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Cell lipidomics analysis method based on ultra-high performance liquid chromatography-tandem high-resolution mass spectrometry

An ultra-high performance liquid phase, high-resolution mass spectrometry technology, applied in analytical materials, scientific instruments, material separation, etc., can solve problems such as inability to compare lipid types and content, unclear understanding of lipid existence, etc., and achieve savings. Effects of time and economic costs, wide lipid coverage, optimal chemical stability

Inactive Publication Date: 2020-09-18
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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Problems solved by technology

[0004] In view of the important role of lipid compounds in cells, it is urgent to establish a method for the detection of lipids that can be widely used in different cells. The existing detection methods are only for individual types of cells, but for more types of cells. The presence of lipids is not well understood, and it is not possible to compare the type and amount of lipids in different types of cells

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  • Cell lipidomics analysis method based on ultra-high performance liquid chromatography-tandem high-resolution mass spectrometry
  • Cell lipidomics analysis method based on ultra-high performance liquid chromatography-tandem high-resolution mass spectrometry
  • Cell lipidomics analysis method based on ultra-high performance liquid chromatography-tandem high-resolution mass spectrometry

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Embodiment 1

[0057] 1. Sample preparation and processing: The normal growth of PC12 cells, MCF-7 cells, HepG2 cells, RKO-E6 cells, and Hela cells were adjusted to 5 × 10 cells with PBS. 5 Cells / mL, the cells were broken by ultrasonic method (conditions: ultrasonic for 3s, stop working for 5s, and repeat this cycle for 60 times). Take 100 μL of the sonicated cell suspension, add 300 μL of extraction reagent (methanol:dichloromethane=2:1, v / v), vortex for 30s, centrifuge at 5000rpm for 5min, take out the supernatant and transfer it to a glass tube, add The extraction was repeated once with 300 μL of extraction reagent, and the organic phases of the two extractions were combined. The combined organic phases were blown dry with nitrogen, then reconstituted with 0.5 mL of methanol, and passed through a Hippoch 0.2 μm PTFE filter.

[0058] 2. According to the analysis conditions of the ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap Fourier transform mass sp...

Embodiment 2

[0065] 1. Sample preparation and processing: The normal growth of PC12 cells, MCF-7 cells, HepG2 cells, RKO-E6 cells, and Hela cells were adjusted to 1×10 with PBS. 6 Cells / mL, the cells were broken by ultrasonic method (conditions: ultrasonic for 3s, stop working for 5s, and repeat this cycle for 60 times). Take 100 μL of the sonicated cell suspension, add 300 μL of extraction reagent (methanol:dichloromethane=2:1, v / v), vortex for 30s, centrifuge at 5000rpm for 5min, take out the supernatant and transfer it to a glass tube, add The extraction was repeated once with 300 μL of extraction reagent, and the organic phases of the two extractions were combined. The combined organic phases were blown dry with nitrogen, then reconstituted with 0.5 mL of methanol, and passed through a Hippoch 0.2 μm PTFE filter.

[0066] 2. According to the analysis conditions of the ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap Fourier transform mass spectromet...

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Abstract

The invention provides a cell lipidomics analysis method based on ultra-high performance liquid chromatography tandem high-resolution mass spectrometry, and relates to the field of analytical chemistry and the field of lipidomics research. The method comprises the following steps: adjusting the density of cells with PBS, and carrying out ultrasonic disruption to obtain a cell suspension; extracting the cell suspension with methanol and a dichloromethane solution to obtain a sample to be detected; separating and determining the sample to be detected through ultra-high performance liquid chromatography-quadrupole rod-electrostatic field orbitrap Fourier transform mass spectrometry; and carrying out qualitative analysis on the measured lipid compound by utilizing software. According to the present invention, the method has characteristics of simple operation and wide lipid coverage, does not require the standard substance, and can be effectively used for the lipidomics analysis of different types of cells so as to further develop the biological effect research on cytotoxicity or activity and the like.

Description

technical field [0001] The invention relates to the fields of analytical chemistry and lipidomics research, in particular to a cell lipidomics analysis method based on ultra-high performance liquid chromatography tandem high-resolution mass spectrometry. Background technique [0002] Cells are the basic units of the structure and function of living organisms, and they are extremely small in size and various in shape. Cells are mainly composed of nucleus and cytoplasm, with a cell membrane on the surface, and a cell wall outside the cell membrane of higher plants. The chemical substances that make up cells are divided into inorganic substances and organic substances. There are thousands of organic substances in cells, accounting for more than 90% of the dry weight of cells. There are four main types of molecules in organic substances, namely proteins, nucleic acids, lipids and carbohydrate. [0003] Lipids play an important role in cells, such as participating in energy met...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8813
Inventor 邱静王昕璐贾琪钱永忠
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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