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A Relative Quantitative Analysis Method for Metabolomics Based on UPLC/HRMS

A metabolomics and relative quantitative technology, applied in the field of relative quantitative detection of small molecule metabolites, can solve the problems of unclear metabolite concentration changes, incompatibility, and little significance of metabolomics, and achieve simple and easy pretreatment, Simple operation and wide applicability

Active Publication Date: 2021-11-05
SUZHOU BIONOVOGENE BIOMEDICAL TECH CO LTD
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Problems solved by technology

Although the above two detection methods have solved the qualitative and quantitative problems to a certain extent, they still need to use two mass spectrometry platforms at the same time, and both use the quality control serum (QC) samples of their respective laboratories as the quantitative calibration samples for calculating the concentration. As a result, the concentration results measured by each laboratory are independent and incompatible
It is especially important that the above-mentioned method does not identify (or annotate) the structure of metabolites or differential metabolites. Fundamentally solve the simultaneous qualitative and quantitative technical problems of metabolomics

Method used

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  • A Relative Quantitative Analysis Method for Metabolomics Based on UPLC/HRMS
  • A Relative Quantitative Analysis Method for Metabolomics Based on UPLC/HRMS
  • A Relative Quantitative Analysis Method for Metabolomics Based on UPLC/HRMS

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Embodiment

[0084] Example: Based UPLC / HRMS healthy subjects and patients with cerebral infarction serum Metabolomics A method for analyzing the relative quantitative.

[0085] Establish process-based UPLC / HRMS healthy subjects and patients with cerebral infarction serum metabolomics semi-quantitative analysis methods such as figure 1 As shown in the specific embodiment the following steps:

[0086] 1, internal standard was prepared:

[0087] 1.1 Preparation of the mixed solution isotope internal standard:

[0088] Appropriate amount of D- fructose -1-13C (F-13C), L- valine -1-13C (Val-13C), choline chloride - trimethyl -d9 (Choline-d9), 5'- adenosine triphosphate -15N5 (ATP-15N5), L- phenylalanine -1-13C (Phe-13C), uridine -2-13C (U-2-13C), vitamin B3-d4 (VB3-d4), L carnitine hydrochloride - methyl -d3 (L-carnitine-d3) and betaine hydrochloride - trimethyl -d9 (betaine-d9) of 9 isotope internal standards were formulated as water or methanol was added 1 15mg / mL internal standard monoiso...

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Abstract

The invention belongs to the technical field of metabonomics, and relates to a relative quantitative analysis method of metabolomics based on UPLC / HRMS. Specifically, the method mainly includes the preparation of isotope internal standard mixed solution, standard curve calibration solution and metabolomics sample solution, obtaining the original mass spectrometry data of the standard curve calibration solution and metabolomics sample solution, and transposing and solving the original data. Convolve, identify and select the optimal isotope internal standard, fit the linear equation and calculate the relative quantitative results of metabolites in the metabolomics sample solution, and complete the structural identification of metabolites and differential metabolites in the metabolomics sample solution. The method can meet both qualitative and quantitative requirements with only a high-resolution mass spectrometer platform, with accurate quantitative results, high accuracy of qualitative results, low cost, simple operation, and wide applicability.

Description

technical field [0001] The invention belongs to the technical field of metabolomics, and relates to a method for relative quantitative detection of small molecule metabolites based on UPLC / HRMS and utilizing isotope internal standards. Background technique [0002] Metabolomics is the study of all metabolic responses in biological systems that are affected by factors such as exogenous substance stimulation, environmental changes, or genetic modifications, and detects the overall picture and dynamic changes of metabolic responses. 1000 small molecule metabolites, characterized by high-throughput, large-scale experimental methods combined with computational systems such as pattern recognition and expert systems, qualitative and quantitative analysis of all metabolites in organisms is an important part of systems biology. [0003] According to different research purposes, metabolomics can be divided into four levels: metabolite target analysis, metabolic profiling analysis, met...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/86
CPCG01N30/02G01N30/8696G01N30/8651G01N30/8655G01N30/8679G01N2030/045G01N2030/8831G01N30/72G01N30/8686G01N30/34G01N30/06
Inventor 徐英任建洪谢桂纲戴贞丽顾梦南谷从顺
Owner SUZHOU BIONOVOGENE BIOMEDICAL TECH CO LTD
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