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Application of BUIPTP (binuclear uranyl-isophthalaldehyde-tetrapyrrole) to ATP (Adenosine Triphosphate) analysis

A technology of uranyl complex and dinuclear uranyl, which is applied in the application field of dinuclear uranyl complex in ATP analysis, can solve the problems of high requirements for instruments and equipment, low precision, complicated operation and the like, and achieves good detection effect and low detection. Out-of-limit, high-precision effects

Inactive Publication Date: 2018-11-09
NANHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The existing ATP detection methods are generally cumbersome to operate, time-consuming, complicated preparation process, high requirements for equipment, low precision and other shortcomings

Method used

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  • Application of BUIPTP (binuclear uranyl-isophthalaldehyde-tetrapyrrole) to ATP (Adenosine Triphosphate) analysis
  • Application of BUIPTP (binuclear uranyl-isophthalaldehyde-tetrapyrrole) to ATP (Adenosine Triphosphate) analysis
  • Application of BUIPTP (binuclear uranyl-isophthalaldehyde-tetrapyrrole) to ATP (Adenosine Triphosphate) analysis

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Experimental program
Comparison scheme
Effect test

preparation example Construction

[0036] The preparation process of BUIPTP among the present invention is as follows:

[0037] (1) Add 0.12mol·L to a 250mL round bottom flask -1 100mL of hydrochloric acid, 60mmol of pyrrole and 5mmol of the corresponding aromatic aldehyde were added, and the reaction was carried out at room temperature and protected from light under magnetic stirring. The reaction system was monitored in real time by TLC. Control the appropriate reaction time, when TLC monitors that the reaction substrate aromatic aldehyde is almost completely consumed, the reaction is ended, the reaction solution is neutralized to pH=8-9 with ammonia water, filtered, and continuously washed with petroleum ether until the filtrate is colorless. Send a little of the same body obtained by suction filtration to HPLC to monitor the content of each component. The rest of the solid can be recrystallized with ethyl acetate-petroleum ether to obtain pure dipyrromethane, and then dried under reduced pressure to obtain ...

Embodiment 1

[0039] Embodiment 1: Determination of optimal conditions for detecting ATP

[0040] (1) Add 3mL of 500nmol L to a 10mL colorimetric tube -1 ATP solution.

[0041] (2) Preparation of BUIPTP detection base solution:

[0042] Accurately weigh 0.7400g of BUIPTP solid with an analytical balance, add twice-distilled water to dissolve, transfer to a 100mL volumetric flask, add water to make up to the mark, shake well to obtain 10μmol L -1 The BUIPTP detection base solution.

[0043] (3) Preparation of standard solution: 5 mL of 10 μmol L -1 The BUIPTP detection base solution was added dropwise to the solution in the above step (1).

[0044] 1.1 Determination of the maximum excitation wavelength

[0045] Measure the secondary scattering spectrum intensity (I SOS ), the comparison shows that the maximum excitation wavelength is 283nm, and the maximum emission wavelength is 566nm.

[0046] 1.2 Determination of optimal pH

[0047] Prepare a series of buffer solutions with a pH of...

Embodiment 2

[0050] Embodiment 2: secondary scattering experiment

[0051] (1) Add 3mL of ATP solution and 2mL of Tris-HCl buffer (pH=7) into a 10mL colorimetric tube to obtain ATP concentrations (c) of 0, 50, 100, 200, 300, 400 and 500nmol·L -1 of the seven solutions to be tested.

[0052] (2) Preparation of BUIPTP detection base solution:

[0053] Accurately weigh 0.7400g of BUIPTP solid with an analytical balance, add twice-distilled water to dissolve, transfer to a 100mL volumetric flask, add water to make up to the mark, shake well to obtain 10μmol L -1 The BUIPTP detection base solution.

[0054] (3) Preparation of standard solution: 5 mL of 10 μmol L -1 The BUIPTP detection base solution was added dropwise to each solution to be tested to obtain the corresponding seven standard solutions. The total dropping time was 10 minutes and incubated for 30 minutes.

[0055] Among them, we have learned through experiments that the secondary scattering signal is stronger when BUIPTP is add...

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Abstract

The invention discloses application of a BUIPTP (binuclear uranyl-isophthalaldehyde-tetrapyrrole) to ATP (Adenosine Triphosphate) analysis and belongs to the technical field of biochemical analysis detection method. The application comprises the following steps: determining optimal conditions for detecting ATP; preparing a BUIPTP detection base solution; drawing a standard curve of determined ATP;analyzing the content of the ATP in a sample: weighing a sample to be detected, which contains the ATP; adding water for dissolving or diluting; adding a Tris-HCl buffering solution; while stirring,dropwise adding the BUIPTP detection base solution into the solution; determining second-grade scattering spectral intensity at a part with optimal wavelength. By adopting a detection method providedby the invention, the deviation between the detected ATP concentration and objective concentration is less than 2.5 percent; the detection method provided by the invention has very high accuracy; whenthe concentration of the ATP is lower than 0.75nmol.L<-1>, the ATP still can be detected; the detection method has a good detection effect in a range of 2.5 to 500nmol.L<-1>.

Description

technical field [0001] The invention relates to the technical field of biochemical analysis and detection methods, in particular to the application of a dinuclear uranyl complex in ATP analysis. Background technique [0002] Adenosine triphosphate (ATP) is a high-energy compound, which is the direct source of energy required for all life activities of tissue cells in organisms. It can store and transfer chemical energy and participate in the metabolism of proteins, fats, sugars and nucleic acids in the body. ATP plays an important role in various physiological and pathological processes of cells, and can be used as an important marker of cell activity. ATP is also often used as an indicator to detect microbial contamination. [0003] The existing ATP detection methods generally have the disadvantages of cumbersome operation, long time-consuming, complicated preparation process, high requirements for instruments and equipment, and low precision. Therefore, it is of great si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/64
CPCG01N21/31G01N21/64
Inventor 肖锡林许丽蒋敏廖力夫王娇彭鹏程
Owner NANHUA UNIV
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