Ratiometric probe based on internal filtering effect and construction method and application thereof
阅读说明：本技术 一种基于内过滤效应的比率探针及其构建方法和应用 (Ratiometric probe based on internal filtering effect and construction method and application thereof ) 是由 汪晶 邱昱 李大权 温丝雨 胡军 于 2019-08-28 设计创作，主要内容包括：本发明涉及荧光检测技术领域,尤其涉及一种基于内过滤效应的比率探针及其构建方法和应用。所述比率探针包括n种发光碳点和m种吸光能量受体；所述n种发光碳点依次为A<Sub>1</Sub>发光碳点……A<Sub>n</Sub>发光碳点,且n≥1；所述n种发光碳点的形成x种不重合的荧光发射峰,x≥2,且发光碳点具有氰反应惰性；所述m种吸光能量受体依次为B<Sub>1</Sub>吸光能量受体……B<Sub>m</Sub>吸光能量受体,且m≥1；所述m种吸光能量受体形成y种荧光吸收峰,y种荧光吸收峰与v种荧光发射峰存在重合,且1≤v≤(x-1)；所述吸光能量受体具有氰反应活性。本发明比率探针通过多化学信号检测的方式提高了检测精度和灵敏度,并且具有低毒甚至无毒、检测范围大、构建简便高效等优点。(The invention relates to the technical field of fluorescence detection, in particular to a ratiometric probe based on an internal filtering effect and a construction method and application thereof. The ratiometric probe comprises n luminescent carbon dots and m light absorption energy receptors; the n kinds of luminescent carbon dots are A in sequence 1 Luminescent carbon dot … … A n Luminescent carbon dots, and n is more than or equal to 1; the n luminescent carbon points form x non-coincident fluorescence emission peaks, x is more than or equal to 2, and the luminescent carbon points have cyanogen reaction inertia; the m light absorption energy receptors are sequentially B 1 Light absorbing energy acceptor … … B m A light absorbing energy receptor, and m is more than or equal to 1; the m light absorption energy receptors form y fluorescence absorption peaks, the y fluorescence absorption peaks and the v fluorescence emission peaks are overlapped, and v is more than or equal to 1 and less than or equal to (x-1); the light absorbing energy acceptor is cyanogen reactive. The ratiometric probe improves the detection precision and sensitivity by a multi-chemical signal detection mode, and has the advantages of low toxicity, no toxicity, large detection range, simple and convenient construction, high efficiency and the like.)
1. A ratiometric probe based on an internal filtering effect,
the ratiometric probe comprises n luminescent carbon dots and m light absorption energy receptors;
the n kinds of luminescent carbon dots are A in sequence 1Luminescent carbon dot … … A nThe light-emitting carbon dots are arranged on the substrate,and n is more than or equal to 1;
the n luminescent carbon points form x non-coincident fluorescence emission peaks, x is more than or equal to 2, and the luminescent carbon points have cyanogen reaction inertia;
the m light absorption energy receptors are sequentially B 1Light absorbing energy acceptor … … B mA light absorbing energy receptor, and m is more than or equal to 1;
the m light absorption energy receptors form y fluorescence absorption peaks, the y fluorescence absorption peaks and the v fluorescence emission peaks are overlapped, and v is more than or equal to 1 and less than or equal to (x-1);
the light absorbing energy acceptor is cyanogen reactive.
2. An internal filtering effect based ratiometric probe of claim 1,
the ratiometric probe consists of two luminescent carbon dots and a light absorption energy receptor;
the luminescent carbon dots are respectively A 1Luminescent carbon dots and A 2Luminescent carbon dots wherein A 1The luminescent carbon dots are citric acid carbon dots A 2The luminescent carbon dots are p-phenylenediamine carbon dots;
the light-absorbing energy acceptor is B 1Light-absorbing energy acceptor, B 1The light absorbing energy receptors are silver nanoparticles.
3. An internal filtering effect based ratiometric probe of claim 2,
the citric acid carbon dots are used as blue fluorescent carbon dots to emit blue fluorescence to form a blue fluorescence emission peak, and the peak top wavelength of the blue fluorescence emission peak is 425-435 nm;
the p-phenylenediamine carbon dots are used as red fluorescent carbon dots to emit red fluorescence to form a red fluorescence emission peak, and the peak top wavelength of the red fluorescence emission peak is 583-602 nm;
the silver nanoparticles form a fluorescence absorption peak, and the peak top wavelength of the fluorescence absorption peak is 385-420 nm;
the ratio of the initial intensity of the blue fluorescence to the initial intensity of the red fluorescence is (31-40): 1.
