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A method for high-throughput screening of bioactive components in complex components by planar chromatography component microarray and its application

A technology of biologically active components and planar chromatography, applied in the field of biologically active components, can solve the problems of difficulty in implementation and difficulty in determining the chemical components of biological activity, and achieve the effect of ensuring scalability

Active Publication Date: 2022-02-18
GUANGDONG MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to make each chromatographic spot fall into the same microwell pool as much as possible, this method requires each plane chromatographic spot to be strictly corresponding to the microwell plate array unit, which is difficult to implement for thin-layer chromatograms with irregular spot shapes and positions Large; when a chromatographic spot falls into multiple microporous pools, and there are multiple co-existing components in the microporous pool, it is difficult to determine the chemical composition of the biological activity

Method used

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  • A method for high-throughput screening of bioactive components in complex components by planar chromatography component microarray and its application
  • A method for high-throughput screening of bioactive components in complex components by planar chromatography component microarray and its application
  • A method for high-throughput screening of bioactive components in complex components by planar chromatography component microarray and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Construct TLC fingerprints that fully represent the chemical composition of traditional Chinese medicine and natural product samples and reflect the differences and characteristics of the chemical composition of the samples

[0100] 1) Take 60 meshes of galangal dry powder as a sample, choose a solvent for each of the 8 types of solvents proposed by Snyder, they are petroleum ether, ethanol, tetrahydrofuran, acetone, methylene chloride, ethyl acetate, benzene and water Carry out conventional ultrasonic extraction with them as solvents respectively; Merge the extracts of 8 kinds of solvents, remove the solvent according to conventional methods, and obtain the galanga extract. Following the principle of like dissolves, using a combination of solvents to extract samples can make the extract more representative.

[0101] 2) The span and channel width of TLC correspond strictly to the 384-well square microplate array (the array format is 24 x 16 arrays in width of 4.5mm squa...

Embodiment 2

[0104] The whole chromatographic thin layer recorded with the planar chromatographic fingerprint is divided, stripped and positioned according to the square-mouthed microwell plate array format and eluted into the corresponding microwell pool of the microwell plate to prepare a planar chromatographic component microarray sample.

[0105] 1) The grid interface plate is prepared by cobalt-chromium alloy 3D printing, and the grid interface plate with the same array format as the 384 square microplate and the inner space is a square prism, specifically: 4.5mm×4.5mm square Horizontal 24×longitudinal 16 array format, the inner space of the grid is a square prism, the vertical section of the grid frame is an isosceles trapezoid with a height of 2.5mm, the width of the lower bottom of the isosceles trapezoid is the same as that of the 384 square mouth microporous plate microporous pool The thickness is the same, specifically 0.70mm, and this surface is used for docking with the microwe...

Embodiment 3

[0114] Cell Viability Assay on TLC Component Microarray Samples

[0115]On the galangal two-dimensional thin-layer chromatography component microarray sample prepared by the method of Example 2, the test solution was divided into the corresponding reaction pools of the 384-well square-mouth microplate (Corning 3701) according to the array coordinates to carry out Alamar Blue corporate lung cancer respectively. The survival rate of A549 cells and HepG2 liver cancer cells was detected, and 5-Fu (pentafluorouracil) was used as a positive control to detect the inhibitory effect of galangal TLC components on A549 lung cancer cells and HepG2 liver cancer cells. Referring to the instructions of the Invitrogen Alamar blue kit and conventional experimental methods, add 30 μL of cell suspension (logarithmic growth phase cells, 1000 cells / well) to each well, and place in a 37°C carbon dioxide cell incubator; Blue was added 12 hours before the detection; the measurement wavelength was 570...

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Abstract

The invention belongs to the field of active component screening, and relates to a method for high-throughput screening of bioactive components in complex components by "planar chromatographic component microarray". In the present invention, the planar chromatogram that reflects the molecular diversity and differential characteristics of complex components as a whole is prepared into a "planar chromatographic component microarray" sample, and the sample is used to carry out biological activity tests and focus on significant biologically active array units for mass spectrometry analysis; to have significant The dependence relationship between the biological activity intensity on the area of ​​the biological activity array unit and the biomass spectrum quasi-molecular ion peak intensity of the corresponding array unit and their coupling degree of chromatographic behavior are used as indicators to analyze the biological active ingredient. The invention does not need to separate and purify chemical components one by one in advance, does not need comprehensive analysis and detection, does not rely on molecular diversity compound library, samples can be amplified in parallel, good compatibility, high throughput, digitalization, greatly improves screening efficiency and reduces workload and cost.

Description

technical field [0001] The invention belongs to the technical field of drug analysis and biological testing, and specifically relates to a method for screening bioactive components in complex components with a high-throughput planar chromatographic component microarray and its application. Background technique [0002] Natural products, traditional Chinese medicines and their prescriptions, as well as synthetic and bioengineering products, such as various material components in terrestrial animals, plants, marine organisms, and microorganisms, as well as isolated biological secondary metabolites, are potential medicinal resource compound libraries. Most of them have the characteristics of complex components and large differences in content. Screening new drugs with significant biological activity has always been a challenge for the industry. [0003] High-throughput drug screening technology is developing rapidly as a means to quickly discover pharmaceutical lead compounds, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/90G01N30/88
CPCG01N30/90G01N30/88G01N2030/8813
Inventor 邓亦峰林振鹏
Owner GUANGDONG MEDICAL UNIV
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