Simulative-target metabonomics analytic method based on combination of liquid chromatography and mass spectrum

A technology of metabolomics and analysis methods, applied in the direction of analysis of materials, scientific instruments, material separation, etc., to ensure reliability, simplify the process of data processing, and improve accuracy.

Active Publication Date: 2015-01-21
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Application Information

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Problems solved by technology

However, a big limitation of metabolite target analysis is that it can only analyze a small number of known metabolites, which is far from the goal of metabolomics analysis of as many metabolites as possible.

Method used

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  • Simulative-target metabonomics analytic method based on combination of liquid chromatography and mass spectrum
  • Simulative-target metabonomics analytic method based on combination of liquid chromatography and mass spectrum
  • Simulative-target metabonomics analytic method based on combination of liquid chromatography and mass spectrum

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1: Establishment of UHPLC / QQQ MS-DMRM-based Metabolomics Analysis Method for Serum Pseudo-targets

[0026] The process of establishing the metabolomics analysis method based on UHPLC / QQQ MS-DMRM for serum quasi-targets is shown in the attached figure 1 As shown, the specific implementation steps are as follows:

[0027] 1. Combined sample making and preprocessing

[0028] 50 μL of sera from 29 cases of liver cancer patients were pipetted in equal amounts and pooled, and at the same time, 50 μL of sera from 30 normal people were pipetted in equal amounts and combined. Pipette 200 μL of combined serum from liver cancer patients and normal subjects, add 800 μL of acetonitrile to separate proteins, and pipette 800 μL of supernatant to freeze-dry. The freeze-dried product was redissolved in 100 μL pure water for injection analysis.

[0029] 2. Automatic MS / MS data acquisition of metabolites

[0030]Ultra-high performance liquid chromatography (Agilent1200RRLC) / qu...

Embodiment 2

[0041] Example 2: Methodological investigation of metabolomics analysis of serum quasi-targets based on UHPLC / QQQ MS-DMRM

[0042] Mix equal volumes of the pooled serum of normal people and the pooled serum of patients with liver cancer to form a quality control (QC) sample, and perform pretreatment according to the following different objects.

[0043] 1. Repeatability inspection of analytical methods

[0044] Pipette 100 μL of QC serum, add 400 μL of acetonitrile to remove protein, and pipette 400 μL of supernatant to lyophilize. The freeze-dried product was redissolved in 100 μL pure water for injection analysis. Repeat processing of 10 QC serum samples was done in parallel.

[0045] The 10 QC samples were analyzed by metabolomics on UHPLC / Q-TOF MS and UHPLC / QQQ MS-DMRM platforms respectively. The data measured on the UHPLC / Q-TOF MS platform was matched by XCMS software, and 318 peaks were obtained that were identical to those measured on the UHPLC / QQQ MS-DMRM platform. ...

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Abstract

The invention provides a simulative-target metabonomics analytic method based on combination of liquid chromatography and mass spectrum. The method includes following steps: respectively manufacturing to-be-analyzed samples into merged samples according to groups; automatically collecting secondary mass spectrums of metabolites in each merged samples through data of UHPLS / Q-TOF MS with dependence on a collecting mode; extracting a retaining time and information of parent ions and daughter ions of the metabolites by qualitative analytic software; screening out characteristic ion pair information of the metabolites according to daughter ion response intensity; summing the characteristic ion pair information of each merged samples; and scanning the obtained characteristic ion pair of the metabolites by UHPLC / QQQ MS through a dynamic multi-reaction monitoring mode in an actual sample for obtaining a corresponding spectrogram; and performing integration through quantitative analytic software to obtain the metabolites in the to-be-analyzed samples and content information thereof.

Description

technical field [0001] The invention relates to the field of analytical chemistry, in particular to a method for metabolomics analysis of pseudo-targets based on ultra-high performance liquid chromatography / triple quadrupole mass spectrometry dynamic multiple reaction monitoring mode. Background technique [0002] Metabolomics is another important branch after genomics, transcriptomics and proteomics in the development of systems biology. To study the biological phenomena and their functions related to the metabolic phenotype. Metabolomics technology is widely used in disease-related research to discover metabolic markers related to disease diagnosis, study disease pathogenesis, and evaluate surgical effects and drug efficacy for disease treatment. Commonly used metabolomics platforms are mainly based on nuclear magnetic resonance technology, mass spectrometry and their combined techniques. NMR technology has the advantages of simple sample preparation and high throughput,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 许国旺陈世礼孔宏伟路鑫李勇
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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