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A Quasi-targeted Metabolomics Analysis Method Based on Liquid Chromatography/Mass Spectrometry

A technology of metabolomics and analytical methods, applied in analytical materials, scientific instruments, material separation, etc., to achieve the effect of wide detection linear range, good detection repeatability and response linearity, and improved accuracy

Active Publication Date: 2017-02-08
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • 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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  • A Quasi-targeted Metabolomics Analysis Method Based on Liquid Chromatography/Mass Spectrometry
  • A Quasi-targeted Metabolomics Analysis Method Based on Liquid Chromatography/Mass Spectrometry
  • A Quasi-targeted Metabolomics Analysis Method Based on Liquid Chromatography/Mass Spectrometry

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 / QQQMS-DMRM platform. ...

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Abstract

The present invention is a pseudo-target metabolomics analysis method based on liquid chromatography / mass spectrometry. The samples to be analyzed are respectively prepared into combined samples according to the groups, and the ultra-high performance liquid chromatography / quadrupole-time-of-flight mass spectrometry is used for the analysis. The data-dependent acquisition mode automatically collects the secondary mass spectrometry of the metabolites contained in each combined sample, and uses qualitative analysis software to extract the retention time, parent ion and product ion information of the measured metabolites, and screen out the metabolites according to the response intensity of the product ions. The characteristic transition information is added, and the characteristic transition information obtained from each combined sample is summed. Then, in the actual sample, UHPLC / QQQ MS is used to scan the obtained metabolite characteristic ion pairs through dynamic multiple reaction monitoring mode to obtain the corresponding spectrum, and then integrated through quantitative analysis software to obtain the metabolite contained in the sample to be tested. information on the substance and its content.

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 Patents(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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