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Method for quantitative determination of biochemical substance by enzyme analysis via predicting background

A technology for the quantitative determination of biochemical substances, applied in the field of clinical testing, can solve the problems of many interference factors, high cost, long reaction time, etc., and achieve the effect of high analysis efficiency, low cost and high accuracy

Inactive Publication Date: 2004-04-14
廖飞
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Kinetic methods can be used for indirect quantitative determination of products, but the cost is high and there are many interference factors; if the end-point balance method is used, it is very sensitive to the background and requires a long reaction time, so it cannot be automatically determined

Method used

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  • Method for quantitative determination of biochemical substance by enzyme analysis via predicting background
  • Method for quantitative determination of biochemical substance by enzyme analysis via predicting background
  • Method for quantitative determination of biochemical substance by enzyme analysis via predicting background

Examples

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Effect test

Embodiment 1

[0037] Example 1: see figure 1 , image 3 , Figure 4 and Figure 5 , carry out enzymatic analysis and measure the method for uric acid content in the urine sample by predicting background, it is characterized in that carry out as follows:

[0038] (1) Take 6 μl of urine sample and add it to the test tube;

[0039] (2), get 1.17ml, the concentration is 50mmol / L, and the boric acid buffer solution containing 50 μmol / L iron ion complexing agent diethyltriaminopentaacetic acid (DETAPA), pH9.2 is added in the same test tube;

[0040] (3), the mixed solution in the above-mentioned test tube is vortexed and fully mixed;

[0041] (4), the mixed solution in the above-mentioned test tube was kept at a constant temperature of 10 minutes in a water bath at 25°C;

[0042] (5) Add 24 μl of Cndida Utilis uricase solution with a concentration of 0.8-1.0U / ml in the test tube, or Bacillus fastidasus uricase; mix immediately, start timing with the moment of adding the enzyme as the startin...

Embodiment 2

[0057] Example 2: see figure 2 , image 3 , Figure 4 and Figure 5 A method for carrying out enzymatic analysis and determination of uric acid content in plasma samples by predicting the background, characterized in that it is carried out as follows:

[0058] (1) Take 20 μl of plasma sample and add it to the test tube;

[0059] (2), get 1.17ml, concentration is the boric acid buffer solution of 50mmol / L, containing 50μmol / L iron ion complexing agent diethyltriaminopentaacetic acid (DETAPA), pH9.2, add in the same test tube;

[0060] (3), the mixed solution in the above-mentioned test tube is vortexed and fully mixed;

[0061] (4), above-mentioned test tube is kept at constant temperature in 25 ℃ of water baths for 10 minutes;

[0062] (5) Add 10 μl of Cndida Utilis uricase solution with a concentration of 0.8-1.0U / ml in the test tube, or Bacillus fastidiosus uricase; mix immediately, and start timing with the moment of adding the enzyme as the starting point of the reacti...

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Abstract

The method for quantitative determination of biochemical substance by means of predetermining background and making enzyme analysis includes the following steps: adding buffer solution and tool enzyme in the sample solution to be determined, measuring the change of light absorption with reaction time, and utilizing integral velocity equation to fit and analyze the negative curve of light adsorption consumed by substrate to predetermine background instead of directly detemining background. It can directly determine body fluid sample, and it has no meed of diluting said sample, and its determination accuracy is high.

Description

technical field [0001] The invention relates to a method for determining the content of specific biochemical substances in body fluids or other mixtures in the field of clinical testing, especially a method for quantitatively determining biochemical substances through enzymatic analysis by predicting the background. Background technique [0002] Enzymatic analysis has high selectivity in determining the content of specific substances, and is of great value in the determination of specific biochemical substances in mixtures. There are two main types of enzymatic analysis methods for determining the content of specific biochemical substances in mixtures: kinetic methods and end-point equilibrium methods. The end-point equilibrium method requires high concentrations of analytical tool enzymes and a long reaction time to ensure that the substrate is completely converted into the product, and then the changes in the corresponding detection indicators are m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/58C12Q1/62
Inventor 廖飞
Owner 廖飞
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