CN110057801B - Fluorescence ratiometric probe based on aggregation-induced emission property and application of fluorescence ratiometric probe in detection of hydrogen peroxide and glucose - Google Patents
Fluorescence ratiometric probe based on aggregation-induced emission property and application of fluorescence ratiometric probe in detection of hydrogen peroxide and glucose Download PDFInfo
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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- G01N21/64—Fluorescence; Phosphorescence
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Abstract
The invention relates to a fluorescence ratio probe based on aggregation-induced emission property and application thereof in detecting hydrogen peroxide and glucose, belonging to the cross field of chemistry and materials science. The probe consists of gold nanoclusters coated by tetraphenyl ethylene sulfonate and bovine serum albumin. The electrostatic adsorption of the tetraphenyl ethylene sulfonate and the tetraphenyl ethylene sulfonate causes aggregation-induced luminescence, the mixed solution has double emission wavelengths between 410-550nm and 550-850nm respectively, the proportion of the mixed solution and the 550-850nm is different, and the colors presented by the mixed fluorescent probe are different. The aggregation-induced emission of tetraphenylethenesulfonate can be used as a reference fluorescence color according to the change value of the fluorescence ratio (I) of the double emission peaks0‑I)/I0The linear change relationship with the analyte concentration C realizes the detection of hydrogen peroxide and glucose, and the fluorescence ratiometric probe has the advantages of simple, quick and visual detection of hydrogen peroxide and glucose.
Description
Technical Field
The invention relates to a fluorescence ratio probe based on aggregation-induced emission property and application thereof in detecting hydrogen peroxide and glucose, relates to preparation of a nano material and detection of hydrogen peroxide and glucose, and belongs to the cross field of chemistry and materials science.
Background
Hydrogen peroxide (H)2O2) Is an important active oxygen, which is widely applied to industrial and household washing, bleaching and medical disinfection, but the high-concentration hydrogen peroxide residue in the wastewater can cause serious and irreversible oxidative damage to aquatic organisms. On the other hand, hydrogen peroxide, as an important biological signal molecule, plays an important role in a variety of biological processesAnd (4) color. Therefore, hydrogen peroxide detection is of vital importance in both the environmental protection and biomedical research fields.
Glucose is an important substance in the metabolic balance of the human body and is an energy source for maintaining the life activities of the human body. Blood glucose level is one of the indexes of health of a human body, and too high or too low blood glucose level indicates diabetes or hypoglycemia, so that detection of blood glucose concentration is very important in clinical diagnosis.
At present, a plurality of methods for detecting the content of hydrogen peroxide and glucose are available, including an electrochemical method, a fluorescence detection method, a near infrared spectroscopy method and the like, wherein the fluorescence probe detection has the advantages of simplicity, low cost, high sensitivity and the like. For example, document "H2O2It is reported that hydrogen peroxide quenches the luminescence of cadmium telluride-cadmium sulfide quantum dots, glucose oxidase catalyzes glucose to generate gluconic acid and hydrogen peroxide, and the simple and rapid detection of the glucose can be realized by detecting the hydrogen peroxide. As another example, the document "Fluorescent detection of hydrogen peroxide and glucose with polyethylene-immobilized Cu nanocrusters," Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy,2014,118: 315-. In recent years, colorimetric fluorescence technology has attracted much attention due to its high sensitivity and accuracy, and its inherent corrective effect on the environment. The colorimetric fluorescence technology realizes detection based on the change of the fluorescence ratio of two different wavelengths before and after detection, and compared with a single-change fluorescent probe, the ratio of the color change can be clearly distinguished by naked eyes, so that semi-quantitative detection is realized. For example, the document "graphic carbon nitride sheets-based radiometric fluorescent probe for high sensitivity detection of H2O2and glucose,ACS Applied Materials&Interfaces,2016,8(49):33439-33445.”The ratiometric fluorescent probe for detecting hydrogen peroxide and glucose with high sensitivity based on the carbon nitride nanosheet is reported, o-phenylenediamine is oxidized by hydrogen peroxide in the presence of horseradish peroxidase, an oxidation product is accumulated on the carbon nitride nanosheet to quench the fluorescence of the carbon nitride nanosheet, a new emission peak is generated, and high-flux and low-cost detection of glucose can be realized on the basis of hydrogen peroxide generated by catalysis of glucose oxidase. As another example, the document "A dual-emission nanoparticles of gold nanoparticles and carbon dots for quantitative fluorescence detection of reactive oxygen species and glucose, Journal of biological Nanotechnology,2017,13(11): 1425-. Also for example, the document "A novel reduced reactivity fluorescent probe for the determination of H2O2and glucose via etching of silver nanoparticles, analysis, 2019, 144(4): 1153-.
