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Targeted metabo lomics analysis method for determining metabolites of living body

A technology of metabolites and metabolomics, applied in the direction of analyzing materials, material analysis by electromagnetic means, measuring devices, etc., can solve problems such as gaps, and achieve the effect of overcoming complex processing

Inactive Publication Date: 2016-02-03
TIANJIN SUNNYPEAK BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional LC-MS / MS-based targeted metabolomics method has a big limitation that it can only measure a small number of metabolites (<50) at a time, which is different from the metabolomics analysis of as many metabolites as possible. There is a big gap in the target
In addition, the traditional metabolomics chromatography analysis time is more than 60 minutes

Method used

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  • Targeted metabo lomics analysis method for determining metabolites of living body
  • Targeted metabo lomics analysis method for determining metabolites of living body
  • Targeted metabo lomics analysis method for determining metabolites of living body

Examples

Experimental program
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Effect test

Embodiment 1

[0029] The following example 1 takes corn root as an example, but the method of the present invention is also applicable to solid samples such as food, animal tissue, and soil samples.

[0030] Establishment of a new-generation targeted metabolomics analysis method based on LC-MS / MS for the simultaneous determination of 700 metabolites in maize roots

specific Embodiment approach

[0031] The flow chart of the establishment and application of the new generation targeted metabolomics method is shown in figure 1 , the specific implementation is as follows:

[0032](1) Optimization of metabolite characteristic parent ions and fragment product ions: 700 standard compounds of compounds were dissolved in methanol or water according to their solubility to make a 1 μM solution. Take 1ml of the solution and inject it into the triple quadrupole mass spectrometer (ABSCIEX, Qtrap5500) through a syringe; turn on the mass spectrometry workstation, pass through Q1, MS 2 and MRM scanning to determine the best precursor ion of metabolites, fragment product ions, dissociation voltage, bombardment voltage and collision port exit voltage, and select the ion with the strongest scanning signal in Q1 as the best precursor ion for detection, MS 2 The ion with the strongest scanning signal is the best fragment product ion, and the best precursor ion and product ion are taken as...

Embodiment 2

[0055] Investigation of the accuracy of the method for the determination of metabolites in serum using a new generation of targeted metabolomics analysis method (taking serum as an example, applicable to liquid samples such as microbial liquid, urine, sewage, etc.)

[0056] (1) Metabolite extraction: take 95 μl of human serum sample, add 5 μl of Escherichia coli-yeast 13 C Stable isotope internal standard compound, after mixing, add 400 μl of extraction solvent (methanol-acetonitrile-water 40:40:20, v / v / v), vortex and mix well, and let stand on ice for 10 minutes. After high-speed low-temperature centrifugation (16000g, 4°C), the supernatant was taken to obtain the sample solution for LC-MS / MS analysis. Take the same serum and do 10 repeated samples in parallel.

[0057] (2) Investigation on method reproducibility: 10 μl of serum extract was taken, and 700 metabolites in 10 serum samples were determined by using the analytical method established above. Select 70 representati...

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Abstract

The invention provides a targeted metabo lomics analysis method for determining metabolites of a living body. The targeted metabo lomics analysis method can simultaneously determine 700 main metabolites of a living body based on series connection of ultra-high performance liquid chromatography and triple quadrupole and comprises the following steps: firstly, 700 main metabolites in a living body are screened; standard products of compounds are optimized to obtain optimal parent ions, daughter ions and mass spectrography conditions of the metabolites; the metabolites in samples are extracted by methanol, acetonitrile and water; liquid chromatograph conditions are optimized; a hydrophilic interaction is adopted to separate the metabolites; the separated metabolites enters a mass spectrum; the 700 main metabolites are determined at the same time by use of a real-time scheduled multi-reaction monitoring mode with quick positive and negative polarity switching function; the metabolite differences between all the samples are analyzed via partial least square analysis; changes of the metabolic pathway are found by pathway enrichment analysis. The targeted metabo lomics analysis method remarkably increases the number of detected compounds, and greatly shortens the analysis time.

Description

technical field [0001] The invention relates to the field of analytical chemistry, in particular to a targeted metabolomics analysis method for measuring metabolites of organisms. Background technique [0002] Metabolites are small molecular organic compounds (molecular weight <1500Da) that realize the metabolic process in the living body, and are the final products of gene expression generated under the catalysis of enzyme proteins. Metabolomics technology (metabolomics) is an important research method of systems biology developed in the post-gene era, which aims to quantitatively detect as many metabolites as possible in biological systems. Metabolomics technology is widely used in many fields, such as disease diagnosis, drug development, nutritional food evaluation, toxicology research, environmental monitoring, plant breeding, etc. [0003] Metabolomics technology is divided into non-targeted metabolomics and targeted metabolomics according to whether the detected me...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N27/62
Inventor 李克峰王旭
Owner TIANJIN SUNNYPEAK BIOTECH CO LTD
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