescence resumed. Introducing nitrogen gas, injecting Na2S2O4The solution, the fluorescence quenched, the nitrogen turned off, and the cap opened. This process was repeated seven times in total. When the fluorescence intensity is measured, the measurement is carried out by a fluorescence spectrophotometer. The excitation light was 496nm, and the fluorescence intensity in the range of 510nm to 700nm was collected.
As shown in fig. 1, the emission spectra detected on the fluorescence spectrophotometer were normalized. The excitation wavelength was 496nm, line 1 was the fluorescence intensity of the p-CDs solution itself, and line 2 was Na added2S2O4Post fluorescence quenching, line 3 is contact O2And the fluorescence is recovered.
As shown in fig. 2, the absorption spectra detected on the uv-vis spectrophotometer were normalized. Line 1 is the absorption curve of the p-CDs solution itself and line 2 is the addition of Na2S2O4Absorption curves for post fluorescence quenching detection, line 3 is contact O2Absorption curve detected after recovery of the post-fluorescence.
As shown in FIG. 3, Na was sequentially added to the p-CDs solution in example 12S2O4And O2The fluorescence intensity at 615nm was repeatedly measured 7 times.
Example 2
The method for circularly detecting oxygen by using the fluorescence quenching carbon dots comprises the following steps:
the prepared p-CDs are dissolved in ethanol to be completely dissolved, and the concentration is 2.0 mg/mL. The p-CDs ethanol solution was dissolved in PBS buffer solution at pH 7.4 to a final concentration of 25.0. mu.g/mL.
2mL of the solution was placed in a vacuum cuvette and nitrogen was introduced to create an oxygen-free environment. Fresh preparation concentration of 40mmol/L Na2S2O4The solution was then prepared by taking 5. mu.L of Na with a micro-syringe2S2O4The solution was poured into a vacuum cuvette and the fluorescence quenched. The nitrogen was stopped, the lid of the vacuum cuvette was opened and the fluorescence resumed. Introducing nitrogen gas, injecting Na2S2O4The solution, the fluorescence quenched, the nitrogen turned off, and the cap opened. This process was repeated three times in total. When the fluorescence intensity is measured, the measurement is carried out by a fluorescence spectrophotometer. The excitation light was 496nm, and the fluorescence intensity was collected in the range of 510nm to 700 nm.
Example 3
The method for circularly detecting oxygen by using the fluorescence quenching carbon dots comprises the following steps:
the prepared p-CDs are dissolved in ethanol to be completely dissolved, and the concentration is 2.5 mg/mL. The ethanol solution of p-CDs was dissolved in PBS buffer solution at pH 7.4 to a final concentration of 50.0. mu.g/mL.
2mL of the solution was placed in a vacuum cuvette and nitrogen was introduced to create an oxygen-free environment. Fresh preparation concentration of 50mmol/L Na2S2O4The solution was then treated with 10. mu.L of Na using a micro-syringe2S2O4The solution was poured into a vacuum cuvette and the fluorescence quenched. The nitrogen was stopped, the lid of the vacuum cuvette was opened and the fluorescence resumed. Introducing nitrogen gas, injecting Na2S2O4The solution, the fluorescence quenched, the nitrogen turned off, and the cap opened. This process was repeated three times in total. When the fluorescence intensity is measured, the fluorescence intensity is measured with a fluorescence spectrophotometer. The excitation light was 496nm, and the fluorescence intensity in the range of 510nm to 700nm was collected.
Example 4
The method for detecting oxygen by using fluorescence quenching carbon dot test paper comprises the following steps:
a series of cut circular filters were immersed in p-CDs aqueous solution at a concentration of 50.0. mu.g/mL and uniform red fluorescence was observed on the filters under a 365nm UV lamp. The filter paper is completely dried, and the p-CDs test paper is manufactured. Na with the concentration of 10.0 mu g/mL is dripped on the test paper2S2O4Solution, under 365nm ultraviolet lamp, determining complete quenching of red fluorescence, and preparing by fluorescence quenching carbon dot test paperThe process is completed. Then the test paper is put into a vacuum cuvette filled with nitrogen, and the test paper is placed for 15min in an oxygen-free environment. The nitrogen was then stopped and the cuvette lid opened to expose the test strip to oxygen. And when the test paper is observed under a 365nm ultraviolet lamp, the red fluorescence on the test paper is gradually recovered. After 15min, the fluorescence intensity returned to a maximum value.
As shown in FIG. 4, the fluorescence quenching carbon dot test paper was kept in a red fluorescence quenching state (left) under the irradiation of a 365nm ultraviolet lamp while nitrogen gas was passed through the paper. The cuvette lid was opened and oxygen was exposed and the red fluorescence on the test paper gradually recovered (right).
