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Method and reaction kit for quantitative detection of sarcosine content

A technology of reaction reagent and sarcosine, which is applied in the field of quantitative detection of sarcosine content, can solve the problem that the detection method is not suitable for the determination of sarcosine content at lower concentrations, and achieves the effects of low detection limit, high sensitivity and less interference

Active Publication Date: 2011-12-14
泰安泰笙医药科技有限公司
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0003] The purpose of the embodiments of the present invention is to provide a method for quantitatively detecting sarcosine content, aiming to solve the problem that existing detection methods are not suitable for the determination of sarcosine content at lower concentrations

Method used

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  • Method and reaction kit for quantitative detection of sarcosine content
  • Method and reaction kit for quantitative detection of sarcosine content
  • Method and reaction kit for quantitative detection of sarcosine content

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Embodiment 1

[0046] 1. a. Prepare 0.5ml concentration of 1.0×10 -6 mol / L sarcosine solution, the temperature is controlled at 30°C, and NH with pH=10.4 is added 3 -NH 4 Cl buffer solution (0.2mol / L) prepared 4.49ml 3.0×10 -5 mol / L p-cresol solution, 0.005ml sarcosine oxidase solution (10000units / L) and 0.005ml peroxidase solution (20000units / L), mix homogeneously (in the final reaction mixture, the actual concentration of sarcosine is 0.1×10 -6 mol / L), after reacting for 20 minutes, measure the fluorescence emission spectrum of the system with a fluorescence spectrophotometer (optimum excitation wavelength λex320nm, emission spectrum scanning range 350-550nm). For its fluorescence emission spectrum see figure 2 Medium Cl.

[0047] b. Prepare 0.5ml concentration of 1.0×10 -5 mol / L sarcosine solution, other conditions are the same as a. The actual concentration of sarcosine in the final reaction mixture was 1.0×10 -6 mol / L, its fluorescence emission spectrum see figure 2 Middle C2...

Embodiment 2

[0055] The pH value of the solution used is 9, and the working reagent is mixed according to the following concentration requirements (p-methylphenol, 1×10 -7 mol / L; sarcosine oxidase, 10units / L; peroxidase, 20units / L; sarcosine from urine samples, reacted for 20 minutes to completely react sarcosine and hydrogen peroxide, and use a fluorescence spectrometer Measure the fluorescence excitation spectrum to obtain the fluorescence intensity. Calculate the concentration of sarcosine from the relationship curve of sarcosine concentration-fluorescence intensity to be 0.1×10 -9 mol / L.

Embodiment 3

[0057] The pH value of the solution used is 10.4, and the working reagent is mixed according to the following concentration requirements (p-cresol, 1×10 -5 mol / L; sarcosine oxidase, 10units / L; peroxidase, 20units / L; sarcosine from urine sample), react for 20 minutes to make sarcosine and hydrogen peroxide react completely, and use fluorescence The spectrometer measures the fluorescence excitation spectrum to obtain the fluorescence intensity. The sarcosine concentration obtained from the sarcosine concentration-fluorescence intensity relationship curve is 5.43×10 -7 mol / L.

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Abstract

The invention provides a method for quantitatively detecting sarcosine content and a reaction kit, the method comprising steps: S1 preparing sarcosine solutions with different concentrations, adding a reaction reagent to the solution to completely react sarcosine and hydrogen peroxide ; S2 measures the fluorescence emission spectrum of the fluorescent dimer of the reaction product in S1 respectively, and draws the sarcosine concentration-fluorescence intensity relationship curve; S3 uses the method as described in S1 to make the unknown sarcosine sample occur as described in S1 For the reaction, measure the fluorescence emission spectrum of the reaction product to obtain the fluorescence intensity data, and compare the sarcosine concentration-fluorescence intensity relationship curve to obtain the content of the unknown sarcosine sample. The reaction kit components of the present invention include p-cresol or p-hydroxyphenylacetic acid solution, sarcosine oxidase, peroxidase, and alkaline buffer. The method of the invention is very suitable for mass analysis and detection of trace amounts of sarcosine in urine samples or blood samples.

Description

technical field [0001] The invention relates to a method for quantitatively detecting sarcosine content and a reaction kit, in particular to a method for quantitatively detecting sarcosine content by an enzyme-catalyzed fluorescence method, and a reaction kit used in the method for detecting sarcosine content. Background technique [0002] At present, the determination of sarcosine content is generally carried out by liquid-gas chromatography coupled with mass spectrometry. The instruments used in this method are expensive and the operation is complicated, which is not convenient for large-scale application. In addition, a spectrophotometric method for the determination of creatinine in blood and urine by an enzymatic method in a clinical laboratory uses creatinine to react with enzymes such as creatinine aminohydrolase, creatine amidinohydrolase, sarcosine oxidase, and peroxidase and water. The chromogen substance is generated under the joint action of chromogen, oxygen and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 杨海朋戈早川
Owner 泰安泰笙医药科技有限公司
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