The conventional fluorescent material, such as inorganic quantum dots, metal nanoclusters, carbon dots and the like, can only emit light in a dilute solution, and can generate self-quenching in a high concentration or aggregation state, so that the sensitivity can be reduced, the detection result can be interfered and the like. Aggregation-Induced Emission (AIE) is a phenomenon in which a type of molecule hardly emits light in a solution, and light Emission is enhanced in an aggregated state or a solid state. The aggregation-induced emission probe has good light stability and portability, has high signal/noise ratio, and has great application prospect in the fields of chemical sensing, biological monitoring and the like.
Disclosure of Invention
The invention aims to provide a fluorescence ratio probe based on aggregation-induced emission property and application thereof in detecting hydrogen peroxide and glucose, aiming at the problems that the traditional fluorescence probe is easy to generate self-quenching in rapid detection, the detection result is easy to interfere and the like.
In order to achieve the purpose, the invention adopts the following technical scheme:
a fluorescence ratiometric probe based on aggregation-induced emission properties is composed of tetraphenyl ethylene sulfonate and bovine serum albumin coated gold nanoclusters, wherein the tetraphenyl ethylene sulfonate is used as a reference fluorescence probe, the bovine serum albumin coated gold nanoclusters are used as fluorescence probes, the tetraphenyl ethylene sulfonate and the bovine serum albumin coated gold nanoclusters are mixed to construct the fluorescence ratiometric probe, and the color of a mixed solution presents red, yellow or blue according to different mixing ratios of the tetraphenyl ethylene sulfonate and the bovine serum albumin.
A method for preparing a fluorescence ratio probe based on aggregation-induced emission properties comprises the following specific steps:
step one, preparing a tetraphenyl ethylene sulfonate solution, weighing a certain amount of tetraphenyl ethylene sulfonate, and dissolving the tetraphenyl ethylene sulfonate in water.
And step two, preparing the gold nanoclusters coated by the bovine serum albumin, adding 5mL of chloroauric acid (5-20mM) into 5mL of bovine serum albumin solution (20-50mM), adjusting the pH value to be alkaline, and reacting at 25-60 ℃ for 10-24 hours to obtain the gold nanoclusters (BSA-Au NCs) coated by the bovine serum albumin.
Step three, mixing the tetraphenyl ethylene sulfonate prepared in the step one with the gold nanoclusters prepared in the step two to obtain a probe;
step three, the compounding method comprises the following steps: and (2) mixing the solution (without fluorescence) of tetraphenyl ethylene sulfonate (chemical structural formula is shown as formula 1) prepared in the first step with the gold nanoclusters coated by the bovine serum albumin synthesized in the second step, wherein the gold nanoclusters coated by the bovine serum albumin are positively charged, the tetraphenyl ethylene sulfonate is negatively charged, the gold nanoclusters coated by the tetraphenyl ethylene sulfonate and the bovine serum albumin are subjected to electrostatic adsorption and aggregation induced luminescence, and the fluorescence emission wavelength of the gold nanoclusters is 400-550 nm. The fluorescence emission wavelength of the mixed solution is respectively 400-550nm and 550-850nm, and the ratio type fluorescence probe with dual-wavelength emission is obtained.
Wherein: r1、R2＝Cl、Br、I、CnH2n+1,n＝0，1，2，3……n。
Preferably, in step one, when n is 0, the tetraphenyl ethylene sulfonate is R1、R2H, i.e. tetraphenylethylenesulfonate, is 1, 2-bis [4- (3-sulfopropoxy) phenyl]1, 2-diphenylethylene sodium salt (BSPOTPE), with the preparation concentration of 0.1 mg/mL; the concentration of the gold nanoclusters coated by the bovine serum albumin prepared in the step two is 5mM (in Au in chloroauric acid)3+Concentration meter of ions); in the third step, the volume ratio of the tetraphenyl ethylene sulfonate solution to the bovine serum albumin coated gold nanoclusters is 1: 0.5.
