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Sample extraction method for research on tissue sample metabolomics and lipidomics

A technology of tissue samples and metabolomics, applied in the field of analytical chemistry, can solve the problem of small consumption of samples, achieve the effect of reducing the degradation loss of metabolites, ensuring stability and high extraction efficiency

Active Publication Date: 2018-09-28
SHANGHAI MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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Problems solved by technology

[0005] The purpose of the present invention is to address the deficiencies in the prior art, to provide a new method for two-step continuous extraction of hydrophilic and lipophilic metabolites in trace tissue samples, compared to the traditional method of using two samples to complete the extraction of hydrophilic and lipophilic metabolites The method has the advantages of small sample consumption and high throughput, which not only saves manpower, material and financial resources, but also saves time. Compared with similar methods reported in the literature, this method has higher extraction efficiency and good stability. Hydrophilic and lipophilic metabolites are in the Better distribution is achieved in the two extraction systems, avoiding the chromatographic separation problems caused by the partial distribution of partially hydrophilic metabolites to the lipid extraction layer during the phase separation process

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  • Sample extraction method for research on tissue sample metabolomics and lipidomics
  • Sample extraction method for research on tissue sample metabolomics and lipidomics
  • Sample extraction method for research on tissue sample metabolomics and lipidomics

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1 Rat Dried Sample Hydrophilic and Lipophilic Sample Extraction and Analysis

[0031] A) A normal rat liver was used, and 18 samples were weighed in a stoppered centrifuge tube, each sample 200-300mg.

[0032] B) According to the solvent ratio of 100 μL per 28 mg tissue sample, add an appropriate volume of hydrophilic metabolite extraction solution frozen MeOH-water (1 / 1, v / v), and then add MeOH and MTBE respectively to the centrifuge tube The cleaned steel balls were sealed and placed on dry ice for 30 s, and immediately placed in a fully automatic sample rapid grinder to grind twice, with a grinding frequency of 60 Hz and a grinding time of 20 s.

[0033] C) After grinding, immediately take out the sample and centrifuge at 13,000rpm at low temperature (4°C) for 20min. Concentrated in a centrifugal rotary evaporator (operating temperature: 45° C., vacuum: 0.1 psi, rotation speed: 3000 rpm) to dryness, as the hydrophilic fraction extract.

[0034] D) After tak...

Embodiment 2

[0040] Embodiment 2 different extraction method effect contrasts

[0041] There have been a large number of research reports on the extraction of the hydrophilic part of MeOH-H2O, and the data of the present invention are basically consistent with these reports. The present invention is mainly aimed at continuing to extract the lipid portion of the remaining tissue residue after the hydrophilic portion is extracted, and the relevant verification data is mainly in the aspect of lipid extraction. In order to highlight the advantages of the present invention, after the hydrophilic portion is extracted, MTBE (the present invention method) and literature report method-DCM-MeOH (3:1, v / v) (abbreviation DCM-MeOH method) carry out synchronous analysis, to evaluate the superiority of the present invention (see the attached document for details Figure 1-3 ).

[0042] (1) Comparison of data profile analysis of two lipidomics extraction methods in rat liver, such as figure 1 shown. Th...

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Abstract

The invention discloses a sample extraction method for research on tissue sample metabolomics and lipidomics. The sample extraction method for research on tissue sample metabolomics and lipidomics comprises the following steps: adding a trace amount of tissue sample into a pre-frozen methanol-aqueous solution in advance, performing low-temperature bead collision and homogenization, centrifuging totake supernatant for research on metabolomics, adding pre-frozen tert-butyl methyl ether into tissue residues, and homogenizing again and centrifuging for research on lipidomics. Hydrophilic and lipophilic endogenous metabolites can be extracted simultaneously through a trace amount of tissues and are applied to research on metabolomics and lipidomics correspondingly. Compared with the method reported by literatures, the method has the following advantages: extraction is conducted separately through two steps, so that the technical problems that lipid full spectrum coverage is difficult to realize by a single extraction system in the processes of extracting by a mixed solvent through one step and then extracting the hydrophilic and lipophilic metabolites through two-phase separation as well as part of the hydrophilic metabolites is distributed to a lipid extraction layer in the phase separation process are avoided; meanwhile MTBE extraction has higher lipid extraction efficiency and more convenient experimental operability than those of dichloromethane-methanol.

Description

technical field [0001] The invention relates to the technical field of analytical chemistry, in particular to a sample extraction method for tissue sample metabolomics and lipidomics research. Background technique [0002] Tissue samples, especially trace case samples and small animal tissue samples, when performing biochemical indicators, immunity, tissue sections, genes and proteins, and omics analysis, often encounter that the amount of samples is not enough for so many test and analysis. It is necessary to establish some high-throughput analysis methods suitable for the same sample that can simultaneously obtain multiple detection parameters. Metabolomics and lipidomics are methods for systems biology research. These two omics can obtain interrelated metabolic indicators. The same sample is used for two-group research at the same time, which can avoid taking two samples for two Due to the differences in sampling positions brought about by omics research, the correlation...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/89G01N1/28
CPCG01N1/30G01N30/06G01N30/89
Inventor 卢大胜汪国权周志俊薛黎明冯超金玉娥邱歆磊林元杰徐骞乐孙阳
Owner SHANGHAI MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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