Example 5
The method for detecting oxygen in the packaging bag by using the fluorescence quenching carbon dot test paper comprises the following steps:
a series of cut square absorbent paper was immersed in p-CDs aqueous solution at a concentration of 50.0. mu.g/mL and uniform red fluorescence was observed on the absorbent paper under a 365nm UV lamp. And completely drying the absorbent paper, and finishing the preparation of the p-CDs test paper. Na with the concentration of 10.0 mu g/mL is dripped on the test paper2S2O4And (5) determining that the red fluorescence is completely quenched under a 365nm ultraviolet lamp, and manufacturing the fluorescence quenching carbon dot test paper. And then putting the test paper into a packaging bag, vacuumizing, sealing and placing for 2 hours, and then cutting off a vacuum bag to enable the test paper to be in contact with oxygen. And when the test paper is observed under a 365nm ultraviolet lamp, the red fluorescence on the test paper is gradually recovered. After 30min, the fluorescence intensity returned to a maximum value.
As shown in FIG. 5, the test paper in the well-sealed vacuum packaging bag is in a state of no red fluorescence under the irradiation of 365nm ultraviolet lamp (left). The test paper in the open package had a clear red fluorescence (right).
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A fluorescence quenching carbon spot for circularly detecting oxygen is characterized by comprising the following preparation steps:
(1) dissolving solid carbon dots p-CDs in an organic solvent, and performing ultrasonic dissolution to obtain a solution A;
(2) dissolving the solution A obtained in the step (1) in a PBS buffer solution to obtain a solution B;
(3) placing the solution B obtained in the step (2) in a vacuum cuvette at room temperature, introducing nitrogen to remove oxygen, and creating an oxygen-free environment;
(4) adding Na into the solution B in the step (3)2S2O4And (4) dissolving, and reacting to obtain the fluorescence quenching carbon dots.
2. The fluorescence quenching carbon spot for the cyclic detection of oxygen according to claim 1, characterized in that: the organic solvent in the step (1) is ethanol, glycol, methanol, acetonitrile, ethyl acetate, dimethyl sulfoxide or N, N-dimethylaniline; the concentration of the solution A is 1-2.5 mg/mL.
3. The fluorescence quenching carbon spot for the cyclic detection of oxygen according to claim 1, characterized in that: the pH value of the PBS buffer solution in the step (2) is 7.4, and the concentration of the solution B is 1.0-50.0 mu g/mL.
4. The fluorescence quenching carbon spot for the cyclic detection of oxygen according to claim 1, characterized in that: na in the step (4)2S2O4The concentration of the solution is 10-50 mmol/L.
5. A fluorescence quenching carbon dot test strip for circularly detecting oxygen is characterized by comprising the following preparation steps:
(a) preparing solution A, solution B and Na according to steps (1) and (2) of any one of claims 1 to 42S2O4A solution;
(b) immersing the test paper into the solution B, and then completely drying to obtain test paper C;
(c) mixing Na2S2O4Dropping the solution on the test paper C, observing under 365nm ultraviolet lamp, and determining red color on the test paper CAnd (4) completely quenching the fluorescence to obtain the fluorescence quenching carbon dot test strip.
6. The fluorescence quenching carbon dot test strip for circularly detecting oxygen according to claim 5, which is characterized in that: na in said step (c)2S2O4Upon dropping the solution on test paper C, the fluorescence began to quench and was completely quenched within 30 seconds.
7. Use of a fluorescence quenching carbon dot according to any one of claims 1 to 4 for the cyclic detection of oxygen, characterized in that: the fluorescence quenching carbon spot was placed in an aerobic environment and under detection of a 365nm ultraviolet lamp or a fluorimeter, a gradual recovery of the quenched red fluorescence was observed.
8. Use according to claim 7, characterized in that: quenching of the fluorescence quenching carbon spot and detection of oxygen can be cycled repeatedly.
9. The application of the fluorescence quenching carbon dot test strip in the circular detection of oxygen, which is characterized in that: placing the fluorescence quenching carbon dot test strip in an oxygen-free environment for a period of time at room temperature; then placing in air, contacting with oxygen, observing under 365nm ultraviolet lamp, and gradually recovering red fluorescence.
10. The application of the fluorescence quenching carbon dot test strip in the circulating detection of oxygen according to claim 9, characterized in that: the fluorescence quenching carbon dot test strip is placed in an anaerobic environment for 0-24 hours and then placed in the air, and the fluorescence is recovered to the maximum value within 30 min.
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