Method for rapidly screening active ingredients of drugs based on ultrafiltration-liquid chromatography-mass spectrometry

A liquid chromatography and drug screening technology, which is applied in the field of analysis of natural active ingredients of traditional Chinese medicine and drug synthesis, can solve the problems of rapid screening of drug active ingredients, slow screening of active ingredients, and low content of medicinal substances. Achieve the effect of fast batch screening, simple operation, and less loss

Inactive Publication Date: 2019-04-09
XIAN UNIV OF SCI & TECH
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Problems solved by technology

However, the traditional research mode of "component separation-structure identification-activity screening" has the disadvantages of heavy workload, low separation efficiency, and slow screening of active components; at the same time, the characteristics of complex components and relatively low content of medicinal substances increase the The research difficulty of a model
[0006] Ultrafiltration-UPLC-MS technology is currently usually used for the detection of free target substances in samples, and there is no report on the use of it for rapid screening of active pharmaceutical ingredients

Method used

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  • Method for rapidly screening active ingredients of drugs based on ultrafiltration-liquid chromatography-mass spectrometry

Examples

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Effect test

example 1

[0032] Rapid Screening of PTP1B Inhibitors in Example 1 Astragalus Extract

[0033] Astragalus is widely used clinically, and it is the main drug in hypoglycemic prescriptions such as Jinqi Jiangtang Tablets and Huangqi Liuyi Decoction. In addition, Astragalus Injection also has a good effect on the treatment of diabetes and diabetic nephropathy.

[0034] (1) Co-incubation of Astragalus extract and PTP1B enzyme

[0035] Astragalus extract (75% ethanol extract, Xi’an Tianrui Biological Company, 500 μg / mL) 20 μL, PTP1B enzyme (hGST-PTP1 50 nmol / L) 20 μL, and ammonium acetate buffer solution (pH=5.2) 100 μL were mixed in an EP tube , to obtain a combined sample, and then place the EP tube in a 37°C water bath and incubate with vibration for 30 min. After incubation, transfer to an ultrafiltration tube (PALL Nanosep centrifugal concentration tube 10K) and centrifuge at 10000g for 10min. Continue to add 100 μL of ammonium acetate buffer solution (pH=5.2) to the insoluble matter i...

example 2

[0045] Quantitative experiment of puerarin and α-glucosidase in example 2, kudzu root

[0046] Puerarin, also known as puerarin, is an isoflavone compound isolated from Pueraria lobata, a traditional Chinese medicine. It exists in the roots of leguminous plants Pueraria lobata and Pueraria mirifica. Early reports have antipyretic, sedative and coronary blood flow increased effect. Recent pharmacological studies have shown that puerarin also has the effect of lowering blood sugar, and its target may be α-glucosidase.

[0047] (1) Co-incubation of kudzu root extract and α-glucosidase

[0048] Pueraria root extract (Xi'an Aiwo Biotechnology Co., Ltd., 1mg / mL, puerarin content greater than 6%) 20μL, α-glucosidase (0.2u / mL) 20μL, and ammonium acetate buffer solution (pH=5.2) 100μL mixed In the EP tube, the combined sample was obtained, and then the EP tube was placed in a 37°C water bath and incubated with shaking for 30 min. After incubation, transfer to an ultrafiltration tube...

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Abstract

The invention discloses a method for rapidly screening active ingredients of drugs based on an ultrafiltration-liquid chromatography-mass spectrometry. The method comprises the steps of dissolving a small molecular mixture of traditional Chinese medicine extract or other sources in a buffer solution, and adding enzyme for co-incubation; filtering the solution by ultracentrifugation concentration,repeating the filtering for multiple times, merging concentrated solutions, and making up the volume by the buffer solution; and analyzing the change of a chromatographic peak after the co-incubationof the enzyme by adopting a liquid chromatography-mass spectrometry technology. The method disclosed by the invention has the advantages that the hypoglycemic activity ingredients of the traditional Chinese medicines are screened by virtue of the ultrafiltration-UPLC-MS; and an experiment result shows that the method can carry out preliminary rapid screening on active small molecule compounds of anatural source in the complex ingredients. The method has the characteristics of simple operation, rapid batch screening, good experiment repeatability, visual and reliable experiment data and the like; and a new idea and method is provided for pharmacodynamic material basis research of the traditional Chinese medicines or the compounds.

Description

technical field [0001] The invention belongs to the field of analysis of natural active components of traditional Chinese medicine and synthesis of drugs, and relates to a method for rapidly screening active components of drugs based on ultrafiltration (Ultrafiltration)-HPLC / UPLC-MS technology. Background technique [0002] Diabetes is a common chronic disease in modern society. Its main clinical manifestations are hyperglycemia and disorders of sugar, protein, fat, water and electrolytes. According to the theory of traditional Chinese medicine, most of the diseases belong to "diabetes", while modern medicine divides diabetes into type I and type II. The pathogenesis of diabetes is complicated, and the advantages of traditional Chinese medicine in lowering blood sugar are mainly reflected in the multi-target effect. While lowering blood sugar, it also takes into account lowering lipid and anti-oxidation. Therefore, in the treatment of common chronic diseases such as diabete...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/047
Inventor 陈福欣严贝贝陈苏英杨倩倩白巧秀杨雅雯爱森·瓦卡斯周安宁龚频
Owner XIAN UNIV OF SCI & TECH
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