The invention also relates to a method for detecting hydrogen peroxide or glucose by using the probe, which comprises the following steps:
dropwise adding the solution to be detected into the probe solution, reacting for a certain time, and if the color of the probe solution is changed by visual inspection, indicating that the solution to be detected contains hydrogen peroxide or glucose;
the ratio of fluorescence intensity at 680nm and 490nm of the initial probe (i.e., F) was recorded680/F490)I0Respectively adding the known solutions to be detected with different concentrations C into the probe solution, recording the fluorescence intensity ratio I after the color change of the probe solution is stable, and obtaining the change value (I) according to the fluorescence intensity ratio0-I)/I0And the change rule of the concentration C of the solution to be detected to obtain a linear relation equation: (I)0-I)/I0A is the slope of a linear equation and b is the intercept of the linear equation;
(1) fluorescence ratiometric probe detection
Quantitative detection of target analytes: the target analyte can quench the fluorescence of the gold nanocluster coated by the bovine serum albumin with the emission wavelength of 550-850nm, and the fluorescence of the tetraphenyl ethylene sulfonate at 400-550nm is not influenced, so that the tetraphenyl ethylene sulfonate can be used as a fluorescence background reference color to construct a dual-emission ratio type fluorescence probe. Recording fluorescenceRatio of intensities I and I0I and I0The ratio of the fluorescence intensity at an emission wavelength of 680nm to the fluorescence intensity at 490nm at the sample concentration and blank, respectively (F)680/F490) Calculating a fluorescence ratio change value (I)0-I)/I0And the linear relation with the target analyte concentration C, and carrying out quantitative analysis on the sample with unknown concentration by using a linear relation equation.
Visual semi-quantitative detection of target analytes: under 365nm ultraviolet light irradiation, different concentrations of analytes affect the ratio of the fluorescence intensity at the emission wavelength of 680nm to the fluorescence intensity at 490nm, and the solution color shows red-yellow-green change according to the change of the analyte concentration. The response degree of the analyte can be distinguished by naked eyes according to the color change, semi-quantitative detection of the analyte is realized, and the whole process is convenient and accurate.
The ratio type fluorescent probe of the invention detects hydrogen peroxide: mixing hydrogen peroxide solution with fluorescence ratio probe, adding buffer solution (pH 5-8), diluting to constant volume, reacting at 25-50 deg.C for more than 30min, scanning fluorescence emission wavelength of 410-850nm with fluorescence instrument, and recording fluorescence intensity ratio I and I0I and I0The ratio of the fluorescence intensity at an emission wavelength of 680nm to the fluorescence intensity at 490nm at the sample concentration and blank, respectively (F)680/F490) According to the change value (I) of the ratio of fluorescence of the two emission peaks0-I)/I0And establishing a standard curve according to the relation between the concentration C of the hydrogen peroxide solution, and comparing the standard curve to obtain the concentration of the hydrogen peroxide in the sample to be detected. Semi-quantitative analysis by visual colorimetry under 365nm ultraviolet light
The ratio type fluorescent probe of the invention is used for detecting glucose: mixing a glucose solution and a glucose oxidase solution, adding a buffer solution (pH 5-8), adding a fluorescence ratio probe, adding water to a constant volume, uniformly mixing, reacting at 25-50 ℃ for more than 30min, scanning the fluorescence emission wavelength of 850nm and 410-inch by using a fluorescence instrument, and recording fluorescence intensity ratios I and I0I and I0The ratio of the fluorescence intensity at an emission wavelength of 680nm to the fluorescence intensity at 490nm at the sample concentration and blank, respectively (F)680/F490) According to twoChange in ratio of fluorescence of emission Peak (I)0-I)/I0And establishing a standard curve according to the relation between the concentration C of the glucose solution, and comparing the standard curve to obtain the concentration of the glucose in the sample to be detected. Semi-quantitative analysis was performed by visual colorimetry under 365nm ultraviolet light.
The invention has the beneficial effects that:
1. a fluorescence ratio probe is constructed based on the aggregation-induced luminescence characteristic of tetraphenyl ethylene sulfonate caused by the electrostatic adsorption effect of the gold nanocluster coated by the bovine serum albumin and the tetraphenyl ethylene sulfonate and the property that the gold nanocluster coated by the bovine serum albumin emits red fluorescence, wherein the ratio of the fluorescence ratio probe and the gold nanocluster is different, and the mixed solution shows red-yellow-green color change.
