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A sample pretreatment method for quantitative metabolomics analysis of adherent cells

A technology for sample pretreatment and metabolomics, which is applied in the field of sample pretreatment for quantitative metabolomics analysis of adherent cells, which can solve the problem of uneven buffer and achieve the effect of facilitating subsequent quantitative or semi-quantitative analysis.

Active Publication Date: 2017-06-06
上海裕隆生物科技有限公司
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  • Application Information

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Problems solved by technology

This method is mainly aimed at the problem of inhomogeneity of residual buffer in each culture dish. After washing the cells with buffer, the method of centrifuging with a 96-well plate centrifuge to remove the residual liquid can make almost no liquid residue in each culture dish. , for subsequent quantitative or semi-quantitative analysis

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  • A sample pretreatment method for quantitative metabolomics analysis of adherent cells
  • A sample pretreatment method for quantitative metabolomics analysis of adherent cells

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Embodiment １

[0027] The method of the invention

[0028]Using HCT-116, the LOVO cell line was cultured in DMEM high-glucose medium (purchased from HYCLONE) containing 10% FBS (purchased from Hangzhou Sijiqing and Harbin Saituo Company) at 37°C in an environment of 5% carbon dioxide. Discard the medium after 24 hours, then wash the cells with 4°C isotonic buffer salt for 3 times, add an appropriate amount of buffer salt to the culture dish for the fourth time, pour out the liquid manually, and place the culture dish upside down on a 96-well plate Centrifuge at 3500rpm on the rotor of the centrifuge for 1 minute. At this time, almost all the liquid in the petri dish is completely removed; then take 1 ml of methanol aqueous solution with a volume ratio of 1:1 containing a quantitative internal standard in the petri dish, and the internal standard of the methanol aqueous solution used is 50ug / ml oxalic acid, scrape the cells with a cell spatula, collect all the cell suspension into a 15 ml ce...

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Abstract

The invention discloses a method for pretreating adherent cell quantitative metabonomics analysis samples. The method mainly solves the problem that residual buffer solutions in all petri dishes are non-uniform, cells are washed with buffer solutions, and then the residual buffer solutions are removed by a 96-pore plate centrifuge in a centrifugal manner; with the adoption of the method, almost no solution residue exists in each petri dish, and follow-up quantitative or semi-quantitative analysis is facilitated.

Description

technical field [0001] The invention relates to the technical field of analytical chemistry, in particular to a sample pretreatment method for quantitative metabolomics analysis of adherent cells. Background technique [0002] Metabolomics is another research technology characterized by high throughput after genomics, transcriptomics and proteomics. It is mainly through the qualitative and quantitative analysis of small molecule metabolites in cells, tissues or specific body fluids. Analyze phenotypic characteristics of reactive biological systems. Metabolite molecules are the ultimate embodiment of gene and protein activities, and can best reflect the phenotypic characteristics of the research object. At present, they have been widely used in various fields of life sciences such as diseases, nutrition, metabolism, genetically modified crops, and microorganisms. Compared with other omics technologies, the advantages of metabolomics are at least reflected in the following as...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N30/06
Inventor 李盛高鹏刘赞赞华夏一凤
Owner 上海裕隆生物科技有限公司