Fluorescent sensor for detecting inorganic pyophosphatase and its preparation method

A technology of inorganic pyrophosphatase and fluorescent sensor, applied in chemical instruments and methods, fluorescence/phosphorescence, instruments, etc., can solve the problems of low fluorescence intensity, time-consuming process, expensive instruments, etc., and achieve the effect of efficient detection methods

Active Publication Date: 2018-05-18
FUZHOU UNIV
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Problems solved by technology

However, these methods have disadvantages such as expensive instruments, time-consuming process, poor selectivity, and potential harm to human body.
[0003] CN104237193A discloses a fluorescent sensor for detecting pyrophosphatase and its preparation method. When no pyrophosphatase is added and only pyrophosphate exists, pyrophosphate and divalent copper form a complex, and sodium ascorbate cannot reduce the complexed divalent copper. The valence copper is monovalent copper as a catalyst, and azidocoumarin and propynyl alcohol cannot undergo 1,3-dipolar cycloaddition reaction, so the fluorescence intensity of the system is very low; but in the case of adding pyrophosphatase at the same time , pyrophosphatase can hydrolyze pyrophosphate to inhibit the formation of divalent copper complexes, the click chemical reaction can continue to occur, the fluorescence intensity of the system is greatly enhanced and there is a linear relationship between the fluorescence intensity and the amount of pyrophosphatase added
The sensor has been successfully applied to the detection of sodium fluoride, a pyrophosphatase inhibitor, but this method requires the preparation of azidocoumarin, which requires the addition of high-energy azide, which has certain risks.

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  • Fluorescent sensor for detecting inorganic pyophosphatase and its preparation method
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  • Fluorescent sensor for detecting inorganic pyophosphatase and its preparation method

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preparation example Construction

[0034] A method for preparing a fluorescent sensor for detecting inorganic pyrophosphatase, comprising the following steps:

[0035] 1) Prepare inorganic pyrophosphatase solutions with different concentration units;

[0036] 2) Disperse the two-dimensional metal-organic framework Cu-BDC-MOF nanosheets with simulated enzyme activity in ultrapure water and sonicate for 30 min, centrifuge to take the supernatant for later use; the Cu-BDC-MOF nanosheets in the supernatant The concentration is 200 mg / mL;

[0037] 3) Dissolve sodium pyrophosphate in tris(hydroxymethyl)aminomethane-hydrochloride buffer solution to prepare sodium pyrophosphate solution, store at 4°C for later use;

[0038] 4) Dilute 60 times with 30% hydrogen peroxide solution and store at 4°C for later use;

[0039] 5) Dissolve terephthalic acid in tris(hydroxymethyl)aminomethane-hydrochloric acid buffer to prepare a terephthalic acid solution, and store it in the dark at 4°C for later use;

[0040] 6) Take 20 μL ...

Embodiment 1

[0050] A method for preparing a fluorescent sensor for detecting inorganic pyrophosphatase, the specific steps are:

[0051] 1. Prepare inorganic pyrophosphatase solutions with different concentration units;

[0052] 2. First synthesize Cu-BDC-MOF nanosheet material, the steps are as follows:

[0053] 1) Dissolve 3.0 mg of terephthalic acid in 3.0 mL of a mixed solvent of N,N-dimethylformamide and acetonitrile with a volume ratio of 2:1;

[0054] 2) Dissolve 3.0 mg of copper nitrate trihydrate in 3.0 mL of a mixed solvent of N,N-dimethylformamide and acetonitrile with a volume ratio of 1:2;

[0055] 3) Prepare 2.0 mL of a mixed solvent of N,N-dimethylformamide and acetonitrile with a volume ratio of 1:1 as the linking solution;

[0056] 4) First add terephthalic acid solution into a test tube with a diameter of 2.0 cm and a capacity of 30 mL, then add 2.0 mL of connecting solution, and finally add copper nitrate solution; Heated and reacted for 36 hours; the blue product wa...

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Abstract

The invention belongs to the field of applying a metal organic framework to analysis and detection of a disease marker, and particularly relates to a fluorescent sensor for detecting inorganic pyophosphatase and its preparation method. Through reacting different concentrations of inorganic pyophosphatase solution with pyrophosphate, Cu-BDC-MOF nanosheet solution, hydrogen peroxide solution and terephthalic acid solution are added and incubated at 37 DEG C for 30 minutes; fluorescent signals of inorganic pyophosphatase with different concentrations is detected, a working curve is structured, and the fluorescent sensor is prepared. The fluorescent signal of the inorganic pyophosphatase in a sample is detected by the fluorescent sensor, and then corresponding to the working curve, thus the quantitative analysis of the inorganic pyophosphatase in the sample is realized. The detection method is high in selectivity, fast, sensitive and high-efficient.

Description

technical field [0001] The invention belongs to the field of analysis and detection of metal organic framework materials used for disease markers, and in particular relates to a fluorescent sensor for detecting inorganic pyrophosphatase and a preparation method thereof. Background technique [0002] Pyrophosphatase (EC 3.6.1.1), or inorganic pyrophosphatase (PPase), is a hydrolase that specifically converts pyrophosphate (P2O74-, PPi) into inorganic phosphate. The degradation of pyrophosphate (PPi) by inorganic pyrophosphatase (PPase) is a massively exothermic reaction, which provides the thermodynamic drive for biosynthetic reactions (J. Biomol. Screening 2013, 18, 490−497.) . Early studies have shown that inorganic pyrophosphatase (PPase) is directly related to phosphorus metabolism, carbohydrate metabolism and evolution (Biochim. Biophys. Acta 2006, 1764, 1299−1306.). Thus, PPase is essential for energy metabolism processes that occur in living organisms, and depletion ...

Claims

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Application Information

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IPC IPC(8): G01N21/64C09K11/06
CPCC09K11/06C09K2211/188G01N21/6428
Inventor 邱彬胡水生郭隆华林振宇陈国南
Owner FUZHOU UNIV
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