Detection method for heme based on boron-doped grapheme quantum dots fluorescence quenching effect

Abstract

The invention discloses a detection method for heme based on the fluorescence quenching effect of boron-doped grapheme quantum dots (B-GQDs). The detection method comprises the following steps: 1) preparing a B-GQDs solution; 2) preparing a heme solution; 3) preparing a standard solution; 4) creating a working curve for detecting the concentration of the heme with the B-GQDs; and 5) detecting the concentration of the heme in a sample to be detected. According to the detection method, based on the fluorescence quenching effect of the B-GQDs, the B-GQDs are firstly applied to detection for the heme, and the heme can be quickly and efficiently detected by using a fluorescence analyzing method. The detection method has the advantages of high response speed, high sensitivity, simplicity in operation, low cost, strong optics antijamming capability and the like; the content of the heme can be detected through the change of a fluorescence signal, so as to realize quick and sensitive qualitative detection and quantitative determination for the biochemical index of the heme.

Description

technical field

[0001] The invention belongs to the technical field of biological fluorescence detection, and relates to a heme detection method, in particular to a heme detection method based on the fluorescence quenching effect of boron-doped graphene quantum dots. Background technique

[0002] Heme is a natural iron porphyrin, which is the active center of hemoglobin and myoglobin in organisms. It mainly exists in the blood and muscles of organisms. It has important physiological functions and high practical value. It is used in medicine, food, It is widely used in chemical industry and other industries. Heme can not only be used as a coloring agent to replace nitrite and synthetic pigments, but also be used to prepare anticancer drugs and anti-anemia drugs. It is currently recognized as the biological iron agent with the highest absorption rate and the best effect.

[0003] In blood testing, in addition to testing blood sugar and cholesterol, hemoglobin is also an impor...

Examples

Embodiment 1B

[0032] Embodiment 1B-GQDs preparation and solution preparation

[0033] 1. Preparation of boron-doped graphene quantum dots

[0034] Boron-doped graphene quantum dots (B-GQDs) were synthesized by potentiostatic chronoamperometry via an electrochemical workstation (CHI660B). The specific method is: prepare 0.1mol / L borax aqueous solution, and take 20mL as electrolyte. A high-purity graphite rod was used as the working electrode, a platinum wire electrode was used as the counter electrode, and a calomel electrode was used as the reference electrode, and the working potential was 3V. After reacting for 2 hours, the solution was filtered through a 0.22 μm filter membrane to remove graphene sheets. Dialyze through a 3500Da dialysis bag to remove Na from the solution + and B 4 o 7 2- , to get B-GQDs.

[0035] 2. Prepare boron-doped graphene quantum dot solution: disperse the above-mentioned synthetic B-GQDs in phosphate buffer (concentration is 0.1mol / L, pH value is 7.4) to o...

Embodiment 2B

[0037] Embodiment 2B-GQDs fluorescence performance evaluation

[0038] The fluorescent performance of B-GQDs solution was detected by fluorescence spectrometer; the excitation wavelength was 380nm, and the emission wavelength was 530nm.

[0039] The quantum dot solution prepared in Example 1 was diluted with phosphate buffer (0.1 mol / L, pH 7.4) until the B-GQDs concentration was 15 μg / mL, and the fluorescence spectrum of the solution was detected;

[0040]Take a certain amount of the quantum dot solution prepared in Example 1 and mix it with the heme solution, add phosphate buffer (concentration is 0.1mol / L, pH value is 7.4) to dilute to obtain a mixed solution; wherein the concentration of B-GQDs is 15 μg / mL , the hemoglobin concentration was 1000 μmol / L. The fluorescence spectrum of the mixed solution was detected by a fluorescence spectrometer.

[0041] like image 3 As shown, curve a in the figure is the fluorescence signal of the B-GQDs blank sample without heme, and b...

Embodiment 3

[0042] Embodiment 3 establishes the working curve of B-GQDs detection hemoglobin concentration

[0043] Take a certain amount of quantum dot solution and heme solution prepared in Example 1, mix and add phosphate buffer (concentration is 0.1mol / L, pH value is 7.4) to dilute, and prepare a group of samples including blank standard sample. Standard solutions with different hemoglobin concentrations; the concentration of B-GQDs is 15 μg / mL, the hemoglobin concentration of the blank standard sample is 0 μmol / L, and the hemoglobin concentration of other standard solutions is 10-1000 μmol / L; a series of different concentrations are detected by fluorescence spectrometer Fluorescence signals of standard solutions of heme and B-GQDs (15 μg / mL), such as Figure 4 As shown, the a-n hemoglobin concentrations are 0μmol / L, 10μmol / L, 50μmol / L, 100μmol / L, 150μmol / L, 200μmol / L, 300μmol / L, 400μmol / L, 600μmol / L, 800μmol / L, 1000μmol / L, 1200μmol / L, 1400μmol / L, 1500μmol / L.