4. a method of constructing an internal filtering effect-based ratiometric probe of claim 1,
the construction method comprises the following steps:
adding n luminescent carbon dots into water, uniformly dispersing to form a dispersion liquid, adding m light absorption energy receptors into the dispersion liquid, and uniformly mixing to obtain a solution, namely the ratiometric probe based on the internal filtration effect.
5. The method for constructing a ratiometric probe based on the internal filtering effect of claim 4,
two luminescent carbon dots are added, namely a citric acid carbon dot and a p-phenylenediamine carbon dot;
one light absorption energy receptor is added, and is silver nanoparticles;
wherein: based on the mass of the carbon dots of the citric acid, the addition amount of the carbon dots of the p-phenylenediamine is 13-24 mg/mg, the amount of water is 0.5-1.5L/mg, and the amount of the silver nanoparticles is 35-59.2 mg/mg.
6. The method for constructing the ratiometric probe based on the internal filtration effect of claim 5, wherein the silver nanoparticles are prepared by the following steps:
adding silver nitrate into water, heating to boil, and adding sodium citrate and sodium borohydride for reduction reaction to obtain the silver nanoparticles.
7. Use of an internal filtering effect-based ratiometric probe according to claim 1, 2 or 3,
the ratiometric probe based on the internal filtering effect is used for cyanide ion detection;
the detection comprises qualitative detection and quantitative detection.
8. Use of an internal filtering effect based ratiometric probe according to claim 7,
the internal filtering effect-based ratiometric probe for qualitatively detecting cyanide ions comprises the following steps:
and performing fluorescence spectrum detection on the ratio probe based on the internal filtration effect as a blank sample to obtain a fluorescence intensity ratio, uniformly mixing the ratio probe based on the internal filtration effect, a sodium carbonate-sodium bicarbonate buffer solution and the solution to be detected in a certain proportion to form a mixed solution, sampling the mixed solution to perform fluorescence spectrum detection, comparing the mixed solution with the fluorescence intensity ratio of the blank sample, and if a significant difference exists, determining that the solution to be detected contains cyanide ions to complete qualitative detection.
9. Use of an internal filtering effect based ratiometric probe according to claim 7,
the internal filtering effect-based ratio probe for quantitatively detecting the cyanide ions comprises the following steps:
1) selecting the same ratiometric probes based on the internal filtration effect, respectively preparing cyanogen ion standard solutions with different concentrations as standard solutions to be detected, respectively and uniformly mixing the ratiometric probes based on the internal filtration effect, a sodium carbonate-sodium bicarbonate buffer solution and each standard solution to be detected according to a certain proportion to form a standard mixed solution, then sampling the standard mixed solution for fluorescence spectrum detection, respectively recording the obtained results and drawing a standard curve of the ratio of the hydrogen ion concentration to the fluorescence intensity;
2) selecting a ratiometric probe based on the internal filtration effect which is the same as the ratiometric probe based on the internal filtration effect in the step 1), uniformly mixing the ratiometric probe based on the internal filtration effect, the sodium carbonate-sodium bicarbonate buffer solution and the liquid to be detected according to a certain proportion to form a mixed solution, then sampling the mixed solution for fluorescence spectrum detection, and substituting the obtained fluorescence intensity ratio into the standard curve obtained in the step 1), so that the concentration of the cyanide ions in the liquid to be detected can be obtained, and the quantitative detection is completed.
Technical Field
The invention relates to the technical field of fluorescence detection, in particular to a ratiometric probe based on an internal filtering effect and a construction method and application thereof.
Background
Cyanide is a deadly toxic anion and, in addition to being produced by biological processes of plants, its use in industry also increases the risk of contamination of water supplies. Cyanogen ions in the environment enter a human body, and can strongly inhibit the catalytic activity of cytochrome c oxidase, so that the respiratory system and the nervous system are disturbed and even die. Therefore, there is a need to develop an effective and feasible method for detecting cyanide ions in the environment. The method for detecting the cyanide ions comprises a titration method, a photoelectrochemical method, a liquid chromatography-mass spectrometry method and an ion chromatography method, the pretreatment of the sample is complex, and the test time is long.
