CN107340278B - Method for detecting oxygen by adopting fluorescent molecular probe based on fullerene or fullerene derivative - Google Patents
Method for detecting oxygen by adopting fluorescent molecular probe based on fullerene or fullerene derivative Download PDFInfo
- Publication number
- CN107340278B CN107340278B CN201710419871.0A CN201710419871A CN107340278B CN 107340278 B CN107340278 B CN 107340278B CN 201710419871 A CN201710419871 A CN 201710419871A CN 107340278 B CN107340278 B CN 107340278B
- Authority
- CN
- China
- Prior art keywords
- fullerene
- oxygen
- solution
- fluorescent
- illumination
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
-
- 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
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
Landscapes
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pathology (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Optics & Photonics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a method for detecting oxygen by adopting a fluorescent molecular probe based on fullerene or a derivative thereof, belonging to the application field of fullerene materials. The method comprises the following steps: mixing fullerene or fullerene derivatives and fluorescent molecules to form fullerene solution containing the fluorescent molecules, vacuumizing, introducing an oxygen sample, generating singlet oxygen under the illumination condition, generating epoxy compounds by the fluorescent molecules, carrying out fluorescence quenching, detecting the intensity of fluorescent signals in a system, and further detecting the oxygen content in the system. The invention solves the problems that the existing oxygen detection has low precision and is difficult to detect trace oxygen. The fullerene or fullerene derivative can generate singlet oxygen under the illumination condition, the oxygen content can be indirectly detected according to the reduced fluorescence intensity, the detection sensitivity is high, the endoperoxide generated after the reaction can generate over 90 percent of reverse reaction, and the high recycling rate is achieved.
Description
Technical Field
The invention relates to the application field of fullerene materials, in particular to a method for detecting oxygen by adopting a fluorescent molecular probe based on fullerene or derivatives thereof.
Background
Oxygen plays a very important role in biological life activities. Cells within an organism require oxygen for aerobic metabolism to maintain daily physiological activities. Hypoxia can seriously impair the life activities of the living beings, and in serious cases, coma and even death can occur. Excess oxygen can cause "oxygen poisoning" which can lead to vertigo, dysesthesia, tingling, impaired vision and hearing, and hypoconsciousness. Only proper oxygen concentration can maintain the normal running of biological life activities. Therefore, it is very important to detect the oxygen content in the organism.
Currently, some oxygen detection methods such as titration, amperometry, and thermoluminescence are available. Although these methods can effectively detect the oxygen content, there are some disadvantages, such as: the response time is long, the continuous detection cannot be realized, the interference by other gases is easy to occur, the required cost is high, and the accurate observation and measurement of trace oxygen by an instrument are difficult.
Currently, fluorescent probes can be used to detect active oxygen. Singlet oxygen is an excited state of oxygen and is also a highly reactive molecule, also a typical reactive oxygen molecule. Most singlet oxygen fluorescent probes are based on its reaction with anthracene. Most singlet oxygen fluorescent probes select anthracene or 9, 10-diphenylanthracene as the recognition group. Under the action of singlet oxygen, the fluorescent probe generates cycloaddition reaction to generate endoperoxide, and the fluorescence of the fluorophore is quenched, thereby achieving the purpose of detecting singlet oxygen.
Therefore, oxygen can be converted into singlet oxygen under certain conditions, and the fluorescent molecular probe for detecting the oxygen concentration, which has the advantages of fast response time, good selectivity, high sensitivity and simple operation, is designed by means of the characteristic that fluorescence is obviously changed after the interaction of fluorescent molecules and singlet oxygen, and is expected to be widely applied to the fields of bioscience, basic medicine, environmental detection and the like.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a method for detecting oxygen by using a fluorescent molecular probe, and particularly provides a method for detecting oxygen by using a fluorescent molecular probe based on fullerene or fullerene derivatives.
The method for detecting oxygen by adopting the fluorescent molecular probe comprises the following steps:
(1) dissolving fullerene and/or fullerene derivatives in halogenated benzene, alkylbenzene or carbon disulfide to prepare a solution containing the fullerene and/or the fullerene derivatives, namely a solution 1;
(2) dissolving the fluorescent molecules in halogenated benzene, alkylbenzene or carbon disulfide to prepare a solution containing the fluorescent molecules, namely a solution 2;
(3) mixing the solution 1 and the solution 2 to obtain a fluorescent molecule solution containing fullerene and/or fullerene derivatives;
(4) vacuumizing the mixed solution obtained in the step (3) to reduce the oxygen concentration to 0ml/cm3Obtaining a detection system;
(5) under the illumination condition, establishing a linear relation between the fluorescence signal intensity and the oxygen content in a detection system;
(6) and (5) introducing the sample to be detected into the detection system same as the step (4), measuring the fluorescence signal intensity in the detection system under the same illumination condition as the step (5), and further calculating to obtain the oxygen content in the sample to be detected.