2. Hydrogen peroxide or hydrogen peroxide generated by the reaction of glucose and glucose oxidase can cause the red fluorescence quenching of the gold nanoclusters coated by bovine serum albumin without influencing the blue-green fluorescence of tetraphenyl ethylene sulfonate, so that the aggregation-induced emission of tetraphenyl ethylene sulfonate can be used as a reference fluorescence color according to the change value of the dual-emission-peak fluorescence ratio (I)0-I)/I0The linear variation relationship with the analyte concentration C enables the detection of the target analyte. The reference fluorescence color can reduce data errors and the like caused by factors such as substrate, external environment, instrument condition change and the like, and can be used for detecting actual samples. The ratiometric probe has the advantages of rapidness, simplicity and capability of realizing quantitative detection by using a fluorimeter and realizing visual semi-quantitative detection under ultraviolet light, is used for detecting hydrogen peroxide and glucose in a ratio type and a visual mode by taking a tetraphenyl ethylene sulfonate aggregation induced luminescent probe caused by gold nanoclusters coated by bovine serum albumin as a mixed fluorescence ratiometric probe, has a very wide application prospect in the field of rapid detection of hydrogen peroxide and glucose, and is expected to be used for detecting other small molecules.
Drawings
FIG. 1 is a schematic diagram of the principle of detecting hydrogen peroxide and glucose by a ratiometric fluorescent probe;
FIG. 2 shows fluorescence emission spectra of 1, 2-bis [4- (3-sulfopropoxy) phenyl ] -1, 2-stilbene sodium salt (BSPOTPE) and bovine serum albumin coated gold nanoclusters (BSA-AuNCs) at different ratios;
FIG. 3 shows the change of the ratio of hydrogen peroxide to fluorescence (I) at different concentrations0-I)/I0A standard curve of (a);
FIG. 4 shows the change of the ratio of glucose to fluorescence (I) at different concentrations0-I)/I0A standard curve of (a);
FIG. 5 shows the selective detection of glucose and other sugars.
Detailed Description
The invention is further described with reference to the following figures and examples.
Example 1
A fluorescence ratiometric probe based on aggregation-induced emission property is a tetraphenyl ethylene sulfonate and bovine serum albumin coated gold nanocluster composite material, and the color of the composite material is red, yellow or blue according to different mixing ratios of the tetraphenyl ethylene sulfonate and the bovine serum albumin;
a method for preparing a fluorescence ratio probe based on aggregation-induced emission properties comprises the following specific steps:
step one, preparing tetraphenyl ethylene sulfonate solution, weighing a certain amount of 1, 2-bis [4- (3-sulfonic acid propoxy) phenyl ] -1, 2-diphenylethylene sodium salt (BSPOTPE), and dissolving in water, wherein the concentration of the BSPOTPE is 0.1 mg/L.
Step two, preparing gold nanoclusters coated by bovine serum albumin, adding 5mL of chloroauric acid (10mM) into 5mL of bovine serum albumin solution (50mM), adjusting the pH value to 10 by using sodium hydroxide, and reacting at 37 ℃ for 12 hours to obtain the gold nanoclusters coated by the bovine serum albumin (BSA-Au NCs) with the concentration of 5mM (Au in chloroauric acid)3+Concentration meter of ions);
step three, mixing the tetraphenyl ethylene sulfonate solution (0.1mg/L) prepared in the step one with the bovine serum albumin coated gold nanocluster (5mM) prepared in the step two according to any volume ratio to obtain a fluorescence ratio probe, wherein as shown in figure 2, the ratios of the tetraphenyl ethylene sulfonate to the bovine serum albumin coated gold nanocluster are different, and the ratio of the fluorescence intensity of the fluorescence probe at the position of 680nm to the fluorescence intensity of the fluorescence probe at the position of 490nm (F) is obtained680/F490) Different, the color under ultraviolet light shows different colors, and the volume ratio BSPOTPE: when BSA-Au NCs is 1:1, the fluorescent probe is red, and the volume ratio of BSPOTPE: when BSA-Au NCs is 1:0.5, the fluorescent probe is orange, and the volume ratio of BSPOTPE: when BSA-Au NCs is 1:0.1, the fluorescent probe appears green.
