Method for quantitative determination content of sarcosine and reaction kits

A reaction reagent and quantitative detection technology, which is applied in the field of quantitative detection of sarcosine content, can solve the problems that the detection method is not applicable to the determination of low concentration sarcosine content, and achieve the effects of low detection limit, good repeatability and low cost

Active Publication Date: 2009-11-25
泰安泰笙医药科技有限公司
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  • Abstract
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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 for quantitative determination content of sarcosine and reaction kits
  • Method for quantitative determination content of sarcosine and reaction kits
  • Method for quantitative determination content of sarcosine and reaction kits

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Experimental program
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Embodiment 1

[0065] 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 λex 320nm, emission spectrum scanning range 350-550nm). For its fluorescence emission spectrum see figure 2 Middle C1.

[0066] 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 C...

Embodiment 2

[0074] 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

[0076] 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 quantitative determination content of sarcosine and reaction kits, the method includes steps: S1, preparing sarcosine solution of different concentration, adding reaction agent to the solution to react sarcosine with peroxide completely; S2, detecting fluorescence spectra of products fluorescent dimer in S1 respectively, drawing relationship curve of sarcosine content-flurescence; S3, using the method of S1 to make sarcosine samples react as described in S1, measuring fluorescence emission spectra of reaction product to obtain fluorescence data, then comparing with the relationship curve of sarcosine content-flurescence to obtain content of unknown sarcosine samples. Components of the reaction kits according to the invention includes methyl phenol or P-hydroxy acid solution, sarcosine oxidase, peroxydase and alkaline buffer. Method according to the invention is very suitable for large-scale analytica test of minim sarcosine in urine sample or blood sample.

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 Applications(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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