The fullerene and/or fullerene derivative can interact with oxygen with different concentrations to generate singlet oxygen under the illumination condition, the fluorescent molecule reacts with the singlet oxygen to generate an epoxy compound of the fluorescent molecule, namely an endoperoxide compound, and the epoxy compound has low fluorescence intensity or does not emit fluorescence, namely generates fluorescence quenching.
The reaction of the fluorescent molecule and singlet oxygen is a reversible reaction: due to the fact that the epoxy compound generated by the reaction of the fluorescent molecules and singlet oxygen is unstable, under the heating condition, the epoxy compound can perform reversible reaction and be changed into fluorescent molecules with high fluorescence intensity again under the conditions that the temperature range is 100-120 ℃ and the heating time is 20-60 min, and therefore the fluorescent probe can be recycled, and is efficient and convenient. Using the characteristic uv absorption of the fluorescent molecular bulk and the change in the characteristic uv absorption after the epoxy compound is generated, it can be found that almost 90% or more of the epoxy compound can undergo reversible reactions (see fig. 1).
The fullerene includes at least one of an empty fullerene and an endohedral fullerene.
The hollow fullerene comprises a general formula of C2mWherein m is not less than 30 and not more than 60, and optionally C60、C70、C76、C78、C82、C84At least one of (1).
The endohedral fullerene comprises N @ C60、La@C72、Sc2@C74、Sc2@C76、La2@C80、Sc3N@C80、Tm@C82、Gd@C82、Sc2C2@C84At least one of;
the fullerene derivative comprises at least one of fullerene oxide and fullerene addition product, and the optional fullerene oxide is C60O、C70O, and the fullerene addition product is at least one of fullerene 1, 3-dipolar cycloaddition products and fullerene binger addition products.
The fluorescent molecule is an organic molecule with a conjugated structure and a biphenyl structure, and can be selected from 9, 10-diphenylanthracene and 9-anthracene-beta-propionic acid.
The halogenated benzene is chlorobenzene, and the alkylbenzene is toluene.
The concentration of fullerene and/or fullerene derivative in the solution 1 is 10-6mol/L～10-2mol/L。
The concentration of the fluorescent molecules in the solution 2 is 10-6mol/L～10-2mol/L。
When the solution 1 and the solution 2 are mixed in the step (3), the volume ratio of the solution 1 to the solution 2 is 1: 3-3: 1.
The sample to be detected which is introduced can comprise a gaseous sample or a liquid sample.
The illumination conditions comprise xenon lamp illumination, halogen lamp illumination and mercury lamp illumination, and optionally xenon lamp illumination and further optionally xenon lamp white light source illumination.
The illumination intensity is 1 mw-30 mw, and the illumination time is 1 min-60 min.
The specific steps for establishing the linear relation between the fluorescence signal intensity and the oxygen content in the detection system in the step (5) are as follows: in the detection system, a standard curve graph with the abscissa as the oxygen concentration and the ordinate as the fluorescence signal intensity is drawn according to a plurality of oxygen concentrations and the fluorescence intensity corresponding to each oxygen concentration, and a linear range is determined.
Compared with the prior art, the invention has the following beneficial effects:
(1) the fullerene and/or fullerene derivative can interact with oxygen with different concentrations under the illumination condition,the detection of the change of ESR (electron spin resonance) electron spin resonance signals proves that the interaction can efficiently generate different amounts of singlet oxygen (1O2) And the final product obtained by the interaction of the fluorescent molecule and the singlet oxygen can be proved to be an epoxy compound through nuclear magnetic resonance hydrogen spectrum detection. Under the condition of illumination and under the condition that the fullerene and/or fullerene derivative and the fluorescent molecule are in sufficient quantity, the rates of the formation of singlet oxygen by oxygen and the reaction of the singlet oxygen and the fluorescent molecule are both very rapid, so that the oxygen concentration when the oxygen is detected firstly can be roughly considered as the lowest oxygen concentration which can be detected by the detection method. Under the experimental conditions of example 1, the decrease in fluorescence was detected after 2 seconds of oxygen gas introduction, at which time the oxygen concentration in the detection system was 0.0707ml/cm3The detection limit is low, and the detection sensitivity is high; under the experimental conditions of example 3, the fluorescence intensity of the detection system is obviously reduced after 2min of illumination, the reduction rate is about 89%, and the response speed of the detection method is high, so that the detection method is efficient and has strong operability.
(2) The types of the fullerene can be selected, and the conditions for preparation, separation and purification are mature.