Example 2
Detection of hydrogen peroxide: 12.5. mu.L of bovine serum albumin coated gold nanoclusters (5mM, Au in chloroauric acid)3+Concentration of ion) and 25. mu.L of 1, 2-bis [4- (3-sulfopropoxy) phenyl group]-1, 2-stilbene sodium salt (0.1mg/L) mixed to a fluorescent ratiometric probe. 50 mu L H2O2The solutions (1, 2, 3, 4, 5, 6, 7, 8mM) were mixed with the fluorescence ratio probe, 100. mu.L of PBS buffer (10mM, pH7) was added thereto, the volume was adjusted to 500. mu.L, the reaction was carried out at 37 ℃ for 30min, then the fluorescence emission wavelength was scanned with a fluorometer at 410-0I and I0The ratio of the fluorescence intensity at an emission wavelength of 680nm to the fluorescence intensity at 490nm at the sample concentration and blank, respectively (F)680/F490) Obtaining a fluorescence ratio change value (I)0-I)/I0Linear relationship with hydrogen peroxide concentration C (I)0-I)/I00.0322C +0.5347, as shown in fig. 3. With this linear relationship, a quantitative analysis of hydrogen peroxide in an unknown sample can be performed. Semi-quantitative analysis was performed by visual colorimetry under 365nm ultraviolet light.
Example 3
And (3) glucose detection: 12.5. mu.L of bovine serum albumin coated gold nanoclusters (5mM, Au in chloroauric acid)3+Concentration of ion) and 25. mu.L of 1, 2-bis [4- (3-sulfopropoxy) phenyl group]-1, 2-stilbene sodium salt (0.1mg/L) mixed to a fluorescent ratiometric probe. Mixing 100 μ L glucose solution (1, 2, 3, 4, 5, 6, 7, 8, 9mM) and 100 μ L glucose oxidase (0.5mg/L) solution, adding 100 μ L PBS buffer solution (10mM, pH7), adding into fluorescence ratio probe, adding water to a constant volume of 500 μ L, mixing well, reacting at 37 deg.C for 30min, scanning fluorescence emission wavelength of 410-0I and I0Sample concentration and blank, respectivelyRatio of fluorescence intensity at emission wavelength of 680nm to fluorescence intensity at 490nm (F)680/F490) Obtaining a fluorescence ratio change value (I)0-I)/I0Linear relationship with glucose concentration C (I)0-I)/I00.0705C +0.0407, as shown in fig. 4. With this linear relationship, a quantitative analysis of glucose in an unknown sample can be performed. Semi-quantitative analysis was performed by visual colorimetry under 365nm ultraviolet light.
Example 4
Detection of glucose in serum: 12.5. mu.L of bovine serum albumin coated gold nanoclusters (5mM, Au in chloroauric acid)3+Concentration of ion) and 25. mu.L of 1, 2-bis [4- (3-sulfopropoxy) phenyl group]-1, 2-stilbene sodium salt (0.1mg/L) mixed to a fluorescent ratiometric probe. Mixing 100 μ L serum with 100 μ L glucose oxidase (0.5mg/L) solution, adding 100 μ L PBS buffer solution (10mM, pH7), mixing, adding into fluorescence ratio probe, adding water to constant volume to 500 μ L, mixing, reacting at 37 deg.C for more than 30min, scanning fluorescence emission wavelength of 410-850nm with a fluorescence instrument, and recording fluorescence intensity ratio I and I0I and I0The ratio of the fluorescence intensity at an emission wavelength of 680nm to the fluorescence intensity at 490nm at the sample concentration and blank, respectively (F)680/F490) According to the change value (I) of the fluorescence ratio0-I)/I0Linear relationship with glucose concentration C (I)0-I)/I0Blood glucose levels were calculated and compared to hospital traditional method test values at 0.0705C +0.0407, with the results shown in the table below, which is more accurate than traditional thinking.
Example 5
And (3) selective detection of glucose: 12.5. mu.L of bovine serum albumin coated gold nanoclusters (5mM, Au in chloroauric acid)3+Concentration of ion) and 25. mu.L of 1, 2-bis [4- (3-sulfopropoxy) phenyl group]-1, 2-stilbene sodium salt (0.1mg/L) mixed to a fluorescent ratiometric probe. 100 mu L of glucose, sucrose,Mixing fructose, maltose and xylose solution (concentration is 5mM) and 100 μ L glucose oxidase (0.5mg/L) solution, adding 100 μ L PBS buffer solution (10mM, pH7), adding into fluorescence ratio probe, adding water to constant volume to 500 μ L, mixing, reacting at 37 deg.C for 30min, scanning fluorescence emission wavelength of 410-850nm with fluorescence instrument, and recording fluorescence intensity ratio I and I0I and I0The ratio of the fluorescence intensity at an emission wavelength of 680nm to the fluorescence intensity at 490nm at the sample concentration and blank, respectively (F)680/F490) As shown in fig. 5, in the presence of only glucose, the fluorescence intensity ratio varied greatly, and sucrose, fructose, maltose, and xylose interfered less with the detection. The method can realize specific detection of glucose.
