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Method for screening xanthine oxidase inhibitor

A technology of xanthine oxidase and screening method, which is applied in the field of screening of xanthine oxidase inhibitors, can solve problems such as inability to apply complex system specificity, and achieve the effects of low experimental cost, simple operation, and improved specificity

Active Publication Date: 2020-11-06
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to overcome the defects that the prior art cannot be applied to complex systems and has poor specificity, and provides a method for screening xanthine oxidase inhibitors using xanthine oxidase-thin-layer chromatography bioautographic technology with strong specificity

Method used

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  • Method for screening xanthine oxidase inhibitor
  • Method for screening xanthine oxidase inhibitor
  • Method for screening xanthine oxidase inhibitor

Examples

Experimental program
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Effect test

Embodiment 1

[0047] A screening method for xanthine oxidase inhibitors, the specific operations are as follows:

[0048] (1) The preparation process of various solutions is as follows:

[0049] Preparation of developer solution: Weigh 20g of sodium tungstate, dissolve it in 150mL of distilled water, add 16mL of phosphoric acid solution, heat and reflux for 2 hours, cool to room temperature, and dilute to 280mL with distilled water to make phosphotungstic acid storage solution, keep away from light For storage, dilute 1.45 times with distilled water before use;

[0050] Preparation of phosphate buffer: Accurately weigh 22.822g of dipotassium hydrogen phosphate (analytical pure), 13.609g of potassium dihydrogen phosphate (analytical pure), add them to a 500mL volumetric flask, and distilled water to make up the phosphoric acid of pH=7.1 buffer;

[0051] Preparation of xanthine oxidase solution: Dissolve EDTA disodium salt with pH=7.1 phosphate buffer solution, and finally prepare enzymatic...

Embodiment 2

[0060] A screening method for xanthine oxidase inhibitors, the specific operations are as follows:

[0061] (1) The preparation process of various solutions is as follows:

[0062] Preparation of developer solution: Weigh 20g of sodium tungstate, dissolve it in 150mL of distilled water, add 16mL of phosphoric acid solution, heat and reflux for 3 hours, cool to room temperature, and distill the volume to 280mL with distilled water to make phosphotungstic acid storage solution, keep away from light Store, dilute 1.6 times with distilled water before use;

[0063] Preparation of phosphate buffer solution: Accurately weigh 10.806g of dipotassium hydrogen phosphate (analytical pure), 2.245g of potassium dihydrogen phosphate (analytical pure), add them to a 500mL volumetric flask, and distilled water to make up the phosphoric acid of pH=7.6 buffer;

[0064] Preparation of xanthine oxidase solution: Dissolve EDTA disodium salt with pH=7.6 phosphate buffer solution, and finally prep...

Embodiment 3

[0072] A screening method for xanthine oxidase inhibitors, the specific operations are as follows:

[0073] (1) The preparation process of various solutions is as follows:

[0074] Preparation of developer solution: Weigh 20g of sodium tungstate, dissolve it in 150mL of distilled water, add 16mL of phosphoric acid solution, heat and reflux for 4 hours, cool to room temperature, and distill the volume to 280mL with distilled water to make phosphotungstic acid storage solution, keep away from light For storage, dilute 1.75 times with distilled water before use;

[0075] Preparation of phosphate buffer solution: accurately weigh 6.008g of dipotassium hydrogen phosphate (analytical pure), 3.682g of potassium dihydrogen phosphate (analytical pure), add them to a 500ml volumetric flask, distill to volume, and prepare pH=8.0 Phosphate buffer;

[0076]Preparation of xanthine oxidase solution: Dissolve EDTA disodium salt in phosphate buffer solution with pH=8.0, and finally prepare e...

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Abstract

The invention discloses a method for screening a xanthine oxidase inhibitor. The method comprises the following steps: preparing a test solution, dispensing the test solution on a thin-layer plate, and / or after carrying out developing by using a developing solvent, volatilizing the solvent; soaking the thin-layer plate prepared in the previous step in a xanthine oxidase solution, taking out the thin-layer plate, and activating xanthine oxidase on the thin-layer plate; sequentially soaking the thin-layer plate prepared in the previous step in the xanthine solution and a Na2CO3 solution, takingthe thin-layer plate out after each time of soaking and placing the thin-layer plate at the temperature of 25-40 DEG C for reaction, and making the thin-layer plate soaked in a color developing agentsolution for a color developing reaction after the reaction is completed, wherein the color developing agent solution is a phosphotungstic acid solution. The screening method of the xanthine oxidase inhibitor is simple to operate and low in experiment cost, and does not need complex instruments and equipment; a result is provided in a color image form, is vivid and visual and is easy to identify and memorize; and the specificity is high, and the application prospect is wide.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis, and relates to a screening method for xanthine oxidase inhibitors, in particular to a screening method for xanthine oxidase inhibitors by using xanthine oxidase-thin-layer chromatography bioautographic technology. Background technique [0002] Gout is the most common inflammatory arthritis caused by deposition of monosodium urate (MSU) in the joints of chronic hyperuricemia. In addition to joint pain, gout and hyperuricemia are also associated with hypertension, diabetes, metabolic syndrome, and renal and cardiovascular diseases, causing great suffering to patients. Therefore, the treatment of gout is of great significance for alleviating the suffering of patients and improving the quality of life of patients. [0003] Uric acid-lowering therapy (ULT) is the basis for the treatment of gout. When the serum urate concentration is reduced to 6 mg / dL or below, it can effectively reduce ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/90G01N21/78
CPCG01N30/90G01N21/78
Inventor 吴弢苗雨
Owner SHANGHAI UNIV OF T C M
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