(3) The selected fluorescent molecules have stable properties, low cost and obvious fluorescent characteristics.
(4) After the oxygen detection is finished, 90% of the obtained epoxy compound, namely endoperoxide, can perform reversible reaction under the heating condition and is changed into fluorescent molecules with large fluorescence intensity again, so that the fluorescent probe can be repeatedly utilized.
Drawings
FIG. 1 is a UV-VIS absorption spectrum of a reversible change in a 9, 10-diphenylanthracene fluorescent molecule.
FIG. 2 is a diagram showing the reaction process between the fluorescent probe 9, 10-diphenylanthracene and singlet oxygen in the presence of a hollow fullerene in example 1.
FIG. 3 shows the fluorescence probe 9, 10-diphenylanthracene1O2Nuclear magnetic resonance hydrogen spectra before and after the reaction.
FIG. 4 is a graph showing the change in ESR (electron spin resonance) electron spin resonance before and after the addition of TEMP (2,2,6, 6-tetramethylpiperidinol) as a capture agent to the detection system of example 1.
FIG. 5 is a fluorescence quenching spectrum of the detection system obtained in the detection result verification step of example 1 with different time illumination.
FIG. 6 is a diagram showing the reaction process between the fluorescent probe 9-anthracene- β -propionic acid of example 2 and singlet oxygen in the presence of a hollow fullerene.
FIG. 7 shows the fluorescence probe 9-anthracene-beta-propionic acid1O2Nuclear magnetic resonance hydrogen spectra before and after the reaction.
FIG. 8 is a fluorescence quenching spectrum of the detection system obtained in the detection result verification step of example 2 with different time illumination.
FIG. 9 is a diagram showing the reaction process between the fluorescent probe 9, 10-diphenylanthracene and singlet oxygen in the presence of endofullerene in example 3.
FIG. 10 is a fluorescence quenching spectrum of the detection system obtained in the detection result verification step of example 3 with different time illuminations.
FIG. 11 is a graph showing the decrease in fluorescence signal intensity under the conditions of example 1 for different oxygen contents.
Detailed Description
The following detailed description of the present invention is provided in conjunction with the accompanying drawings, but it should be understood that the scope of the present invention is not limited to the specific embodiments.
The following examples are given without specific reference, and the detection system is set up at 4cm3Wherein the detection system in the container is 3ml, that is, the fluorescent molecule solution containing fullerene and/or fullerene derivative is 3 ml.
The fluorescence intensity values in the detection systems of examples 1 to 3 were all read under the detection condition that the maximum absorption value was 445 nm.
Example 1
Based on fullerene C60The method for detecting oxygen by adopting the fluorescent molecular probe 9, 10-diphenylanthracene has a reaction scheme shown in figure 2, and comprises the following steps:
(1) reacting fullerene C60Is dissolved inPreparing fullerene C in carbon disulfide60At a concentration of 10-31, mol/L solution;
(2) dissolving fluorescent molecule 9, 10-diphenylanthracene in carbon disulfide to prepare 9, 10-diphenylanthracene with concentration of 10- 2mol/L solution 2;
(3) adding the solution 2 into the solution 1 according to the volume ratio of the solution 2 to the solution 1 of 1:1 to obtain the fullerene C60The fluorescent molecule solution of (1);
(4) vacuumizing the mixed solution obtained in the step (3) to reduce the oxygen concentration to 0ml/cm3Obtaining detection systems, and then respectively introducing pure oxygen with the volumes of 0ml, 0.555ml, 1.11ml, 1.665ml, 2.22ml, 2.775ml and 3.33ml into 7 independent detection systems, wherein the oxygen concentration in each detection system is 0ml/cm in sequence3、0.1388ml/cm3、0.2775ml/cm3、0.4163ml/cm3、0.555ml/cm3、0.6938ml/cm3、0.8325ml/cm3；
(5) Irradiating the detection system for more than 2min by using a 20mw xenon lamp white light source with the wavelength of 200-1000 nm (wherein the sample has the maximum absorption at 445 nm), generating singlet oxygen by oxygen in a closed container, generating an epoxy compound by fluorescent molecules and the singlet oxygen, carrying out fluorescence quenching, detecting the fluorescence signal intensity in the system, and drawing a standard curve graph with the abscissa as the oxygen concentration and the ordinate as the signal intensity as shown in figure 11.
Wherein, the specific process of oxygen introduction is as follows: oxygen was introduced into the closed vessel containing the above 3ml detection system through a small hole having a diameter of 0.3mm at a flow rate of 2 m/s.
