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Method used for rapid determination of hydrogen content of water using acetylcholin esterase

A technology of acetylcholinesterase and thioacetylcholine, which is applied in the field of enzymatic analysis in the field of biotechnology, can solve the problems of cumbersome and harsh operations, the inability to apply real-time rapid detection, and the lack of rapid detection kits, etc., and achieve the effect of fast response and simple operation

Active Publication Date: 2014-07-09
BEIJING UNIV OF TECH
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0008] The purpose of the present invention is to seek a fast and sensitive qualitative and semi-quantitative method to solve the problem that the current detection methods of the concentration of hydrogen molecules in sample water need certain instruments and equipment and the operation is cumbersome and harsh, which cannot be applied to real-time rapid detection, and there is currently no rapid Problems with kits for detecting hydrogen concentration in samples

Method used

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  • Method used for rapid determination of hydrogen content of water using acetylcholin esterase

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Embodiment 1

[0021] Accurately weigh 0.029 mg thioacetylcholine and 0.024 mg DTNB respectively, including 0.1 μg of acetylcholinesterase (from electric eel, with a total activity of 827 U / mg), add an appropriate amount of disodium hydrogen phosphate (to make the pH of the final solution system 7) into In a centrifuge tube, after drying, seal and freeze. Use the hydrogen electrode to measure the hydrogen water used to be 3000 μmol / L and 200 μmol / L. Use these two different concentrations of hydrogen water and pure water as solvents, respectively take 200 μL and add them to the centrifuge tube at the same time, mix well, and the visible light absorption value is Detection is carried out at 412nm, the higher the absorption value corresponds to the higher the concentration, see figure 1 .

Embodiment 2

[0023] Accurately weigh 0.029 mg thioacetylcholine and 0.024 mg DTNB, including 0.11 μg of acetylcholinesterase (from electric eel, with a total activity of 827 U / mg), add an appropriate amount of disodium hydrogen phosphate (so that the pH of the final solution is 6.5) and put them into a centrifuge tube After drying, seal and freeze. Use a hydrogen electrode to measure the concentration of the hydrogen water used is 100 μmol / L, use pure water and hydrogen water of this concentration as solvents, respectively take 200 μL and add them to the centrifuge tube at the same time, mix well, and perform detection at the visible light absorption value of 412nm, see figure 2 .

Embodiment 3

[0025] Accurately weigh 0.87 mg thioacetylcholine and 0.24 mg DTNB, including 0.36 μg of acetylcholinesterase (from electric eel, with a total activity of 827 U / mg), add disodium hydrogen phosphate (so that the pH of the final solution is 7.5) and put it into a centrifuge tube , sealed and frozen. Use a hydrogen electrode to measure the concentration of the hydrogen water used is 500 μmol / L, use pure water and hydrogen water of this concentration as solvents, respectively take 200 μL and add them to the centrifuge tube at the same time, mix well, and detect at the visible light absorption value of 412nm, the color The deep one is 500μmol / L concentration.

[0026] Detection steps of the present invention are:

[0027] According to the method of the present invention, accurately weigh 0.03mg of thioacetylcholine and 0.024mg of DTNB, acetylcholinesterase and sodium dihydrogen phosphate respectively, put them into a centrifuge tube, take the same dose of each material in another ...

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Abstract

The invention discloses a method used for rapidly determining that whether hydrogen concentration of water samples is as large as an effective concentration, and belongs to the field of enzymatic analysis of biotechnology. According to the method, acetylcholine esterase, acetylthiocholine, a reaction buffer system reagent, and color developing agent DTNB are mixed uniformly, an obtained mixture is divided into two equal parts; hydrogen water to be tested is added into a first part of the mixture, and deionized water is added into a second part of the mixture, wherein adding amount of the hydrogen water is equal to that of the deionized water, and pH values of obtained solutions are both 7; and after reaction, hydrogen content can be determined by comparing absorbance values at visible light 412nm of the solutions, or based on the shade of the colour of yellow products, wherein the larger the absorbance value of the first part is than that of the second part, or the heavier the colour of a product of the first part is than that of a product of the second part, the higher the hydrogen content is. According to the method, hydrogen content of water can be determined directly via color development reaction of enzymatic determination.

Description

technical field [0001] The invention relates to a method for quickly judging the hydrogen content in water by using acetylcholinesterase, especially capable of realizing the qualitative and rapid identification of hydrogen water and ordinary water, and belongs to the field of enzymatic analysis in the field of biotechnology. Background technique [0002] Hydrogen is a colorless, tasteless, odorless, flammable gas and is the lightest diatomic molecule in nature. In the field of diving medicine, people use hydrogen-oxygen mixture as a breathing medium to shorten the decompression time. For a long time, it has been believed that hydrogen gas is just a metabolic waste gas of intestinal flora and has no biological effect. Therefore, hydrogen has been considered as a physiologically inert gas. In 2007, Japanese scholars published an article in "Natural Science" and found that in vivo models proved that breathing 2% hydrogen can effectively reduce the infarct size caused by cereb...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/78
Inventor 马雪梅王婷婷刘梦昱
Owner BEIJING UNIV OF TECH
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