In summary, the above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (1)
1. A fluorescence ratiometric probe based on aggregation-induced emission properties for hydrogen peroxide and glucose detection, characterized by: the gold nanoclusters are coated by tetraphenyl ethylene sulfonate and bovine serum albumin; the tetraphenyl ethylene sulfonate is used as a reference fluorescent probe, the gold nanocluster coated by the bovine serum albumin is used as a fluorescent probe, the tetraphenyl ethylene sulfonate and the bovine serum albumin are mixed to construct a fluorescent ratiometric probe, and the color of the mixed solution is red, yellow or blue according to different mixing ratios of the tetraphenyl ethylene sulfonate and the bovine serum albumin; hydrogen peroxide or hydrogen peroxide generated by the reaction of glucose and glucose oxidase can cause the red fluorescence quenching of the gold nanoclusters coated by the bovine serum albumin, the blue-green fluorescence of the tetraphenyl ethylene sulfonate is not influenced, and the detection of the hydrogen peroxide and the glucose is realized according to the change relation between the change value of the fluorescence ratio of the double emission peaks and the concentration of an analyte;
the preparation method of the fluorescence ratio probe based on the aggregation-induced emission property comprises the following specific steps:
step one, preparing a tetraphenyl ethylene sulfonate solution;
step two, preparing gold nanoclusters coated by bovine serum albumin;
step three, mixing the tetraphenyl ethylene sulfonate prepared in the step one with the gold nanoclusters prepared in the step two to obtain a probe; the gold nanoclusters coated by the bovine serum albumin are positively charged, the tetraphenyl vinylsulfonate is negatively charged, the gold nanoclusters coated by the tetraphenyl vinylsulfonate and the bovine serum albumin are subjected to electrostatic adsorption and aggregation induced luminescence, and the fluorescence emission wavelength of the gold nanoclusters is 400-550 nm; the fluorescence emission wavelength of the mixed solution is respectively 400-550nm and 550-850nm, and the ratio type fluorescence probe with dual-wavelength emission is obtained;
the tetraphenylethylene sulfonate in the first step at least contains a tetraphenylethylene molecular structure skeleton and at least contains two sulfonic acid heel groups, and the structural formula is shown as (1):
wherein: r1、R2＝Cl、Br、I、CnH2n+1,n＝0，1，2，3……n；
The method for preparing the gold nanocluster coated by the bovine serum albumin comprises the following steps: adding 5mL of chloroauric acid (5-20mM) into 5mL of bovine serum albumin solution (20-50mM), adjusting the pH to be alkaline, and reacting at 25-60 ℃ for 10-24 hours to obtain bovine serum albumin coated gold nanoclusters (BSA-AuNCs);
step one, the tetraphenyl ethylene sulfonate is 1, 2-bis [4- (3-sulfonic acid propoxy) phenyl]1, 2-diphenylethylene sodium salt (BSPOTPE), with the preparation concentration of 0.1 mg/mL; step two, the concentration of the gold nanoclusters coated by the bovine serum albumin is 5mM (in Au in chloroauric acid)3+Concentration meter of ions); in the third step, the volume ratio of the tetraphenyl ethylene sulfonate solution to the bovine serum albumin coated gold nanoclusters is 1: 0.5;
the semi-quantitative method for detecting hydrogen peroxide or glucose by adopting the fluorescence ratio probe comprises the following steps of respectively adding known solutions to be detected with different concentrations C into a probe solution under the irradiation of 365nm ultraviolet light, recording the color of the probe solution after the color change is stable, then dropwise adding the solution to be detected into the probe solution, indicating that the solution to be detected contains hydrogen peroxide or glucose if the color change of the probe solution is visually observed, and realizing semi-quantitative detection according to the comparison between the color change and the color of a sample to be detected with known concentration;
the ratio of fluorescence intensity I at 680nm and 490nm of the initial probe was recorded0，I0＝F680/F490Respectively adding the known solutions to be detected with different concentrations C into the probe solution, recording the fluorescence intensity ratio I after the color change of the probe solution is stable, and obtaining the change value (I) according to the fluorescence intensity ratio0-I)/I0And the change rule of the concentration C of the solution to be detected to obtain a linear relation equation: (I)0-I)/I0And a is the slope of a linear equation, b is the intercept of the linear equation, and the sample detection with unknown concentration is realized according to the linear equation.
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