When oxygen is introduced for 2s, the fluorescence can be detected to be reduced, which indicates that after the oxygen is introduced for 2s into the detection system, the epoxy compound begins to generate in the presence of fullerene, and the volume of the oxygen introduced correspondingly at this time is: (2 m/s.times.2sX.102)×(π×(0.3mm/2)2×10-2) 0.2826ml, and the oxygen concentration of the detection system at 2s inlet was obtained as: 0.2826/4-0.0707 ml/cm3。
As can be seen from FIG. 11, at an oxygen concentration of 0.707ml/cm3To 0.555ml/cm3In the range of (1), the fluorescence signal intensity shows a linear change with increasing oxygen concentration, and the obtained function is:
y＝3398.2-4173.8x
wherein x is the oxygen concentration and y is the fluorescence intensity
According to the formula, under the same condition (namely, the fullerene and the fluorescent molecule have the same type and content, and the light ray type and the irradiation time of the system to be detected are the same), the concentration of oxygen in the sample to be detected can be obtained according to the intensity value of the detected fluorescent signal of the sample to be detected, so that the oxygen content in the sample to be detected can be detected.
And (3) verifying the detection result: preparing a new closed container with the volume of 4ml, vacuumizing the detection system which is the same as that in the step (4), introducing 0.74ml of pure oxygen, and after illumination is carried out for 2min, detecting that the fluorescence intensity of the system is 2700 a.u.; the oxygen concentration was changed from 0.74/4 to 0.185ml/cm3And (3) substituting the linear function y in the step (5) into 3398.2-4173.8x to obtain the theoretical fluorescence intensity of 2626a.u., which is different from the detected fluorescence intensity of 2700a.u. by only 74a.u., within an error tolerance range, so that the detection method adopted by the invention can be used for detecting the specific content of oxygen in the sample to be detected. Therefore, the detection method can accurately detect the oxygen content of lower concentration.
In the nuclear magnetic resonance hydrogen spectrum of FIG. 3 (DMSO was used as a deuteration reagent), a, b, c, d, and e represent five hydrogens corresponding to 9, 10-diphenylanthracene, respectively, and a ', b ', c ', d ', and e ' represent five hydrogens of 9, 10-diphenylanthracene endoperoxide. The uppermost graph in fig. 3 represents the un-illuminated nuclear magnetic hydrogen spectrum of the uniform mixture of 9, 10-diphenylanthracene and fullerene, and it can be seen from the hydrogen spectrum that 9, 10-diphenylanthracene is not changed, which indicates that 9, 10-diphenylanthracene does not chemically react with fullerene under the un-illuminated condition; the following two figures respectively represent nuclear magnetic hydrogen spectrums obtained after oxygen is introduced into a solution in which 9, 10-diphenylanthracene and fullerene are uniformly mixed for 20s and 100s and the solution is irradiated for 2min under a xenon lamp light source, and it can be seen that new substances appear in the hydrogen spectrums, which indicates that the mixed solution is subjected to chemical reaction under the illumination condition. When the oxygen introduction amount is 20s, 9, 10-diphenylanthracene remains, and a hydrogen spectrum shows a mixture of 9, 10-diphenylanthracene and 9, 10-diphenylanthracene endoperoxide; when 100s is introduced, the 9, 10-diphenylanthracene is completely reacted, and all the products are 9, 10-diphenylanthracene endoperoxides. Comparing the two substances before and after the system is illuminated, the number of hydrogen is unchanged, but the relative displacement change occurs, which indicates that a new substance is generated, and comparing the position of hydrogen, the newly generated substance can be determined to be 9, 10-diphenylanthracene endoperoxide.
And detecting singlet oxygen by ESR electron spin resonance, wherein the detection process is as follows: due to TEMP capture1O2Has very strong capability of1O2Can be captured, so TEMP (2,2,6, 6-tetramethyl piperidinol) is selected as the capture agent, and the TEMP captures the TEMP in the detection system after being illuminated1O2The generation of TEMPOL (1-oxyl-2, 2,6, 6-tetramethyl-4-hydroxypiperidine) is detected, and the ESR signal of TEMPOL is obviously enhanced (see figure 4), which proves that the detection system generates singlet oxygen1O2Thereby proving in fullerene C60Under the action of the light, oxygen in the detection system is converted into singlet oxygen through illumination, so that the quenching of fluorescence is caused.
Meanwhile, as is clear from FIG. 5, the fluorescence intensity of the system before light irradiation was 3500a.u., and the fluorescence intensity of the system after light irradiation for 2min was 2700a.u., so that 9, 10-diphenylanthracene and fullerene C were contained60The fluorescence intensity of the detection system is obviously reduced after illumination, the reduction rate of the fluorescence intensity is 23 percent, and the response time is fast; meanwhile, the fluorescence signal intensity of the molecules is not obviously reduced along with the extension of the illumination time.
