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Electrochemical method used for measuring activity of alkaline phosphatase

A phosphatase activity, electrochemical technology, applied in the field of medical testing, to achieve the effect of small sample demand, low cost and accurate measurement

Active Publication Date: 2015-03-04
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, a fast, simple and accurate electrochemical method for the determination of alkaline phosphatase activity using pyrophosphate ion as substrate has not been reported

Method used

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  • Electrochemical method used for measuring activity of alkaline phosphatase
  • Electrochemical method used for measuring activity of alkaline phosphatase
  • Electrochemical method used for measuring activity of alkaline phosphatase

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0039] Preparation of the gold electrode modified by embodiment 1, cysteine ​​and copper ions

[0040] 1) Gold electrode pretreatment and activation in sulfuric acid solution

[0041] A gold electrode with a diameter of 1.5mm was used as the working electrode, and the electrode was polished and cleaned before use. The specific process is as follows: the electrode is firstly polished and polished with aluminum oxide powder slurry with a diameter of 0.3 μm and 0.05 μm on the polishing cloth, and then cleaned with secondary deionized water, and ultrasonicated in secondary deionized water and ethanol respectively. Treat for 5 minutes to remove residual Al2O3 powder, and finally clear it with secondary deionized water, and dry it under nitrogen for use. The treated gold electrode was subjected to continuous cyclic voltammetry scanning in 5mL of 0.5M sulfuric acid solution, and the scanning potential window was 0.3V-1.7V.

[0042] figure 2 It is the cyclic voltammetry curve of g...

Embodiment 2

[0047] Example 2, Study on the Selectivity of Cysteine ​​and Copper Ion Modified Gold Electrodes to Different Kinds of Anions

[0048] The gold electrode modified by cysteine ​​and copper ions was placed in 4 mL of 0.05 M phosphate buffer solution for cyclic voltammetry scanning, and after stabilization, 1 mL of 20 μM F - , Cl - 、Br - , CO 3 2- 、HCO 3 - , SO 4 2- 、CH 3 COO - , NO 3 - 、H 2 PO 4 - 、HPO 4 2- , PPi and other different kinds of anions were scanned by cyclic voltammetry.

[0049] Figure 5 The selectivity of the gold electrode modified by the cysteine ​​and copper ions to different kinds of anions.

[0050] Depend on Figure 5 It can be seen that the prepared gold electrode modified by cysteine ​​and copper ions has good selectivity to pyrophosphate ion (PPi).

Embodiment 3

[0051] The standard curve of embodiment 3, gold electrode modified by cysteine ​​and copper ions responding to pyrophosphate ions

[0052] The cysteine ​​and copper ion-modified gold electrodes were placed in 4 mL of 0.05 M phosphate buffer solution for cyclic voltammetry scanning, and after stabilization, 1 mL of 1 μM pyrophosphate ion solution and 1 mL of 5 μM pyrophosphate Ion solution, 1 mL of 10 μM pyrophosphate ion solution, 1 mL of 15 μM pyrophosphate ion solution, 1 mL of 20 μM pyrophosphate ion solution, 1 mL of 25 μM pyrophosphate ion solution, and 1 mL of 30 μM pyrophosphate ion solution, stirred Evenly, the detection solution was obtained, left for 5 min, and subjected to cyclic voltammetry scanning. Figure 6 The standard curve for the pyrophosphate ion response of the cysteine ​​and copper ion modified gold electrode.

[0053] Depend on Figure 6 It can be seen that when the concentration of pyrophosphate ions added is in the range of 5 μM to 25 μM (that is, th...

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Abstract

The invention provides an electrochemical method used for measuring activity of alkaline phosphatase and an electrochemical sensor used in the method. The method comprises the following steps: preparing an electrode modified by cysteine and copper ions; drawing a standard curve between the enzyme activity concentration of alkaline phosphatase and relative current intensity under a pyrophosphate ion substrate with specific concentration; changing the alkaline phosphatase solution with known enzyme activity concentration into a to-be-measured sample containing alkaline phosphatase, performing cyclic voltammetry scanning to obtain the relative current intensity, and calculating to obtain the activity concentration of the alkaline phosphatase in the to-be-measured sample according to the standard curve. According to the method disclosed by the invention, a reversible competitive coordination principle of the cysteine, pyrophosphate ions and copper ions is utilized, the electrochemical sensor used for measuring the activity of alkaline phosphatase is prepared, and the activity of alkaline phosphatase can be rapidly, simply and accurately measured by utilizing the electrochemical sensor.

Description

technical field [0001] The invention belongs to the technical field of medical testing, and in particular relates to an electrochemical method for measuring the activity of alkaline phosphatase. Background technique [0002] Alkaline phosphatase (ALP) is a very common biological enzyme in organisms, widely present in various parts of organisms (such as: small intestine, liver, bones, kidneys, placenta, etc.). Normal adult serum alkaline phosphatase content is 40 ~ 150IU / L. Its main function is to hydrolyze and dephosphorylate the corresponding substrates to maintain normal metabolism in the organism. Studies have shown that too high or too low alkaline phosphatase can cause a series of diseases, such as: primary liver cancer, thymus cancer, breast cancer, prostatitis, bone cell cancer and diabetes and so on. Therefore, being able to quickly and simply measure alkaline phosphatase activity has important physiological and pathological significance in clinical medicine. ...

Claims

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Application Information

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IPC IPC(8): G01N27/48G01N27/30
Inventor 毛兰群姜琴
Owner INST OF CHEM CHINESE ACAD OF SCI
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