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A high-throughput mass spectrometry method for the detection of primary metabolites in maize pollen

A technology for corn pollen and metabolites, applied in the field of chemical detection, can solve problems such as no reports, and achieve the effect of high stability and good repeatability

Active Publication Date: 2021-01-08
AGRO BIOLOGICAL GENE RES CENT GUANGDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the research on corn pollen is mostly focused on the determination of vigor, wall breaking, single or several chemical components analysis, health effects, etc., while the qualitative and quantitative analysis of primary metabolites in pollen (primary metabolome ) method research has not been reported yet

Method used

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  • A high-throughput mass spectrometry method for the detection of primary metabolites in maize pollen
  • A high-throughput mass spectrometry method for the detection of primary metabolites in maize pollen
  • A high-throughput mass spectrometry method for the detection of primary metabolites in maize pollen

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Embodiment 1

[0023]The present invention provides a technical solution: a mass spectrometry method for high-throughput detection of primary metabolites in corn pollen, including the following steps:

[0024]S1. Pretreatment of pollen samples: store the collected corn pollen in a liquid nitrogen tank at -196℃ for 30 minutes to quench the metabolic reaction, and then transfer it to the -80℃ refrigerator for storage for later use, and take it out of the refrigerator The pollen sample is placed in a mortar, frozen by adding liquid nitrogen and ground into a powder to break the wall, put into a 15mL centrifuge tube and stored in a refrigerator at -80℃ for analysis;

[0025]S2. Extraction of pollen metabolites: accurately weigh 50mg of corn pollen powder sample in a 2mL centrifuge tube, add 1mL of pre-cooled extract A and 10μL13C-labeled D-ribitol (1mg / mL) internal standard, mix well until the sample is completely resuspended, then place the sample on a shaker, shake for 10 minutes at 4℃, 300rpm, and then T...

Embodiment 2

[0030]The present invention provides a technical solution: a mass spectrometry method for high-throughput detection of primary metabolites in corn pollen, including the following steps:

[0031]S1. Pretreatment of pollen samples: store the collected corn pollen in a liquid nitrogen tank at -196℃ for 30 minutes to quench the metabolic reaction, and then transfer it to the -80℃ refrigerator for storage for later use, and take it out of the refrigerator The pollen sample is placed in a mortar, frozen by adding liquid nitrogen and ground into a powder to break the wall, put into a 15mL centrifuge tube and stored in a -80℃ refrigerator for analysis;

[0032]S2. Extraction of pollen metabolites: accurately weigh 50mg of corn pollen powder sample into a 2mL centrifuge tube, add 1mL of pre-cooled extract A and 10μL13C-labeled D-ribitol (1mg / mL) internal standard, mix well until the sample is completely resuspended, then place the sample on a shaker, shake for 10 minutes at 4℃, 300rpm, and then Th...

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Abstract

The invention discloses a mass spectrometry method for high-throughput detection of primary metabolites in corn pollen. A common corn pollen metabolome extracting solution is firstly quickly analyzedby using a gas chromatography-mass spectrometry technology to obtain a comprehensive metabolic profile spectrum. 67 primary metabolites contained in the pollen are systematically identified for the first time. Then, a target metabolome analysis method is established for 67 primary metabolites, and quantitative analysis can be conducted on the corn pollen primary metabolome at a time. The method comprises the steps of sample pretreatment, chromatography-mass spectrometry separation, metabolite result identification and metabolite quantitative analysis. The method has the advantages of good repeatability, high stability, rapidness and reliability, and is suitable for primary metabolome analysis of corn pollen and other crop pollen samples.

Description

Technical field[0001]The invention relates to the technical field of chemical detection, in particular to a mass spectrometry method for high-throughput detection of primary metabolites in corn pollen.Background technique[0002]Metabolism is a life process shared by organisms. Plant metabolism can be divided into two processes: primary metabolism and secondary metabolism. Primary metabolites are an important metabolite of organisms, which provide energy and intermediate products for the growth, development and reproduction of living organisms, and are also called nutrients. Metabolomics is the science of qualitatively quantifying all endogenous metabolites of cells in a biological system under given conditions, and then detecting the whole body and the law of response to changes in internal and external factors. Mainly based on GC-MS, LC-MS, NMR and other common technologies.[0003]Pollen is not only the fine particles in the stamens of plants, but also the male cells of hereditary pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 晏石娟黄文洁陈庄孔谦张群洁
Owner AGRO BIOLOGICAL GENE RES CENT GUANGDONG ACADEMY OF AGRI SCI
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