Description of the drawings: the fluorescence signal intensity in FIG. 5 is in the order of 9, 10-diphenylanthracene + C from top to bottom60(not illuminated), 9, 10-diphenylanthracene + C60(2 min of illumination), 9, 10-diphenylanthracene + C60(illumination for 4min), 9, 10-diphenylanthracene + C60(illumination for 6min), 9, 10-diphenylanthracene + C60(illumination for 8min), 9, 10-diphenylanthracene + C60(illumination for 10min), 9, 10-diphenylanthracene + C60(12 min of light irradiation) under the same conditions.
Example 2
Based on richLeen C60The method for detecting oxygen by adopting the fluorescent molecular probe 9-anthracene-beta-propionic acid has a reaction scheme as shown in figure 6, and comprises the following steps:
(1) reacting fullerene C60Dissolving in carbon disulfide to obtain fullerene C60At a concentration of 10-31, mol/L solution;
(2) dissolving fluorescent molecule 9-anthracene-beta-propionic acid in carbon disulfide to prepare 9-anthracene-beta-propionic acid with the concentration of 10- 4mol/L solution 2;
(3) according to the weight ratio of the solution 2: the volume ratio of the solution 1 is 1:1, the solution 1 is added into the solution 2 to obtain the fullerene C60The fluorescent molecule solution of (1);
(4) and (3) constructing 7 detection systems according to the step (4) in the embodiment 1, and simultaneously drawing a standard curve graph with the abscissa as the oxygen concentration and the ordinate as the signal intensity according to the step (5) in the embodiment 1 to further obtain a linear function of the fluorescence intensity and the oxygen concentration, so that the concentration of the oxygen in the sample to be detected can be obtained according to the intensity value of the detected fluorescence signal of the sample to be detected.
And (3) verifying the detection result: preparing a new closed container with the volume of 4ml, vacuumizing the detection system which is the same as that in the step (4), introducing 0.74ml of pure oxygen, and after illumination is carried out for 2min, detecting that the fluorescence intensity of the system is 2400 a.u.; meanwhile, the function obtained according to the step (4) and the oxygen concentration of 0.74/4-0.185 ml/cm3The theoretical fluorescence intensity was found to be 2320a.u. which differs from the detected fluorescence intensity 2400a.u. by only 80a.u., within the tolerance of error. Therefore, the detection method can accurately detect the oxygen content of lower concentration.
Nuclear magnetic resonance hydrogen spectrum of FIG. 7 (using CDCl)3As deuteration reagent), a, b, c, d, e and f respectively represent six hydrogen corresponding to 9-anthracene-beta-propionic acid, and a ', b', c ', d', e 'and f' represent six hydrogen of 9-anthracene-beta-propionic acid endoperoxide. The uppermost graph in FIG. 7 represents the un-illuminated nuclear magnetic hydrogen spectrum of the homogeneous mixture of 9-anthracene-beta-propionic acid and fullerene, and it can be seen from the hydrogen spectrum that 9-anthracene-beta-propionic acid is not changed, which shows that 9-anthracene-beta-propionic acid is not illuminatedThe acid does not react with the fullerene chemically; the following figure represents the nuclear magnetic hydrogen spectrum of the uniformly mixed solution of 9-anthracene-beta-propionic acid and fullerene after oxygen is introduced for 20s and the solution is illuminated for 2min under a xenon lamp light source, and it can be seen that new substances appear in the hydrogen spectrum, which indicates that a chemical reaction occurs in the mixed solution under the illumination condition. When oxygen was passed for 20s, 9-anthracene- β -propionic acid remained and the hydrogen spectrum shows a mixture of 9-anthracene- β -propionic acid and 9-anthracene- β -propionic acid endoperoxide. The hydrogen spectrum can be used for comparing two substances before and after the system is illuminated, the number of hydrogen does not change, but the relative displacement change occurs, the generation of a new substance is proved, and the newly generated substance can be determined to be 9-anthracene-beta-propionic acid endoperoxide compared with the position of hydrogen.
As can be seen from FIG. 8, the fluorescence intensity of the irradiation precursor system was 5500a.u., and the fluorescence intensity of the system after 2min of irradiation was 2400a.u., so that 9-anthracene- β -propionic acid and fullerene C were contained60The fluorescence intensity of the detection system is obviously reduced after illumination, the reduction rate of the fluorescence intensity is 56 percent, and the response time is fast; meanwhile, the fluorescence signal intensity of the molecules is not obviously reduced along with the extension of the illumination time.
