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Fluorescent probe for quantitatively detecting riboflavin on basis of fluorescence resonance energy transfer ratio, and preparation method and application for fluorescent probe

A fluorescence resonance energy, quantitative detection technology, applied in the field of fluorescent probes, can solve the problems of weak anti-interference ability, cumbersome steps, time-consuming and laborious, etc., and achieve high accuracy and sensitivity, simple preparation method, and strong anti-interference ability. Effect

Active Publication Date: 2020-05-08
SHANXI UNIV
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  • Abstract
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Problems solved by technology

[0003] The methods for detecting riboflavin that have been reported so far include: electrochemical method, spectrophotometry, immunoassay, capillary electrophoresis and high performance liquid chromatography, etc. Although these methods have their own advantages, they also have certain defects. , such as cumbersome steps, time-consuming and laborious, complex and expensive instruments, long detection cycle, high cost, weak anti-interference ability and low accuracy, etc., which limit the practical application of the above detection methods

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  • Fluorescent probe for quantitatively detecting riboflavin on basis of fluorescence resonance energy transfer ratio, and preparation method and application for fluorescent probe
  • Fluorescent probe for quantitatively detecting riboflavin on basis of fluorescence resonance energy transfer ratio, and preparation method and application for fluorescent probe
  • Fluorescent probe for quantitatively detecting riboflavin on basis of fluorescence resonance energy transfer ratio, and preparation method and application for fluorescent probe

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Preparation and characterization of NPCl-CQDs

[0030] Step 1: Weigh 0.4 g of glucose, add 6 mL of ethylenediamine, 2 mL of concentrated phosphoric acid and 2 mL of concentrated hydrochloric acid to it to obtain a brown viscous substance.

[0031] Step 2: After the beaker was cooled to room temperature, 30 mL of ultrapure water was added to it, stirred to fully dissolve, centrifuged at 4000 rpm for 10 minutes, and the supernatant was filtered with a filter membrane to obtain a clear NPCl-CQDs solution.

[0032] Step 3, freeze-drying to obtain solid powder of NPCl-CQDs.

[0033] Step 4: Weigh 0.1 g of NPCl-CQDs solid powder into a beaker, add 10 mL of ultrapure water to it, and ultrasonically dissolve it fully to obtain a stock solution of NPCl-CQDs with a concentration of 10 mg / mL.

[0034] Characterization see figure 1 and figure 2 . figure 1 In the UV absorption spectrum of NPCl-CQDs, there are two obvious absorption peaks located at 279 nm and 317 ...

Embodiment 2

[0035] Embodiment 2: Anti-interference experiment of riboflavin detection

[0036] Step 1: Accurately weigh a certain mass of metal ions (Fe 3+ , K + , Fe 2+ , Mg 2+ , Cu 2+ , Al 3+ , Ag + , Ca 2+ , Cr 3+ , Na + , Mn 2+ , Zn 2+ , Hg 2+ , Cd 2+ , Ba 2+ ,Co 2+ ) compound, add 10 mL of ultrapure water, and ultrasonically dissolve to prepare a metal ion stock solution with a concentration of 0.1 mol / L.

[0037] Step 2: Accurately weigh a certain mass of amino acids (valine, phenylalanine, threonine, arginine, alanine, leucine, isoleucine, tryptophan, glycine, serine, semi- cystine, tyrosine, histidine, lysine, glutamic acid, aspartic acid, glutamine, methionine, proline, homocysteine, glutathione), Add 5 mL of ultrapure water, ultrasonically dissolve, and prepare an amino acid stock solution with a concentration of 0.01 mol / L.

[0038] Step 3, accurately weigh a certain mass of anions (Br - , I - , F - , CO 3 2- , S 2 o 3 2- , HCO 3 2- , SO 3 2- , NO ...

Embodiment 3

[0045] Embodiment 3: the linear equation of riboflavin titration NPCl-CQDs

[0046] Step 1, measure the fluorescence intensity of NPCl-CQDs (0.57 mg / mL), record the fluorescence intensity of NPCl-CQDs at 530 nm and 453 nm, respectively, denoted as I 530 nm and I 453 nm , and calculate the ratio I 530 nm / I 453 nm .

[0047] Step 2: Add riboflavin stock solution drop by drop, and record the fluorescence intensity of the mixed solution of NPCl-CQDs and riboflavin at 530 nm and 453 nm, respectively, as I 530 nm and I 453 nm , and calculate its ratio I 530 nm / I 453 nm . Changes in fluorescence intensity see Figure 7 .

[0048] Step 3, using Origin software, fitting the fluorescence intensity ratio of NPCl-CQDs and riboflavin mixed solution at 530 nm and 453 nm ( I 530 nm / I 453 nm ) and riboflavin concentration, a linear equation is obtained, and the results are shown in Figure 8 and Figure 9 .

[0049] Figure 7 It shows that with the addition of ribo...

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Abstract

The invention belongs to the technical field of fluorescent probes, and provides a fluorescent probe for quantitatively detecting riboflavin on the basis of a fluorescence resonance energy transfer ratio, and a preparation method and an application for the fluorescent probe. The preparation method comprises the following steps: with glucose as a carbon source, ethylenediamine as a nitrogen source,concentrated phosphoric acid as a phosphorus source and concentrated hydrochloric acid as a chlorine source, preparing nitrogen-phosphorus-chlorine co-doped carbon quantum dots (NPCl-CQDs) through anacid-base neutralization reaction exothermic carbonization method, dissolving NPCl-CQDs powder into ultrapure water, carrying out centrifuging to remove insoluble substances, carrying out filtering to remove impurities, and carrying out freeze-drying so as to obtain the fluorescent probe, namely NPCl-CQDs solid powder. The linear relationship between a riboflavin concentration and an NPCl-CQDs fluorescence intensity is determined by using a ratio fluorescence detection method. The content of riboflavin in an actual sample is detected through a standard recovery experiment, and a standard recovery rate is calculated. The method provided by the invention is simple and convenient to operate, low in background interference, high in sensitivity, free of expensive instruments and equipment, lowin detection cost, capable of rapidly, efficiently and quantitatively detecting the content of the riboflavin in an actual sample in a ratio and good in reproducibility.

Description

technical field [0001] The invention belongs to the technical field of fluorescent probes, and in particular relates to a fluorescent probe for quantitatively detecting riboflavin based on a fluorescence resonance energy transfer ratio and a preparation method and application thereof. Background technique [0002] Riboflavin, the B vitamin 2 (Vitamin B 2 , VB 2 ), is one of the 13 essential vitamins for the human body, it can participate in biological oxidation and energy metabolism in organisms, promote growth and development, maintain the integrity of skin and cell membranes, and protect the skin, hair follicles, mucous membranes and sebaceous glands; it can participate in the growth of cells Metabolism is an essential nutrient for the metabolism and repair of body tissues; it can participate in vitamin B 6 and niacin (VB 3 ) metabolism, is a model of coordination of B vitamins. When there is a lack of riboflavin in the organism, it will affect the biological oxidatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/65C09K11/72C01B32/15B82Y20/00B82Y30/00B82Y40/00G01N21/64
CPCB82Y20/00B82Y30/00B82Y40/00C09K11/65C09K11/72C01B32/15G01N21/6428G01N2021/6439
Inventor 弓晓娟王子涵董文娟刘洋董川
Owner SHANXI UNIV
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