Method for identifying lipid component structure in lipidomics based on CID fragmentation
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A technology of lipid composition and lipidome, applied in the field of life sciences, which can solve problems such as difficulties and bottlenecks
Inactive Publication Date: 2017-10-03
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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Problems solved by technology
Although the development of analytical technology has brought many benefits to the analysis of lipid compounds, there is a bottleneck in the further determination of lipid molecular structure in lipid analysis based on mass spectrometry
Mass spectrometry brings a large amount of spectrogram data, analysts need to spend a lot of time and energy to analyze these spectrograms, and this work is also difficult for most life scientists
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Embodiment 1
[0021] Composition Determination of Triglycerides in Human Plasma Samples
[0022] a: Extraction of lipid components in plasma samples
[0023] Select the Folch extraction method to extract triglycerides in the plasma sample, as follows: add 180 μL methanol to 20 μL plasma sample and vortex fully, then add 400 μL chloroform, vortex the sample, and finally introduce 150 μL water layer. After sufficient extraction, centrifuge at 8000 g for 10 min at 4°C. Remove the lipid-rich chloroform layer. The chloroform solvent was evaporated to dryness. Reconstitute with isopropanol / methanol solvent. Centrifuge and take the supernatant for injection.
[0024] b: Select the appropriate detection method and detection conditions
[0025] Chromatographic separation conditions: Chromatographic column: Hypersil GOLD aQ column (2.1×150mm, 3μm I.D.). Mobile phase: A is acetonitrile / water solution containing 0.1% formic acid and 5mM ammonium formate (40:60, V / V), B is isopropanol / acetonitrile...
Embodiment 2
[0030] Determination of phosphatidylcholine in lung tissue samples of SD rats
[0031] a: Extraction of lipid components in liver tissue samples
[0032] Select the methyl tert-butyl ether-methanol extraction method (MTBE extraction method) to extract the components of phosphatidylcholine in the liver tissue sample, as follows: After the 50 mg liver sample is homogenized, add 400 μL of pre-cooled 75% methanol solution, vortex After vortexing for 10 minutes, add 1 mL of methyl tert-butyl ether, vortex for 20 minutes, then add 60 μL Milli-Q ultrapure water, vortex for 10 minutes, let stand for 10 minutes, centrifuge at 17,000 rpm for 10 minutes, separate the samples, and take 900 μL of the supernatant and centrifuge The concentrator was evaporated to dryness. The upper layer concentrate was redissolved in 100 μL (isopropanol:methanol=8:2) mixture, sonicated in a water bath for 10 minutes, centrifuged at 17,000 rpm for 10 minutes, and 40 μL of the supernatant was taken for testi...
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Abstract
The present invention discloses a simple method for identifying unknown lipid components in lipidomics based on CID fragmentation. The main technical route comprises: 1, establishing a lipidomics analysis and determination method; 2, establishing the lipidomics analysis and determination method by using a multi-stage fragmentation mass spectrometry hybrid high resolution mass spectrometer; 3, searching an online spectrum library according to the accurate mass number, and attributing the lipid component types corresponding to the lipid chromatographic peaks to be identified; 4, determining the total carbon content and the unsaturation degree of the fatty acyl part in the lipid molecule; 5, according to a simple formula, determining the characteristic fragment information of each chromatographic peak; 6, deriving the carbon number and the unsaturation degree of each fatty acyl group according to the obtained characteristic fragment information; and 7, finally determining the lipid molecule structure. According to the present invention, the method is accurate and sensitive, is independent of the standard compound, and can rapidly identify the structure of the unknown lipid molecule in the lipidomics data.
Description
Technical field: [0001] The invention relates to life sciences, in particular to the establishment of metabolomics and lipidomics analysis and determination methods and the rapid identification of unknown lipid component structures. Background technique [0002] Due to the integrity and complexity of living organisms, the development of modern biology has returned to a new level of integrative research from organs, tissues, cells to genes, which means that the era of molecular biology has entered the era of systems biology. Therefore, systems biology research methods such as genomics, transcriptomics, proteomics and metabolomics have become new means to interpret the mysteries of life sciences and clarify the mechanism of physiological and pathological changes and drug intervention. [0003] Lipidomics is a system biological analysis method that has developed rapidly in recent years. It is a new omics method for studying biological systems developed after metabolomics and pr...
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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N27/62
CPCG01N30/02G01N27/626
Inventor 谢彤单进军徐建亚沈存思狄留庆汪受传
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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