Description of the drawings: the fluorescence signal intensity in FIG. 8 is in the order of 9-anthracene-beta-propionic acid + C from top to bottom60(not illuminated), 9, 10-diphenylanthracene + C60(2 min of illumination), 9, 10-diphenylanthracene + C60(illumination for 4min), 9, 10-diphenylanthracene + C60(illumination for 6min), 9, 10-diphenylanthracene + C60(illumination for 8min), 9, 10-diphenylanthracene + C60(illumination for 10min), 9, 10-diphenylanthracene + C60(12 min of light irradiation) under the same conditions.
Example 3
Sc based on embedded fullerene3N@C80The method for detecting oxygen by adopting the fluorescent molecular probe 9, 10-diphenylanthracene comprises the following steps:
(1) embedding fullerene Sc3N@C80Dissolving in carbon disulfide to prepare the embedded fullerene Sc3N@C80At a concentration of 10-31, mol/L solution;
(2) dissolving fluorescent molecule 9, 10-diphenyl anthracene in carbon disulfide to prepare 9, 10-diphenyl anthraceneAnthracene concentration of 10- 4mol/L solution 2;
(3) adding the solution 1 into the solution 2 according to the volume ratio of the solution 1 to the solution 2 of 1:1 to obtain the solution containing the embedded fullerene Sc3N@C80The fluorescent molecule solution of (1);
(4) and (3) constructing 7 detection systems according to the step (4) in the embodiment 1, and simultaneously drawing a standard curve graph with the abscissa as the oxygen concentration and the ordinate as the signal intensity according to the step (5) in the embodiment 1 to further obtain a linear function of the fluorescence intensity and the oxygen concentration, so that the concentration of the oxygen in the sample to be detected can be obtained according to the intensity value of the detected fluorescence signal of the sample to be detected.
And (3) verifying the detection result: preparing a new closed container with the volume of 4ml, vacuumizing the detection system which is the same as that in the step (4), introducing 0.74ml of pure oxygen, and after illumination is carried out for 2min, detecting that the fluorescence intensity of the system is 300 a.u.; meanwhile, the function obtained according to the step (4) and the oxygen concentration of 0.74/4-0.185 ml/cm3The theoretical fluorescence intensity was found to be 350a.u., which is within the tolerance of the error, and was different from the detected fluorescence intensity of 300a.u., by only 50a.u. Therefore, the detection method can accurately detect the oxygen content of lower concentration.
As can be seen from FIG. 10, the fluorescence intensity before light irradiation was 2750a.u., and the fluorescence intensity after light irradiation for 2min was 300a.u., so that 9-anthracene- β -propionic acid and endofullerene Sc were contained3N@C80The fluorescence intensity of the detection system is obviously reduced after illumination, the reduction rate of the fluorescence intensity is 89%, and the response time is fast; meanwhile, the fluorescence signal intensity of the molecules is not obviously reduced along with the extension of the illumination time.
Description of the drawings: the fluorescence signal intensity in FIG. 10 is, from top to bottom, 9, 10-diphenylanthracene + Sc3N@C60(not illuminated), 9, 10-diphenylanthracene + Sc3N@C60(2 min of illumination), 9, 10-diphenylanthracene + Sc3N@C60(illumination for 4min), 9, 10-diphenylanthracene + Sc3N@C60(illumination for 6min), 9, 10-diphenylanthracene + Sc3N@C60(illumination for 8min), 9, 10-diphenylanthracene + Sc3N@C60(illumination for 10min), 9, 10-diBenzanthracene + Sc3N@C60(12 min of light irradiation) under the same conditions.
The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
Claims (12)
1. A method for detecting oxygen by adopting a fluorescent molecular probe comprises the following steps:
(1) dissolving fullerene and/or fullerene derivatives in halogenated benzene, alkylbenzene or carbon disulfide to prepare a solution containing the fullerene and/or the fullerene derivatives, namely a solution 1;
(2) dissolving the fluorescent molecules in halogenated benzene, alkylbenzene or carbon disulfide to prepare a solution containing the fluorescent molecules, namely a solution 2;
(3) mixing the solution 1 and the solution 2 to obtain a fluorescent molecule solution containing fullerene and/or fullerene derivatives;
(4) vacuumizing the mixed solution obtained in the step (3) to reduce the oxygen concentration to 0ml/cm3Obtaining a detection system;
(5) under the illumination condition, establishing a linear relation between the fluorescence signal intensity and the oxygen content in a detection system;
(6) introducing a sample to be detected into the detection system same as the step (4), measuring the intensity of a fluorescence signal in the detection system under the same illumination condition as the step (5), and further calculating to obtain the oxygen content in the sample to be detected;
wherein the fullerene is an endohedral fullerene, the endohedral fullerene comprising N @ C60、La@C72、Sc2@C74、Sc2@C76、La2@C80、Sc3N@C80、Tm@C82、Gd@C82、Sc2C2@C84At least one of;
the fullerene derivative comprises at least one of fullerene oxide and fullerene addition product;
the concentration of fullerene and/or fullerene derivative in the solution 1 is 10-6mol/L~10-2mol/L, the concentration of the fluorescent molecules in the solution 2 is 10-6mol/L~10-2mol/L。
2. The method of claim 1, wherein: the fullerene oxide is C60O、C70O, and the fullerene addition product is at least one of fullerene 1, 3-dipolar cycloaddition products and fullerene binger addition products.