The fluorescence analysis method for detecting ions has the advantages of rapid response, high sensitivity, low cost and the like, and is commonly used for rapid real-time detection. Most of the existing fluorescent probes for detecting cyanide ions are single chemical signal detection and are easily influenced by changes of excitation light intensity, probe concentration, instrument factors and the like to achieve result accuracy. The ratio detection method with self-calibration function developed in recent years can improve the detection sensitivity and realize visual identification of analytes through hue difference. Organic dye, heavy metal quantum dots and carbon dots are often used as fluorophores to construct probes, and compared with the former two, the carbon dots have the advantages of low toxicity, good water solubility, controllable ligand modification groups and strong light stability. The silver nanoparticles have the characteristics of easy chemical modification, high extinction coefficient and size-dependent optical characteristic change, and can be used as an energy acceptor to quench donor fluorescence. Designing fluorescent probes by a combination of fluorophores and quenchers is the most attractive strategy to address the single component weaknesses and to optimize probe performance.
For example, the application publication number of the invention patent application of the nano silver fluorescent probe for detecting cyanide ions and the preparation method thereof, which is disclosed by the chinese patent office at 8.8.2012, is CN102627966A, the nano silver fluorescent probe with the sequence 5'-AAAAAAAACCCCCCCCCTTTTTTTT-3' is prepared by using oligonucleotide as a template and reacting under the action of a reducing agent sodium borohydride, the preparation period is long, the toxicity resistance (namely, the impurity ion interference resistance) is limited, and only qualitative detection but no quantitative detection can be performed.
Also, for example, the application of the invention patent application of the application method of the ratiometric cyanide ion fluorescent probe molecule disclosed by the chinese patent office at 2013, 6/5/No. CN103134787A, the application publication is CN103134787A, the cyanide ion is detected by using 1, 2, 2, 3-tetramethyl-4, 5-benzindoline, and the fluorescence color is changed to perform qualitative detection mainly by shifting or reconstructing the fluorescence emission peak, so that the method is difficult to be effectively used for quantitative detection, and the chemical signal is single and is easily influenced by the excitation light intensity.
Disclosure of Invention
The invention provides a ratiometric probe based on an internal filtering effect, a construction method and application thereof, aiming at solving the problems that the existing fluorescent probe for detecting cyanide ions has single chemical signal and is easily interfered by external environment, the existing fluorescent probe has weak toxicity resistance, is very easily interfered when impurity ions exist in a liquid to be detected, and part of the fluorescent probe can be interfered even by chloride ions, so that the using condition is severe, and the popularization and application are difficult. The purpose is as follows: firstly, realizing multi-chemical signal detection and improving detection precision; the antitoxicity of the ratiometric probe is improved, the interference of impurity ions to the ratiometric probe is reduced, and the ratiometric probe has higher stability, and further has wide universality and higher use convenience; thirdly, the toxicity of the ratiometric probe is reduced, the water solubility of the ratiometric probe is improved, and the detection application of an actual sample is facilitated; fourthly, the combination of a fluorophore and a quencher is adopted to realize internal filtration, and the detection range of the ratiometric probe is enlarged; fifthly, the construction method of the ratiometric probe is simplified, and the preparation efficiency is improved; sixthly, the qualitative detection and the quantitative detection of the cyanide ions can be rapidly realized.
In order to achieve the purpose, the invention adopts the following technical scheme.
A ratiometric probe based on an internal filtering effect,
the ratiometric probe comprises n luminescent carbon dots and m light absorption energy receptors;
the n kinds of luminescent carbon dots are A in sequence 1Luminescent carbon dot … … A nLuminescent carbon dots, and n is more than or equal to 1;
the n luminescent carbon points form x non-coincident fluorescence emission peaks, x is more than or equal to 2, and the luminescent carbon points have cyanogen reaction inertia;
the m light absorption energy receptors are sequentially B 1Light absorbing energy acceptor … … B mLight absorbing energy receiverA body, and m is more than or equal to 1;
the m light absorption energy receptors form y fluorescence absorption peaks, the y fluorescence absorption peaks and the v fluorescence emission peaks are overlapped, and v is more than or equal to 1 and less than or equal to (x-1);
the light absorbing energy acceptor is cyanogen reactive.
