Method for measuring beta-hydroxybutyric acid by using enzyme colorimetric reaction, matched kit and application thereof

A colorimetric reaction, hydroxybutyric acid technology, applied in the field of medical testing and determination, can solve problems such as exogenous interference, and achieve the effects of short reaction time, simple operation, and prevention of complications

Active Publication Date: 2010-06-09
NINGBO RUI BIO TECH
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  • Abstract
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Problems solved by technology

[0004] However, this method is susceptible to interference from exogenous

Method used

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  • Method for measuring beta-hydroxybutyric acid by using enzyme colorimetric reaction, matched kit and application thereof
  • Method for measuring beta-hydroxybutyric acid by using enzyme colorimetric reaction, matched kit and application thereof
  • Method for measuring beta-hydroxybutyric acid by using enzyme colorimetric reaction, matched kit and application thereof

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Embodiment Construction

[0026] The following are specific examples of the present invention to further describe the technical solutions of the present invention, but the present invention is not limited to these examples.

[0027] The enzyme colorimetric reaction of the present invention measures β-hydroxybutyric acid is a one-step enzyme reaction coupled on the basis of enzyme kinetics reaction to determine β-hydroxybutyric acid. In the reaction process, first β-hydroxybutyric acid is Under the action of butyrate dehydrogenase, acetoacetate, reduced coenzyme I and hydrogen ions are generated, and the generated reduced coenzyme I and iodonitrotetrazolium violet react under the catalysis of diaphorase to form the red substance formazane ) has the highest absorbance at 505nm, and its reaction equation is as follows:

[0028]

[0029]

[0030]

[0031] 1. Reagent composition:

[0032] Reagent I includes 100mmol / L Tris-HCL buffer, 20mmol / L oxalic acid with a pH value of 8.5, 2.5mmol / L coenzyme ...

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Abstract

The invention provides a method for measuring beta-hydroxybutyric acid by using enzyme colorimetric reaction, a matched kit and application thereof. Under the system of Tris-HCL buffer solution, serum beta-hydroxybutyric acid and coenzyme I are dehydrogenized under the catalysis of beta-hydroxybutyric acid dehydrogenase to generate acetoacetic acid and reduced coenzyme I, the generated reduced coenzyme I and iodonitrotetrazole chloride are reacted under the catalysis of diaphorase to generate a red substance formazane of which the highest absorbance is 505 nanometers, and then the content of the beta-hydroxybutyric acid in a biological sample is quantified by measuring the change of the absorbance of the red product at the wavelength of 505 nanometers. The method can linearly measure the concentration range (0 to 4.5 mmol / L) of the beta-hydroxybutyric acid in the biological sample, the measurement is used for judging the human ketosis for acid poisoning diagnosis, the reagent used by the method is liquid dual-reagent, and the quantity of the sample required for measuring the beta-hydroxybutyric acid is little; moreover, the reaction time during measuring is short, the operation is simple, and the method is suitable for mass detection.

Description

technical field [0001] The invention belongs to the technical field of medical examination and measurement, and in particular relates to a method for measuring β-hydroxybutyric acid by using an enzyme colorimetric reaction, a matched kit and its application. Background technique [0002] The current detection method for the determination of serum β-hydroxybutyric acid is the enzyme kinetic method, in the coenzyme I (NAD + ) in the presence of β-hydroxybutyric acid under the action of β-hydroxybutyrate dehydrogenase to generate acetoacetate, reduced coenzyme I (NADH) and hydrogen ions, and the reduced coenzyme I (NADH) produced by the reaction is at 340nm The change of absorbance can indirectly reflect the concentration of β-hydroxybutyric acid in the patient's serum. Its reaction process is as follows: [0003] [0004] However, this method is susceptible to exogenous interference. Contents of the invention [0005] The technical problem to be solved by the present i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31C12Q1/32C12Q1/26
Inventor 张闻
Owner NINGBO RUI BIO TECH
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