3. The method of claim 1, wherein: the fluorescent molecule is an organic molecule with a conjugated structure and a biphenyl structure.
4. The method of claim 3, wherein: the fluorescent molecule is one of 9, 10-diphenylanthracene and 9-anthracene-beta-propionic acid.
5. The method of claim 1, wherein: the halogenated benzene is chlorobenzene, and the alkylbenzene is toluene.
6. The method according to claim 1, wherein the volume ratio of solution 1 to solution 2 is = 1: 3 ~ 3: 1 when solution 1 and solution 2 are mixed in step (3).
7. The method of claim 1, wherein: and (4) in the step (6), the sample to be detected is a gaseous sample or a liquid sample.
8. The method of claim 1, wherein: the illumination conditions include xenon lamp illumination, halogen lamp illumination, and mercury lamp illumination.
9. The method of claim 8, wherein: the lighting conditions include xenon lamp lighting.
10. The method of claim 9, wherein: the xenon lamp irradiation is xenon lamp white light source irradiation.
11. The method according to claim 1 or 8, wherein the intensity of light is 1mW ~ 30mW, and the duration of light is 1min ~ 60 min.
12. The method of claim 1, wherein: the specific steps for establishing the linear relation between the fluorescence signal intensity and the oxygen content in the detection system in the step (5) are as follows: in the detection system, a standard curve graph with the abscissa as the oxygen concentration and the ordinate as the fluorescence signal intensity is drawn according to a plurality of oxygen concentrations and the fluorescence intensity corresponding to each oxygen concentration, and a linear range is determined.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710419871.0A CN107340278B (en) | 2017-06-06 | 2017-06-06 | Method for detecting oxygen by adopting fluorescent molecular probe based on fullerene or fullerene derivative |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710419871.0A CN107340278B (en) | 2017-06-06 | 2017-06-06 | Method for detecting oxygen by adopting fluorescent molecular probe based on fullerene or fullerene derivative |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107340278A CN107340278A (en) | 2017-11-10 |
| CN107340278B true CN107340278B (en) | 2020-01-21 |
Family
ID=60220288
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710419871.0A Active CN107340278B (en) | 2017-06-06 | 2017-06-06 | Method for detecting oxygen by adopting fluorescent molecular probe based on fullerene or fullerene derivative |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107340278B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104086596A (en) * | 2014-05-08 | 2014-10-08 | 南京邮电大学 | Phosphorescent iridium complex, and preparation method and application thereof |
| CN104479670A (en) * | 2014-11-21 | 2015-04-01 | 天津理工大学 | Xanthene fluorescent dye molecular fluorescent probe as well as preparation method and application thereof |
| JP2017003333A (en) * | 2015-06-05 | 2017-01-05 | 国立大学法人大阪大学 | Colorimetric agent for halogen ion and singlet oxygen, and singlet oxygen detecting agent |
| CN106589400A (en) * | 2016-12-22 | 2017-04-26 | 河南理工大学 | Application of mercapto silver cluster coordination polymer in detection of oxygen |
| CN106749421A (en) * | 2016-11-21 | 2017-05-31 | 北京师范大学 | The preparation and its application of highly sensitive singlet oxygen phosphorescence probe |
-
2017
- 2017-06-06 CN CN201710419871.0A patent/CN107340278B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104086596A (en) * | 2014-05-08 | 2014-10-08 | 南京邮电大学 | Phosphorescent iridium complex, and preparation method and application thereof |
| CN104479670A (en) * | 2014-11-21 | 2015-04-01 | 天津理工大学 | Xanthene fluorescent dye molecular fluorescent probe as well as preparation method and application thereof |
| JP2017003333A (en) * | 2015-06-05 | 2017-01-05 | 国立大学法人大阪大学 | Colorimetric agent for halogen ion and singlet oxygen, and singlet oxygen detecting agent |
| CN106749421A (en) * | 2016-11-21 | 2017-05-31 | 北京师范大学 | The preparation and its application of highly sensitive singlet oxygen phosphorescence probe |
| CN106589400A (en) * | 2016-12-22 | 2017-04-26 | 河南理工大学 | Application of mercapto silver cluster coordination polymer in detection of oxygen |
Non-Patent Citations (3)