[0044] The response s...

technical field

[0001] The invention belongs to the technical field of biological fluorescence detection, and relates to a heme detection method, in particular to a heme detection method based on the fluorescence quenching effect of boron-doped graphene quantum dots. Background technique

[0002] Heme is a natural iron porphyrin, which is the active center of hemoglobin and myoglobin in organisms. It mainly exists in the blood and muscles of organisms. It has important physiological functions and high practical value. It is used in medicine, food, It is widely used in chemical industry and other industries. Heme can not only be used as a coloring agent to replace nitrite and synthetic pigments, but also be used to prepare anticancer drugs and anti-anemia drugs. It is currently recognized as the biological iron agent with the highest absorption rate and the best effect.

[0003] In blood testing, in addition to testing blood sugar and cholesterol, hemoglobin is also an impor...

Examples

Embodiment 1B

[0032] Embodiment 1B-GQDs preparation and solution preparation

[0033] 1. Preparation of boron-doped graphene quantum dots

[0034] Boron-doped graphene quantum dots (B-GQDs) were synthesized by potentiostatic chronoamperometry via an electrochemical workstation (CHI660B). The specific method is: prepare 0.1mol / L borax aqueous solution, and take 20mL as electrolyte. A high-purity graphite rod was used as the working electrode, a platinum wire electrode was used as the counter electrode, and a calomel electrode was used as the reference electrode, and the working potential was 3V. After reacting for 2 hours, the solution was filtered through a 0.22 μm filter membrane to remove graphene sheets. Dialyze through a 3500Da dialysis bag to remove Na from the solution + and B 4 o 7 2- , to get B-GQDs.

[0035] 2. Prepare boron-doped graphene quantum dot solution: disperse the above-mentioned synthetic B-GQDs in phosphate buffer (concentration is 0.1mol / L, pH value is 7.4) to o...

Embodiment 2B

[0037] Embodiment 2B-GQDs fluorescence performance evaluation

[0038] The fluorescent performance of B-GQDs solution was detected by fluorescence spectrometer; the excitation wavelength was 380nm, and the emission wavelength was 530nm.

[0039] The quantum dot solution prepared in Example 1 was diluted with phosphate buffer (0.1 mol / L, pH 7.4) until the B-GQDs concentration was 15 μg / mL, and the fluorescence spectrum of the solution was detected;

[0040]Take a certain amount of the quantum dot solution prepared in Example 1 and mix it with the heme solution, add phosphate buffer (concentration is 0.1mol / L, pH value is 7.4) to dilute to obtain a mixed solution; wherein the concentration of B-GQDs is 15 μg / mL , the hemoglobin concentration was 1000 μmol / L. The fluorescence spectrum of the mixed solution was detected by a fluorescence spectrometer.

[0041] like image 3 As shown, curve a in the figure is the fluorescence signal of the B-GQDs blank sample without heme, and b...

Embodiment 3

[0042] Embodiment 3 establishes the working curve of B-GQDs detection hemoglobin concentration

[0043] Take a certain amount of quantum dot solution and heme solution prepared in Example 1, mix and add phosphate buffer (concentration is 0.1mol / L, pH value is 7.4) to dilute, and prepare a group of samples including blank standard sample. Standard solutions with different hemoglobin concentrations; the concentration of B-GQDs is 15 μg / mL, the hemoglobin concentration of the blank standard sample is 0 μmol / L, and the hemoglobin concentration of other standard solutions is 10-1000 μmol / L; a series of different concentrations are detected by fluorescence spectrometer Fluorescence signals of standard solutions of heme and B-GQDs (15 μg / mL), such as Figure 4 As shown, the a-n hemoglobin concentrations are 0μmol / L, 10μmol / L, 50μmol / L, 100μmol / L, 150μmol / L, 200μmol / L, 300μmol / L, 400μmol / L, 600μmol / L, 800μmol / L, 1000μmol / L, 1200μmol / L, 1400μmol / L, 1500μmol / L.

[0044] The response s...

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