Wherein the luminescent carbon dots (i.e., fluorescent carbon dots) are used as fluorophores, and the light absorption energy receptor is used as a quencher, and the two are combined to realize internal filtration, so that the internal filtration effect is generated. And the y fluorescence absorption peaks partially or completely coincide with the fluorescence emission peak in the v, wherein the coincidence concept is the obvious overlap of peak areas, when the spectra coincide, the emitted fluorescence can be absorbed by a light absorption energy receptor, so that the fluorescence intensity is reduced, and the total intensity of the fluorescence emission peak which is not coincident with the fluorescence absorption peak in the luminescent carbon point is set as I AThe total intensity of a fluorescence emission peak coincident with a fluorescence absorption peak in the luminescent carbon dots is I BThe ratio of fluorescence intensities (I) before addition and reaction of the quencher (absorbing energy acceptor) B/I A) 0＝a 0Fluorescence intensity ratio (I) after addition and reaction of a quencher (absorbing energy acceptor) B/I A) 1＝a 1，a 1＜a 0. Therefore, in the actual use process, the ratio of the actual fluorescence intensity of the ratiometric probe based on the internal filtering effect is a 1When the device is used for detecting liquid cyanide ions to be detected, the cyanide ions can etch a light absorption energy receptor with cyanide reaction activity, and the luminescent carbon dots have cyanide reaction inertia and do not react with the cyanide ions, so that the light absorption capability of the luminescent carbon dots is reduced while the fluorescence emission capability of the luminescent carbon dots is maintained, and a is ensured 1Gradually trend to a 0The change of the fluorescence intensity ratio can be rapidly detected through an instrument, high-efficiency qualitative analysis is realized, certain formula relevance exists between the concentration of the cyanide ions to be detected and the fluorescence intensity ratio in the process, a corresponding relevance formula can be derived by drawing a standard curve, and then rapid and accurate quantitative detection can be realized.
As a preference, the first and second liquid crystal compositions are,
the ratiometric probe consists of two luminescent carbon dots and a light absorption energy receptor;
the luminescent carbon dots are respectively A 1Luminescent carbon dots and A 2Luminescent carbon dots wherein A 1The luminescent carbon dots are citric acid carbon dots A 2The luminescent carbon dots are p-phenylenediamine carbon dots;
the light-absorbing energy acceptor is B 1Light-absorbing energy acceptor, B 1The light absorbing energy receptors are silver nanoparticles.
Two different fluorescence emission peaks formed by the citric acid carbon dots and the p-phenylenediamine carbon dots are matched with the nano silver particles as light absorption energy receptors, the nano silver particles mainly absorb light emitted by the citric acid carbon dots, the fluorescence absorption peaks of the nano silver particles are overlapped with the fluorescence emission peaks of the citric acid carbon dots in a large area and are not overlapped with the fluorescence emission peaks of the p-phenylenediamine carbon dots, and a good internal filtering effect can be generated.
As a preference, the first and second liquid crystal compositions are,
the citric acid carbon dots are used as blue fluorescent carbon dots to emit blue fluorescence to form a blue fluorescence emission peak, and the peak top wavelength of the blue fluorescence emission peak is 425-435 nm;
the p-phenylenediamine carbon dots are used as red fluorescent carbon dots to emit red fluorescence to form a red fluorescence emission peak, and the peak top wavelength of the red fluorescence emission peak is 583-602 nm;
the silver nanoparticles form a fluorescence absorption peak, and the peak top wavelength of the fluorescence absorption peak is 385-420 nm;
the ratio of the initial intensity of the blue fluorescence to the initial intensity of the red fluorescence is (31-40): 1.
the ratio of the initial intensity of the blue fluorescence to the initial intensity of the red fluorescence is (I) B/I A) 0＝a 0. The large-area overlapping of the fluorescence emission peak and the fluorescence absorption peak of the citric acid carbon point, the silver nanoparticles can generate the obvious absorption effect on the fluorescence emitted by the citric acid carbon point, so that the generated actual fluorescence intensity ratio (I) B/I A) 1＝a 1Compared with the initial fluorescence intensity ratio (I) B/I A) 0＝a 0There is a significant difference, the larger, the more significant the difference, the later qualitative detectionThe higher the detection sensitivity and the detection accuracy in quantitative detection can be generated in the detection process. At the same time, a 1And a 0Also depends on the initial cardinality value a 0Is controlled by the size of (a) and thus 0The above (31-40): 1, while ensuring that the fluorescence color of the probe corresponding to the base number is blue, and a 1And a 0Has a large difference value of a 1The corresponding fluorescent color of the probe is red, so that the detection precision and visual detection of the ratiometric probe are ensured. Preferably, the peak top wavelength of the blue fluorescence emission peak is 429 to 433nm, the peak top wavelength of the red fluorescence emission peak is 584 to 590nm, and the initial fluorescence intensity ratio a 0Is (35-38): 1, which is optimally: the peak top wavelength of the blue fluorescence emission peak is 431nm, the peak top wavelength of the red fluorescence emission peak is 584nm, and the initial fluorescence intensity ratio a 0Is 38: 1.
