Large-scale plasma lipid targeted quantitative analysis method based on liquid chromatography-mass spectrometry

A liquid chromatography and quantitative analysis technology, which is applied in the field of large-scale quantitative analysis of plasma lipids, can solve problems such as uncorrectable, inaccurate quantitative results, and inability to solve the problems of the accuracy of quantitative results of nonlinear signal changes, and achieve low solvent toxicity. , Simple and fast operation

Pending Publication Date: 2021-11-09
武汉迈特维尔生物科技有限公司
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Problems solved by technology

For relative quantification, the quantitative results are inaccurate due to the inability to correct the deviation of pre-processing and mass spectrometry analysis in the experimental process.
Similarly, the absolute quantitativ

Method used

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  • Large-scale plasma lipid targeted quantitative analysis method based on liquid chromatography-mass spectrometry
  • Large-scale plasma lipid targeted quantitative analysis method based on liquid chromatography-mass spectrometry
  • Large-scale plasma lipid targeted quantitative analysis method based on liquid chromatography-mass spectrometry

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Example Embodiment

[0037] Example 1

[0038] A targeted quantitative analysis method for human plasma lipids, comprising the following steps:

[0039] S1, sample processing:

[0040] Step 1: After thawing the cryopreserved sample, vortex and mix well, pipette 20 μL of the sample and add it to a 1.5 mL centrifuge tube with the corresponding number.

[0041]Step 2: Add 20 μL internal standard mixed solution, and vortex to mix. The internal standard mixture contains 24 isotope internal standards (FA(16:0)-d31 10μmol / L, Palmitoyl-L-carnitine-(N-methyl-d3)2μmol / L, 5S-HETE-d810μmol / L, cholesterol -d7 250μmol / L, 18:1Chol(d7)ester 100μmol / L, GCDCA-d4 10μmol / L, sphingosine-d7 2μmol / L, Ceramide(d18:1(d7) / 18:0)5μmol / L, Ceramide- 1-Phosphate(d18:1(d7) / 15:0) 10μmol / L, C18 Glucosyl(β)Ceramide-d5 20μmol / L, SM(d18:1(d9) / 15:0) 10μmol / L, DG( 17:0 / 17:0)-d5 20μmol / L, TG(17:0 / 17:1 / 17:0)-d5 10μmol / L, LPC(16:0)-d3110μmol / L, LPE(18:1 )-d5 10μmol / L, LPG(15:0)-d5 10μmol / L, LPG(15:0)-d5 10μmol / L, LPI(17:0)-d5 10μmol / ...

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Abstract

According to the method, the external standard curve corrected by the internal standard is adopted for quantitative analysis of the concentration of the lipid, so that the defects that the deviation of pretreatment and mass spectrometry in the experiment process cannot be corrected and the actual lipid concentration change relationship cannot be accurately reflected in a pure peak area relative quantitative method are overcome; meanwhile, the problem that the nonlinear signal change of actual lipid on a mass spectrometer cannot be solved in the quantification process of an internal standard method of a single internal standard or multiple internal standards is also solved; the methyl tert-butyl ether-methanol extracting solution is adopted for extraction, pretreatment is carried out in the mode that isopropanol and acetonitrile are redissolved according to the ratio of 1: 1, operation is easy and rapid, solvent toxicity is small, and the method can be suitable for large-scale sample analysis.

Description

technical field [0001] The invention relates to analytical chemistry and biological detection, and is a large-scale plasma lipid quantitative analysis method based on liquid chromatography-mass spectrometry. Background technique [0002] Lipid is a class of insoluble small molecular substances, mainly divided into fatty acids, glycerolipids, phospholipids or cholesterol. Lipids are widely distributed in human cells and blood. Lipids are components of cell membranes and play an important role in energy storage and signaling molecules in the body. In human blood, lipid accounts for more than 65% of all substances and is an important component of blood. Due to the convenience of blood collection, it is the most common sample source in lipid research. At present, the lipid analysis of human blood is a research hotspot in the screening of cancer research markers, the body's metabolic mechanism and clinical diagnosis. [0003] The current lipid analysis mainly relies on liquid c...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 方聪明陈宇唐堂
Owner 武汉迈特维尔生物科技有限公司
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