| Title |
|---|
| Efficient two-photon fluorescent probe for monitoring mitochondrial singlet oxygen in tissues during photodynamic therapy;Hong-wen Liu等;《ROYAL SOCIETY OF CHEMISTRY》;20160921;第1-4页 * |
| 富勒烯C60生物学研究进展及其物理化学基础;严庆丰等;《生命科学研究与应用》;19960531;第231-233页 * |
| 蒽环类物质检测单线态氧的研究新进展;尹红菊等;《曲靖师范学院学报》;20160531;第27-32页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107340278A (en) | 2017-11-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ji et al. | LED‐Illuminated NMR spectroscopy: a practical tool for mechanistic studies of photochemical reactions | |
| Farokhi et al. | Kinetic, mechanistic and spectral studies for the oxidation of sulfanilic acid by alkaline hexacyanoferrate (III) | |
| Ruan et al. | A relay strategy for the mercury (II) chemodosimeter with ultra-sensitivity as test strips | |
| Zhu et al. | Small-molecule fluorescent probes for the detection of carbon dioxide | |
| Zhao et al. | A fluorescence-switchable carbon dot for the reversible turn-on sensing of molecular oxygen | |
| CN113603701B (en) | Colorimetric/fluorescent probe for detecting hypochlorite ions and preparation method and application thereof | |
| Hou et al. | A colorimetric and ratiometric fluorescent probe for sulfite based on an intramolecular cleavage mechanism | |
| Bryant et al. | The use of fluorescent probes for the detection of under-film corrosion | |
| Wu et al. | A novel “on-off-on” acylhydrazone-based fluorescent chemosensor for ultrasensitive detection of Pd2+ | |
| Yang et al. | pH-sensitive fluorescent salicylaldehyde derivative for selective imaging of hydrogen sulfide in living cells | |
| Chang et al. | Steam‐sterilizable, fluorescence lifetime‐based sensing film for dissolved carbon dioxide | |
| Luo et al. | A near-infrared ratiometric fluorescent probe for hydrazine and its application for gaseous sensing and cell imaging | |
| CN107340278B (en) | Method for detecting oxygen by adopting fluorescent molecular probe based on fullerene or fullerene derivative | |
| CN113607709B (en) | Application of fluorescent carbon quantum dot in detection of naphthenic acid and detection method of naphthenic acid | |
| Madani et al. | In situ reaction monitoring in photocatalytic organic synthesis | |
| Wan et al. | A BODIPY‐Based Far‐Red‐Absorbing Fluorescent Probe for Hypochlorous Acid Imaging | |
| CN111116418A (en) | Aggregation-induced luminescent material and preparation method and application thereof | |
| Liu et al. | A new highly selective fluorescent sensor based on a novel fluorophore for cyanide and its applications in bioimaging | |
| Cheng et al. | Reaction‐based probe for hydrogen sulfite: dual‐channel and good ratiometric response | |
| Dahlén et al. | Microwave‐Assisted Generation of Lanthanide (ii) Halides in THF and Simple Quantitative Determination | |
| Qiu et al. | A dual-site colorimetric fluorescent probe for rapid detection of hydrazine/hypochlorite and its application in two-photon fluorescent bioimaging | |
| CN113512039B (en) | Cu detection based on complexation+Fluorescent probe and application thereof | |
| US10746656B2 (en) | CO2 quantitative fluorescent sensing material, preparation method and application thereof | |
| Wu et al. | Chemiluminescence determination of sulphite using a cyclometalated iridium complex as chemiluminescence reagent | |
| CN106431986B (en) | A kind of fluorescence probe for detecting hydrazine and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20180426 Address after: 100064, room 5, 523, room 5, Haidian District trail Road, Beijing Applicant after: BEIJING FUNAKANG BIOTECHNOLOGY CO., LTD. Address before: 100064, room 5, 523, room 5, Haidian District trail Road, Beijing Applicant before: BEIJING FUNAKANG BIOTECHNOLOGY CO., LTD. Applicant before: Institute of Chemistry, Chinese Academy of Sciences |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20201113 Address after: No.9 Wanze Avenue, Hongshan Economic Development Zone, Chifeng City, Inner Mongolia Autonomous Region 024000 Chifeng Funakang Biotechnology Co., Ltd Patentee after: Chifeng Funakang Biotechnology Co.,Ltd. Patentee after: Beijing Fullcan Biotechnology Co.,Ltd. Address before: 100064 room, room 5, No. 5, Haidian District Road, Beijing, China Patentee before: Beijing Fullcan Biotechnology Co.,Ltd. |