a construction method of a ratiometric probe based on an internal filtering effect,
the construction method comprises the following steps:
adding n luminescent carbon dots into water, uniformly dispersing to form a dispersion liquid, adding m light absorption energy receptors into the dispersion liquid, and uniformly mixing to obtain a solution, namely the ratiometric probe based on the internal filtration effect.
The construction method is simple and efficient, the ratiometric probe based on the internal filtration effect can be prepared by simply dispersing a plurality of luminescent carbon dots to prepare a dispersion liquid, adding a plurality of light absorption energy receptors to serve as quenchers after forming the ratiometric fluorophore, and the prepared ratiometric probe is a liquid preparation and is easier to uniformly disperse in a liquid to be detected during use.
As a preference, the first and second liquid crystal compositions are,
two luminescent carbon dots are added, namely a citric acid carbon dot and a p-phenylenediamine carbon dot;
one light absorption energy receptor is added, and is silver nanoparticles;
wherein: based on the mass of the carbon dots of the citric acid, the addition amount of the carbon dots of the p-phenylenediamine is 13-24 mg/mg, the amount of water is 0.5-1.5L/mg, and the amount of the silver nanoparticles is 35-59.2 mg/mg.
The citric acid carbon dots and the p-phenylenediamine carbon dots are common and conventional carbon dot materials, and have the advantages of easy acquisition, easy preparation and the like. The initial fluorescence intensity ratio can be conveniently controlled by controlling the dosage of the two luminescent carbon dots, and the actual fluorescence intensity ratio can be effectively regulated and controlled by controlling the addition of the silver nanoparticles.
As a preference, the first and second liquid crystal compositions are,
the silver nanoparticles are prepared by the following method:
adding silver nitrate into water, heating to boil, and adding sodium citrate and sodium borohydride for reduction reaction to obtain the silver nanoparticles.
The half-height peak width of the fluorescence absorption peak of the conventional silver nanoparticle is large, so that the fluorescence absorption peak is easily overlapped with two fluorescence emission peaks, and as exemplified by the red fluorescence emission peak and the blue fluorescence emission peak, theoretically, the fluorescence absorption peak of the silver nanoparticle needs to be controlled to be overlapped with the blue fluorescence absorption peak as much as possible and not overlapped with the red fluorescence absorption peak as much as possible, so that the initial fluorescence intensity ratio and the actual fluorescence intensity ratio can be controlled to generate a larger difference value, the precision and the visualization capability of quantitative detection are improved, and therefore, certain defects exist in the using process of the conventional silver nanoparticle. And through the preparation method of the double reducing agents, silver nitrate is added into water which is vigorously stirred or vigorously oscillated, sodium citrate and sodium borohydride double reducing agents are added after the mixture is heated to be boiled to carry out reduction reaction, and the prepared silver nanoparticles have the characteristics of small half-height peak width of a fluorescence absorption peak, maximum absorption value corresponding wavelength within the range of 385-420 wavelengths and the like, can always ensure that the overlap area of the fluorescence absorption peak and a blue fluorescence emission peak of a carbon dot of citric acid is large, can control the fluorescence absorption peak not to overlap with a red fluorescence emission peak of a carbon dot of p-phenylenediamine, can generate good internal filtering effect, and are very beneficial to realizing ratio detection and improving detection precision.
An application of ratiometric probes based on the internal filtering effect,
the ratiometric probe based on the internal filtering effect is used for cyanide ion detection;
the detection comprises qualitative detection and quantitative detection.
The ratiometric probe based on the internal filtering effect has the advantages of high sensitivity, high accuracy, high antitoxic capability and the like when being used for detecting cyanide ions, and can quickly carry out high-sensitivity qualitative detection and high-precision quantitative detection.
