Screening method for xanthine oxidase inhibitors and/or superoxide anion scavengers

A technology of xanthine oxidase and superoxide anion is applied in the field of screening xanthine oxidase inhibitors and/or superoxide anion scavengers, and achieves the effects of simple operation, low experimental cost, easy identification and memory

Inactive Publication Date: 2015-08-19
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional screening methods for xanthine oxidase inhibitory activity or superoxide anion scavenging activity require compounds to be isolated and purified from traditional Chinese medicines and require a large enough amount to evaluate the activity of medicinal samples through various experimental methods in vivo and in vitro, but many Natural compounds obtained from traditional Chinese medicine are difficult to meet these conditions

Method used

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  • Screening method for xanthine oxidase inhibitors and/or superoxide anion scavengers
  • Screening method for xanthine oxidase inhibitors and/or superoxide anion scavengers
  • Screening method for xanthine oxidase inhibitors and/or superoxide anion scavengers

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Embodiment 1

[0024] 1.1 Drugs and reagents

[0025] Psoralen decoction pieces were purchased from Shanghai Kangqiao Decoction Pieces Factory, xanthine oxidase (X4875-10UN, LOT#110M7004V, purchased from sigma), xanthine (X0625-5G, lot number 062K5306, purchased from sigma), nitrotetrazole chloride Nitrogen blue (NBT) (N6876-5G, lot number 97569LJ, purchased from sigma), potassium dihydrogen phosphate (lot number F20100202, purchased from Sinopharm Chemical Reagent Co., Ltd.), dipotassium hydrogen phosphate (lot number F20100118, purchased from Sinopharm Chemical Reagent Co., Ltd.) Co., Ltd.), sodium hydroxide (batch number F20100926, purchased from Sinopharm Group Chemical Reagent Co., Ltd.), disodium ethylenediaminetetraacetic acid (EDTA) (batch number F20080616, purchased from Sinopharm Group Chemical Reagent Co., Ltd.).

[0026] 1.2 Instruments

[0027]Super constant temperature bath (Shanghai Bilang Instrument Co., Ltd.), electronic analytical balance (BP211D, Sartorius, Germany), elec...

Embodiment 2

[0038] 2.1 Drugs and reagents

[0039] Allopurinol (purchased from sigma), xanthine oxidase (X4875-10UN, LOT #110M7004V, purchased from sigma) xanthine (X0625-5G, lot number 062K5306, purchased from sigma), nitroblue tetrazolium chloride ( NBT) (N6876-5G, batch number 97569LJ, purchased from sigma), potassium dihydrogen phosphate (batch number F20100202, purchased from Sinopharm Chemical Reagent Co., Ltd.), dipotassium hydrogen phosphate (batch number F20100118, purchased from Sinopharm Chemical Reagent Co., Ltd.) , sodium hydroxide (batch number F20100926, purchased from Sinopharm Chemical Reagent Co., Ltd.), disodium ethylenediaminetetraacetic acid (EDTA) (batch number F20080616, purchased from Sinopharm Chemical Reagent Co., Ltd.).

[0040] 2.2 Instruments

[0041] Super constant temperature bath (Shanghai Bilang Instrument Co., Ltd.), electronic analytical balance (BP211D, Sartorius, Germany), electric thermostatic blast drying oven (DHG-9053A type, Shanghai Yiheng Techno...

Embodiment 3

[0051] 3.1 Drugs and reagents

[0052] Ursolic acid standard, rosmarinic acid standard, quercetin standard (all purchased from Shanghai Center for Traditional Chinese Medicine Standardization Research), allopurinol (purchased from sigma), xanthine oxidase (X4875-10UN, LOT#110M7004V , purchased from sigma), xanthine (X0625-5G, batch number 062K5306, purchased from sigma), nitroblue tetrazolium chloride (NBT) (N6876-5G, batch number 97569LJ, purchased from sigma), potassium dihydrogen phosphate ( Batch number F20100202, purchased from Sinopharm Chemical Reagent Co., Ltd.), dipotassium hydrogen phosphate (batch number F20100118, purchased from Sinopharm Chemical Reagent Co., Ltd.), sodium hydroxide (batch number F20100926, purchased from Sinopharm Chemical Reagent Co., Ltd.), B2 Disodium amine tetraacetate (EDTA) (batch number F20080616, purchased from Sinopharm Chemical Reagent Co., Ltd.).

[0053] 3.2 Instruments

[0054] Super constant temperature bath (Shanghai Bilang Instr...

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Abstract

The invention discloses a method for fast screening a xanthine oxidase inhibitor or a superoxide anion scavenging agent by utilizing a coupling technique of thin-layer chromatography and bioautography. According to the method, the thin-layer chromatography is combined with biological activity assay, a superoxide anion free radical (O2<.->) generated by an enzymatic reaction reacts with nitroblue tetrazolium (NBT) to generate formazan which is a bluish violet insoluble substance, and substances capable of inhibiting xanthine oxidase or scavenging superoxide anion inhibit the reaction so yellow and / or white spots under a purple background show on a thin layer plate. The method is suitable for analyzing the in-vitro inhibition activity to the the xanthine oxidase or the scavenging activity to the superoxide anion of natural drugs and / or traditional Chinese medicine extracts or monomeric compounds.

Description

technical field [0001] The invention relates to a method for screening xanthine oxidase inhibitors and / or superoxide anion scavengers by using xanthine oxidase-thin-layer chromatography bioautographic technology. Background technique [0002] Xanthine oxidase is an important enzyme in nucleic acid metabolism in the body, and is widely distributed in the cytoplasm of human heart, lung, liver and other tissues. Xanthine oxidase can catalyze the oxidation of xanthine and hypoxanthine to produce uric acid and produce peroxide free radicals. Excessive uric acid concentration can lead to hyperuricemia and can trigger gout attacks. Gout is a disease that crystallizes and precipitates in joints due to excessive concentration of uric acid in tissue and body fluids, causing joint pain. It is mainly caused by abnormal generation of uric acid and superoxide anion due to purine metabolism disorder. One of the chronic diseases. The latest epidemiological research shows that although th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/90
Inventor 吴弢李相坤唐吉鹤陶宏迅孙天杰
Owner SHANGHAI UNIV OF T C M
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