Determining method and determining kit for beta-hydroxybutyrate

A technology of oxybutyric acid and a kit, which is applied in the field of medical testing and determination, can solve the problem of high reagent cost, and achieve the effects of improved detection accuracy, large linear range, and simple formula

Inactive Publication Date: 2016-04-20
江苏迈源生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since lactic acid and ascorbic acid interfere with the reaction, an anti-interference agent needs to be added, and the reagent cost is relatively high

Method used

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  • Determining method and determining kit for beta-hydroxybutyrate
  • Determining method and determining kit for beta-hydroxybutyrate
  • Determining method and determining kit for beta-hydroxybutyrate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Determination method of β-hydroxybutyrate: β-hydroxybutyrate in the sample is detected by β-hydroxybutyrate dehydrogenase and thiooxidized coenzyme Ⅰ (Thio-NAD + ) specific oxidation to generate acetoacetate and thio-reduced coenzyme Ⅰ (Thio-NADH); acetoacetate generates β-hydroxybutyrate in the presence of β-hydroxybutyrate dehydrogenase and reduced coenzyme Ⅰ (NADH) and oxidized coenzyme I (NAD ＋ ), the reaction cycle is carried out, and a large amount of thio-reduced coenzyme Ⅰ (Thio-NADH) is produced, and the content of β-hydroxybutyric acid in the sample is quantified by measuring the absorbance change of the generated Thio-NADH at a wavelength of 405nm. A small amount of β-hydroxybutyric acid plays an amplifying role, and its reaction equation is as follows:

[0039]

[0040] The kit for the determination of β-hydroxybutyric acid is a double reagent, which is divided into reagent A and reagent B.

[0041] Reagent A:

[0042] Phosphate buffer (pH4.0) 5mmol / L ...

Embodiment 2

[0057] The assay method of β-hydroxybutyric acid is with embodiment 1;

[0058] The kit for the determination of β-hydroxybutyric acid is a double reagent, which is divided into reagent A and reagent B.

[0059] Reagent A:

[0060] MES buffer (pH4.0) 8mmol / L

[0061] Thio-NAD + 0.05mmol / L

[0062] TritonX-1001.0ml / L

[0063] Preservative NaN 3 0.5g / L

[0064] Reagent B:

[0065] Caps buffer (pH9.0) 200mmol / L

[0066] β-hydroxybutyrate dehydrogenase 3KU / L

[0067] NADH0.10mmol / L

[0068] Stabilizer ethylene glycol 70ml / L

[0069] Its detection method and calculation formula are the same as in Example 1.

Embodiment 3

[0071] The assay method of β-hydroxybutyric acid is with embodiment 1;

[0072] The reagents for the determination of β-hydroxybutyric acid are divided into reagent A and reagent B.

[0073] Reagent A:

[0074] Glycine buffer (pH4.5) 10mmol / L

[0075] Thio-NAD + 0.15mmol / L

[0076] Tween-20 (Tween-20) 0.5g / L

[0077] Preservative NaN 3 0.9g / L

[0078] Reagent B:

[0079] Tris buffer (pH9.0) 300mmol / L

[0080] β-hydroxybutyrate dehydrogenase 4KU / L

[0081] NADH0.08mmol / L

[0082] Bovine Serum Albumin (BSA) 1.0g / L

[0083] Ethylene glycol 300ml / L

[0084] Its detection method and calculation formula are the same as in Example 1.

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Abstract

The invention relates to a cyclophorase determining method and a determining kit for beta-hydroxybutyrate. According to the method, beta-hydroxybutyrate is specifically oxidized by beta-hydroxybutyrate dehydrogenase and Thio-NAD<+>, and acetoacetic acid and Thio-NADH are produced; beta-hydroxybutyrate and NAD<+> are produced by acetoacetic acid in presence of beta-hydroxybutyrate dehydrogenase and NADH; reactions circulate, a large amount of Thio-NADH is produced, the content of beta-hydroxybutyrate can be calculated through determination of the absorbance increase amount rate of the produced Thio-NADH under the condition that the wavelength is 405 nm, the method has an amplification action on trace beta-hydroxybutyrate and has the advantages of high sensitivity, good stability, low cost and the like. The kit comprises NADH, beta-hydroxybutyrate dehydrogenase, Thio-NAD<+>, a buffer solution, a surfactant and a stabilizer. The method can be used for determining the content of beta-hydroxybutyrate through a biochemical analyzer.

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 a circulating enzyme of β-hydroxybutyric acid and a detection kit thereof. Background technique [0002] Ketone bodies are metabolites of triglycerides. When the glucose in the body is absolutely or relatively insufficient, a large amount of triglycerides in the body will be decomposed to generate glycerol and fatty acids, and the fatty acids will be oxidized in the liver to generate energy and ketone bodies, which include 2% acetone, 23% acetoacetate and 75% β-hydroxybutyric acid, if the ketone body content in the body is too high, it will cause ketoacidosis. Ketotoxicity is a life-threatening condition. Timely and accurate detection of ketone body levels is a very necessary means to predict ketoacidosis. Ketone bodies are also an important marker for the detection and control of diabetes. [0003] Ketone bodies,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/3103
Inventor 孟广宇吉爽爽李丰盛
Owner 江苏迈源生物科技有限公司
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