As a preference, the first and second liquid crystal compositions are,
the internal filtering effect-based ratiometric probe for qualitatively detecting cyanide ions comprises the following steps:
and performing fluorescence spectrum detection on the ratio probe based on the internal filtration effect as a blank sample to obtain a fluorescence intensity ratio, uniformly mixing the ratio probe based on the internal filtration effect, a sodium carbonate-sodium bicarbonate buffer solution and the solution to be detected in a certain proportion to form a mixed solution, sampling the mixed solution to perform fluorescence spectrum detection, comparing the mixed solution with the fluorescence intensity ratio of the blank sample, and if a significant difference exists, determining that the solution to be detected contains cyanide ions to complete qualitative detection.
The detection method is simple and rapid, and qualitative detection can be rapidly carried out by comparing the difference of the fluorescence intensity ratio.
As a preference, the first and second liquid crystal compositions are,
the internal filtering effect-based ratio probe for quantitatively detecting the cyanide ions comprises the following steps:
1) selecting the same ratiometric probes based on the internal filtration effect, respectively preparing cyanogen ion standard solutions with different concentrations as standard solutions to be detected, respectively and uniformly mixing the ratiometric probes based on the internal filtration effect, a sodium carbonate-sodium bicarbonate buffer solution and each standard solution to be detected according to a certain proportion to form a standard mixed solution, then sampling the standard mixed solution for fluorescence spectrum detection, respectively recording the obtained results and drawing a standard curve of the ratio of the cyanogen ion concentration to the fluorescence intensity;
2) selecting a ratiometric probe based on the internal filtration effect which is the same as the ratiometric probe based on the internal filtration effect in the step 1), uniformly mixing the ratiometric probe based on the internal filtration effect, the sodium carbonate-sodium bicarbonate buffer solution and the liquid to be detected according to a certain proportion to form a mixed solution, then sampling the mixed solution for fluorescence spectrum detection, and substituting the obtained fluorescence intensity ratio into the standard curve obtained in the step 1), so that the concentration of the cyanide ions in the liquid to be detected can be obtained, and the quantitative detection is completed.
The method comprises the steps of firstly establishing a correlation between cyanide ion concentration and fluorescence intensity ratio by drawing a standard curve, then measuring the fluorescence intensity ratio of the liquid to be detected and bringing the fluorescence intensity ratio into the correlation, and then quickly obtaining the cyanide ion concentration, wherein the method can ensure that the detection result has higher accuracy.
The invention has the beneficial effects that:
1) the ratiometric probe improves the overall detection precision and detection sensitivity in a multi-chemical signal detection mode;
2) the introduction of low antitoxic components can be avoided through the matching of the quencher and the fluorophore, high antitoxic matching is realized, the antitoxic property of the whole ratiometric probe is improved, the interference of impurity ions on the ratiometric probe can be greatly reduced, the ratiometric probe can be suitable under the condition of a huge number of pairs, and the ratiometric probe has wide universality, higher stability and convenience in use;
3) the toxicity of the whole ratiometric probe is reduced by adopting low-toxicity or even non-toxic luminescent carbon spots, so that the low-toxicity or even non-toxic detection is realized;
4) the internal filtering effect is realized through the combination of the fluorophore and the quencher, the difference between the initial fluorescence intensity ratio and the actual fluorescence intensity ratio is controlled, and the detection range of the ratiometric probe can be enlarged;
5) the whole ratiometric probe construction method is simple and efficient, and the purpose of quickly constructing the ratiometric probe can be realized;
6) can realize high-sensitivity qualitative detection and high-precision quantitative detection of cyanide ions.
Drawings
FIG. 1 is a schematic diagram of the construction and detection mechanism of the ratiometric probe based on the internal filtering effect according to the present invention;
FIG. 2 is a fluorescence spectrum of two carbon dots and a UV spectrum of silver nanoparticles in example 1;
FIG. 3 is a fluorescence spectrum of a ratiometric probe constructed in example 1 of the present invention in response to cyanide ions;
FIG. 4 is a standard curve diagram of the ratio of cyanide ion concentration to fluorescence intensity;
FIG. 5 is a graph of the fluorescence ratio responses of ratiometric probes constructed in example 1 of the present invention to different anions.
Detailed Description
The invention is described in further detail below with reference to specific embodiments and the attached drawing figures. Those skilled in the art will be able to implement the invention based on these teachings. Moreover, the embodiments of the present invention described in the following description are generally only some embodiments of the present invention, and not all embodiments. Therefore, all other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present invention without any creative effort shall fall within the protection scope of the present invention.
Unless otherwise specified, the raw materials used in the examples of the present invention are all commercially available or available to those skilled in the art; unless otherwise specified, the methods used in the examples of the present invention are all those known to those skilled in the art.