A preparation method of ratiometric fluorescent probe for quantitative detection of ppi content in urine

A ratiometric fluorescent probe and fluorescent probe technology, applied in the field of detection of PPi in urine, can solve problems such as detection performance limitations

Active Publication Date: 2021-09-14
NANJING MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of these detection methods rely entirely on a single fluorescence intensity signal output, and their detection performance has certain limitations
To the best of our knowledge, there are currently no ratiometric fluorescent probes based on CDs for the detection of PPi

Method used

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  • A preparation method of ratiometric fluorescent probe for quantitative detection of ppi content in urine
  • A preparation method of ratiometric fluorescent probe for quantitative detection of ppi content in urine
  • A preparation method of ratiometric fluorescent probe for quantitative detection of ppi content in urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 The synthesis of blue fluorescent B-CDs, the steps are as follows:

[0031] Weigh 300mg of sodium citrate and 30mg of glutathione, add 10ml of water, ultrasonicate to fully dissolve it, transfer the solution to a 30ml reactor for sealing, and react at 200°C under high temperature and high pressure for 4h, the obtained After cooling, the solution was dialyzed with a 500Da dialysis bag for 12 h to obtain blue fluorescent B-CDs. Its transmission electron microscope image is as follows figure 1 In the middle A, the UV absorption and fluorescence emission diagrams are shown in figure 1 shown in B.

Embodiment 2

[0032] Embodiment 2 Synthesis of Y-CDs with yellow fluorescence, the steps are as follows:

[0033] Weigh 100 mg of 1,2,4-triaminobenzene, add 10 ml of formamide solution, stir to make 1,2,4-triaminobenzene evenly dispersed in the formamide solution, transfer the solution to a 30 ml reaction kettle, The reaction was carried out at 120 °C for 12 h, the solution obtained was centrifuged to remove unreacted substances, the supernatant was collected and purified by dialysis with a 500 Da dialysis bag for 12 h, and the obtained product was filtered with a 0.22 μm microporous membrane to obtain yellow fluorescent Y- CDs solution. Its transmission electron microscope image is as follows figure 2 In the middle A, the UV absorption and fluorescence emission diagrams are shown in figure 2 shown in B.

Embodiment 3

[0034] Example 3 Investigating the effect of B-CDs on Fe 3+ To respond, the steps are as follows:

[0035] Disperse 10 μL of B-CDs solution in 180 μL of pH=4 Tris-HCl buffer, and then add 10 μL of Fe with different concentrations 3+ The final concentration of B-CDs was 50 μM, 100 μM and 200 μM. After incubation at room temperature for 10 min, the fluorescence intensity of the solution was detected to investigate the effect of B-CDs on Fe 3+ response, the experimental results are as image 3 shown.

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Abstract

The invention discloses a preparation method of a ratio fluorescent probe for quantitative detection of PPi content in urine. (1) Using sodium citrate as a carbon source and glutathione as a passivating agent to synthesize B‑CDs emitting blue fluorescence; (2) Using 1,2,4‑triaminobenzene as a carbon source and formamide as Passivator, synthesize Y-CDs that emit yellow fluorescence, and mix it with synthetic B-CDs that emit blue fluorescence to form a ratiometric fluorescent probe that can simultaneously emit blue fluorescence and yellow fluorescence under the same excitation wavelength needle; (3) adding Fe to the probe 3+ , the blue fluorescence of the probe is quenched, and the yellow fluorescence remains unchanged; (4) PPi is added to combine with the Fe in the probe 3+ , the blue fluorescence recovers, and the yellow fluorescence remains unchanged. The concentration of PPi is quantitatively detected by detecting the ratio of the blue fluorescence to the yellow fluorescence, which is sensitive, convenient and selective.

Description

technical field [0001] The invention belongs to the fields of nanomaterials, fluorescence sensing technology and biological analysis and detection, and in particular relates to a method for detecting PPi in urine based on a ratio fluorescence sensing platform based on carbon dots. Background technique [0002] Kidney stones are a high incidence of disease in today's society. The main pathogenic mechanism is the crystallization reaction of insoluble components in the urethra. The main chemical components that form this pathological crystal are calcium oxalate, calcium phosphate and uric acid. Pyrophosphate (PPi, P 2 O 4 7- ) is released in the body during the hydrolysis of adenosine triphosphate to adenosine monophosphate, and is an inhibitor of some crystallization reactions in the body such as the formation of calcium phosphate. Studies have shown that PPi can act as an inhibitor of kidney stone formation, and the content of PPi in patients with kidney stones is related ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C09K11/65G01N21/64B82Y20/00B82Y40/00
Inventor 岑瑶胡琴徐晓曼许贯虹魏芳弟
Owner NANJING MEDICAL UNIV
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