Enzymatic activity analysis method for urate oxidase

A uric acid oxidase and analysis method technology, applied in the biological field, can solve problems such as linear relationships, and achieve the effect of reducing errors

Inactive Publication Date: 2006-10-04
吉林修正药业新药开发有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0009] The above method uses 20% KOH to terminate the enzyme reaction, and the absor

Method used

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  • Enzymatic activity analysis method for urate oxidase
  • Enzymatic activity analysis method for urate oxidase

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Basic principle of measurement

[0025] Uric acid oxidase can specifically oxidize uric acid to allantoin under certain conditions in vitro, and the decrease in the concentration of uric acid under the oxidative decomposition of urate oxidase reflects the level of enzyme activity. This method is based on urate oxidase oxidizing uric acid to allantoin. The maximum absorption wavelength of uric acid is 292nm, and the maximum absorption wavelength of oxidized product allantoin is 224nm. The maximum absorption wavelengths of the two are different, and the absorption value of uric acid at 292nm is proportional to its concentration within a certain range. , Quantitative determination of uric acid can be carried out by spectrophotometry. Select the appropriate enzyme and substrate uric acid concentration, pH 8.9, react at 30°C for a certain period of time, use the 292nm absorbance of different concentrations of uric acid standards to develop a standard curve, and calculate ...

Embodiment 2

[0059] Enzyme reaction volume 5ml, buffer solution TEA (7.5g triethanolamine; 0.38g EDTA), pH8.9, uric acid 60μmol / L, keep warm at 30°C, reaction time 5min, stop the reaction with 20% KOH and 3.5mol / L sulfuric acid respectively, The detection wavelength is 292nm. Concrete determination method is as embodiment 1. Determination at different times, the results are shown in Table 1.

[0060]

Embodiment 3

[0062] Enzyme reaction volume 5ml, buffer solution TEA (7.5g triethanolamine; 0.38g EDTA), pH 10, uric acid 180μmol / L, keep warm at 35°C, reaction time 25min, stop reaction with 9mol / L sulfuric acid, detection wavelength 292nm. Concrete determination method is as embodiment 1.

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Abstract

The related activity analysis method for urate oxidase comprises: in internal analysis, injecting urate oxidase into the animal contained no the urate oxidase and then collecting serum; in external analysis, with enzyme reaction, stopping acid liquid, taking analysis with ultraviolet spectrophotometry. This invention can reduces error greatly.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to an activity analysis method of urate oxidase. technical background [0002] Uric acid is the final product of the metabolism of purine compounds in the human body, and the blood uric acid level depends on the dynamic balance between uric acid production and excretion. Increased uric acid production or decreased uric acid excretion can lead to elevated blood urate concentrations. Plasma uric acid saturation: 380-420 μmol / L (6.4-7.1 mg / dL) at 37°C and pH 7.4. [0003] Clinically, elevated blood uric acid levels are related to the following conditions: (1) Leukemia and other malignant tumors, due to the rapid proliferation of malignant cells and enhanced nucleic acid decomposition, blood uric acid increases. The increase in blood uric acid was more obvious after tumor chemotherapy. Patients with eclampsia can also be elevated. (2) Gout is caused by th...

Claims

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Application Information

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IPC IPC(8): C12Q1/62C12Q1/26G01N33/96
Inventor 刘国安刘沐荣康经武
Owner 吉林修正药业新药开发有限公司
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