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Kit for detecting D-3-hydroxybutyric acid and preparation method of kit

A D-3-, reagent kit technology, applied in the field of medicine and biochemistry, can solve the problems of radioactive contamination by radiochemical method, complex operation of gas chromatography, poor stability of reagents, etc., and achieve simple operation and short storage time , Improve the effect of measurement accuracy

Inactive Publication Date: 2016-08-10
ANHUI IPROCOM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a method for the determination of D-3-hydroxybutane Acid kit and preparation method thereof

Method used

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  • Kit for detecting D-3-hydroxybutyric acid and preparation method of kit
  • Kit for detecting D-3-hydroxybutyric acid and preparation method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0057] Reagent R1:

[0058] Tris Buffer 100mmol / L

[0059] D-3-hydroxybutyrate dehydrogenase 2.5KU / L

[0060] Polyoxypropylene polyoxyethylene copolymer 3.0mL / L

[0061] Sodium azide 0.035 mmol / L

[0062] Its solvent is purified water.

[0063] Reagent R2:

[0064] Oxalate 25mmol / L

[0065] NAD+ 2.5mmol / L

[0066] Sodium azide 0.035 mmol / L

[0067] Its solvent is purified water.

Embodiment 2

[0069] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0070] Reagent R1:

[0071] 4-Hydroxyethylpiperazineethanesulfonic acid buffer 90mmol / L

[0072] D-3-hydroxybutyrate dehydrogenase 3.1KU / L

[0073] Polyoxypropylene polyoxyethylene copolymer 5.0mL / L

[0074] Sodium azide 0.059 mmol / L

[0075] Its solvent is purified water.

[0076] Reagent R2:

[0077]Oxalate 20mmol / L

[0078] NAD+ 3.0mmol / L

[0079] Sodium azide 0.059 mmol / L

[0080] Its solvent is purified water.

Embodiment 3

[0082] Kit preparation and method of use

[0083] 1. Prepare the reagent according to the content of the following components:

[0084] Reagent R1:

[0085] Tris Buffer 100mmol / L

[0086] D-3-hydroxybutyrate dehydrogenase 2.5KU / L

[0087] Polyoxypropylene polyoxyethylene copolymer 3.0mL / L

[0088] Sodium azide 0.035 mmol / L

[0089] Its solvent is purified water.

[0090] Reagent R2:

[0091] Oxalate 25mmol / L

[0092] NAD+ 2.5mmol / L

[0093] Sodium azide 0.035 mmol / L

[0094] Its solvent is purified water.

[0095] 2. Parameter setting of automatic biochemical analyzer

[0096] (a) Detection temperature: 37°C;

[0097] (b) Detection wavelength: main wavelength 340nm, secondary wavelength 700nm;

[0098] (c) Reaction time: 8 minutes, among them, the incubation time is 5 minutes, measure and read the absorbance A1 immediately after adding the reagent R2, read the absorbance A2 after 3 minutes, and calculate the change of absorbance ΔA = A2-A1;

[0099] (d) Reaction...

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Abstract

The invention discloses a kit for detecting D-3-hydroxybutyric acid and a preparation method of the kit .The kit comprises a kit body composed of two independent liquid components, namely a reagent R1 and a reagent R2 .The reagent R1 is prepared from 20-180 mmol / L of buffer solution, 0.1-4.9 KU / L of D-3-hydroxybutyric acid dehydrogenase, 0.1-6.0 mL / L of polypropylene oxide and polyoxyethylene copolymer and 0.01-0.059 mmol / L of preservative, wherein solvent is purified water; the reagent R2 is prepared from 10-40 mmol / L of oxalate, 0.1-4.9 mmol / L of NAD+ and 0.01-0.059 mmol / L of preservative, wherein solvent is purified water .The invention further discloses the preparation and use method of the kit for detecting D-3-hydroxybutyric acid .The preparation and use method includes the following steps of preparing the reagents according to the contents of the components, mixing the reagent R1 and the reagent R2 with a to-be-detected sample to react sufficiently, detecting the after-reaction light absorption degree differential value through a full-automatic biochemical analyzer, and calculating the concentration of D-3-hydroxybutyric acid in the sample according to the light absorption degree change value .The kit has the advantages of being free of pollution, high in stability and the like.

Description

technical field [0001] The invention relates to the technical fields of medicine and biochemistry, in particular to a kit for measuring D-3-hydroxybutyric acid and a preparation method thereof. Background technique [0002] Diabetes is a common and frequently-occurring disease, and its incidence is increasing year by year. When the metabolic disorder of the body develops to accelerate the decomposition of fat and the accumulation of serum ketones exceeds the normal level, it is called ketosis, and its clinical manifestation is ketosis; when metabolic acidosis occurs due to the accumulation of ketoacids, it is called ketoacidosis, such as When the condition is so severe that coma occurs, it becomes "diabetic coma". The main component of ketone bodies is D-3-hydroxybutyric acid, accounting for about 78% of the total ketone bodies. It is the main substance of diabetic ketoacidosis. It lacks typical clinical symptoms in the early stage, while the traditional ketone body detec...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N21/31
CPCG01N33/52G01N21/31
Inventor 蔡晓辉庄庆华吴铮徐运
Owner ANHUI IPROCOM BIOTECH CO LTD
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