Liquid stable kit for measuring beta-hydroxybutyric acid by cyclic enzyme method

A cyclic enzymatic method and kit technology, which is applied in the field of medical testing and determination, can solve the problems of poor solubility of oxidized INT, limitations in large-scale promotion and application, and poor reagent stability, and achieves easy popularization and application, low cost, and simple method. Effect

Active Publication Date: 2012-05-02
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the poor solubility and instability of the oxidized INT in the reagents of the cyclic enzyme colorimetric method, most of the domestic reagent companies currently use the three-reagent method, that is, the solution containing nitro blue tetrazolium is mixed into the reagent 1 before use, and the mixed The solution can only be stable for 30 days at 2-8°C. In contrast, the operation steps are complicated and the stability of the reagent is far less than that of the kinetic method.
At the same time, because the determination is easily affected by lactic acid and ascorbic acid treatment, its large-scale clinical application is limited.

Method used

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  • Liquid stable kit for measuring beta-hydroxybutyric acid by cyclic enzyme method
  • Liquid stable kit for measuring beta-hydroxybutyric acid by cyclic enzyme method
  • Liquid stable kit for measuring beta-hydroxybutyric acid by cyclic enzyme method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Reagent 1: Tris-HCl (pH 9.0) 50mmol / L

[0038] β-Hydroxybutyrate dehydrogenase 1KU / L

[0039] Diaphorase 1KU / L

[0040] Tween 20 10g / L

[0041] BSA 10mmol / L

[0042] EDTA-2Na 20g / L

[0043] Glutathione 1g / L

[0044] Proclin 300 1ml / L

[0045] Reagent 2: citric acid-trisodium citrate (pH 4.5) 50mmol / L

[0046] Coenzyme I 2.5mmol / L

[0047] Nitro blue tetrazolium 1mmol / L

[0048] Mannitol 10mmol / L

[0049] Proclin 300 2ml / L

[0050] The preparation method of Reagent 1 and Reagent 2 is a conventional method, that is, the components of Reagent 1 and Reagent 2 are added to distilled water and then mixed and stirred evenly.

Embodiment 2

[0052]Reagent 1: Triethanolamine buffer (pH 7.0) 100mmol / L

[0053] β-hydroxybutyrate dehydrogenase 2.5KU / L

[0054] Diaphorase 2.5KU / L

[0055] Triton X-100 1g / L

[0056] Glycerol 20mmol / L

[0057] EDTA-2Na 20g / L

[0058] Vitamin C oxidase 40g / L

[0059] Proclin 300 5ml / L

[0060] Reagent 2: acetic acid-sodium acetate (pH 4.0) 100mmol / L

[0061] Coenzyme I 10mmol / L

[0062] Nitro blue tetrazolium 5mmol / L

[0063] β-cyclodextrin 10mmol / L

[0064] Proclin 300 3ml / L

[0065] The preparation method of the kit in Example 2 is the same as in Example 1.

Embodiment 3

[0067] Reagent 1: GOOD’S (pH 8.5) 200mmol / L

[0068] β-Hydroxybutyrate dehydrogenase 5KU / L

[0069] Diaphorase 5KU / L

[0070] Tween 80 5g / L

[0071] Glycerin 10mmol / L

[0072] Oxalic acid 20g / L

[0073] Vitamin C oxidase 100g / L

[0074] Proclin 300 1ml / L

[0075] Reagent 2: glycine buffer (pH 3.5) 200mmol / L

[0076] Coenzyme I 20mmol / L

[0077] Nitro blue tetrazolium 10mmol / L

[0078] Mannitol 10mmol / L

[0079] Proclin 300 1ml / L

[0080] The preparation method of the kit of Example 3 is the same as that of Example 1.

[0081] The test conditions for the determination of β-hydroxybutyric acid in the sample by using the kit of the embodiment of the present invention are as follows: temperature: 30-37° C.; optical path of the cuvette: 1.0 cm. The main detection wavelength is 505nm, and the secondary wavelength is 700nm.

[0082] The method of measuring the total bilirubin in the sample by using the β-hydroxybutyric acid assay kit of the embodiment of the present inven...

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Abstract

The invention discloses a liquid stable kit for measuring beta-hydroxybutyric acid by a cyclic enzyme method. The liquid stable kit consists of a reagent 1 and a reagent 2, wherein 1L of reagent 1 comprises 50 to 500mmol of buffer solution, 1 to 5KU of beta-hydroxybutyrate dehydrogenase, 1 to 5KU of diaphorase, 0.1 to 100g of surfactant, 1 to 100mmol of stabilizer, 0.1 to 100g of anti-interference agent I, 0.1 to 100g of anti-interference agent II, and 0.1 to 100ml of preservative; and 1L of reagent 2 comprises 50 to 500mmol of buffer solution, 1 to 20mmol of coenzyme I, 0.1 to 10mmol of nitrotetrazolium blue, 1 to 100mmol of stabilizer and 0.1 to 100ml of preservative. The liquid stable kit has the advantages of stability, wide linear range, high measurement accuracy, high antijamming capability and low cost.

Description

technical field [0001] The invention belongs to the technical field of medical testing and determination, and in particular relates to a liquid stable kit for measuring beta-hydroxybutyric acid by a cyclic enzyme method. Background technique [0002] Ketone bodies are the product of fat metabolism. Every cell in a normal human body contains fatty acids. When the body needs more energy, such as exceeding the energy that can be provided by glucose produced by intake of food, fat cells are required to release fatty acids into the blood. The fatty acids are then transported to the liver where they are converted into ketone bodies. The ketone bodies include acetone, acetoacetate and β-hydroxybutyrate, which are in dynamic equilibrium. Among them, β-hydroxybutyric acid has the most content, which can account for 75% of the total ketone bodies. Ketone bodies, like blood glucose, are an important marker of metabolic control. One of the most important characteristics of patients w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/64G01N21/31
Inventor 邹炳德沃燕波
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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