Targeting qualitative and quantitative metabonomic analysis method used for screening cancer biomarkers based on LC-MS/MS technology

A biomarker, quantitative analysis technology, applied in the field of metabolomics biomarker screening and discovery, can solve the limitations of data comparison, integration and definition of clinical indicators, constraints on the accuracy and reliability of biomarkers, unknown metabolism Problems such as difficulty in identifying the structure of

Inactive Publication Date: 2019-05-07
INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

At present, the discovery of small molecule biomarkers mainly adopts the method of non-targeted metabolomics. However, due to the limitation of research purpose and technical means, non-targeted metabolomics also has some shortcomings, such as: only using peak area or response The lack of accurate quantification of changes in metabolites characterized by intensity, which limits the comparison and integration of data between laboratories and the definition of clinical indicators; the collection method of mass spectrometry full scan leads to the loss of information on some low-level metabolites; the standard and standard database Lack leads to difficulty in identification of unknown metabolite structure, etc. These problems restrict the accuracy and reliability of finding possible biomarkers

Method used

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  • Targeting qualitative and quantitative metabonomic analysis method used for screening cancer biomarkers based on LC-MS/MS technology
  • Targeting qualitative and quantitative metabonomic analysis method used for screening cancer biomarkers based on LC-MS/MS technology
  • Targeting qualitative and quantitative metabonomic analysis method used for screening cancer biomarkers based on LC-MS/MS technology

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Embodiment 1

[0021] 1. Establish a liquid chromatography-mass spectrometry quantitative analysis method for 106 core metabolites in plasma. The specific sample pretreatment conditions and measurement conditions are:

[0022] (1) Sample pretreatment: Frozen plasma samples were thawed at 4°C and vortexed on ice to mix well. Accurately draw 100 μL of plasma, add 20 μL of standard mixed solution, mix 10 μL of internal standard, and vortex to mix. Add 300 μL of refrigerated acetonitrile, vortex at 3000rpm for 240s, centrifuge at 10000r / min at 4°C for 10min, separate the supernatant, concentrate it by centrifugation for 2 hours, add 100μL of acetonitrile:water (3:7, V / V) solution to redissolve , vortex at 3000rpm for 240s, centrifuge at 10000r / min for 10min, separate the supernatant, filter through a 96-well plate and inject the sample.

[0023] (2) The mixed internal standard solution is prepared according to the following method: Accurately weigh 1.0 mg of the isotopic internal standard produ...

Embodiment 2

[0064] Through the previous plasma metabolomics research on animal models of esophageal cancer in our laboratory, 61 small molecule metabolites related to rat esophageal cancer were found, which were called core metabolites in this study. In order to extend the research results to the detection of clinical samples and overcome the lack of metabolite information caused by the species differences between rats and humans, the technical solution of the present invention was adopted to establish a target for clinical plasma biomarker screening. Qualitative and quantitative metabolomics analysis methods.

[0065] 1. Establish a liquid chromatography-mass spectrometry semi-quantitative analysis method for core metabolites in clinical plasma. The specific sample pretreatment conditions and measurement conditions are:

[0066] (1) Sample pretreatment: Frozen plasma samples were thawed at 4°C. and vortexed on ice to mix. Accurately draw 100 μL of plasma, refrigerate 300 μL of acetonit...

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Abstract

The invention relates to a targeting qualitative and quantitative metabonomic technical analysis method used for screening cancer biomarkers based on the LC-MS/MS technology. The method is characterized in that a structure prediction function of multi-reaction monitoring-dependent information acquisition-enhanced daughter ion detection on correlative metabolites and a structure identification function of a high-resolution mass spectrum are predicted by fully utilizing a multi-reaction monitoring quantitation function of the high performance liquid-tandem mass spectrum, and the analysis methodsuitable for screening biomarkers of various cancers is formed. The method has the advantages of high flux, good stability and high adaptability, and improves sensitivity and accuracy of screening andfinding of metabonomic biomarkers.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a method for screening and discovering metabolomics biomarkers based on liquid chromatography-mass spectrometry analysis technology. Background technique [0002] In recent years, the study of malignant tumors based on metabolomics has attracted much attention, and it has been found that many cancers are closely related to the early diagnosis and efficacy evaluation of malignant diseases such as lung cancer, esophageal cancer, prostate cancer, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, and liver cancer. Associated possible small molecule biomarkers. At present, the discovery of small molecule biomarkers mainly adopts the method of non-targeted metabolomics. However, due to the limitation of research purpose and technical means, non-targeted metabolomics also has some shortcomings, such as: only using peak area or response The lack of accurate quantific...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72G01N30/86
Inventor 再帕尔阿不力孜陈艳华岳小飞周帜徐婧张瑞萍
Owner INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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