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A method for qualitative and quantitative analysis of free fatty acids based on double derivatization technology

A free fatty acid, double derivatization technology, applied in the direction of analyzing materials, material analysis by electromagnetic means, measuring devices, etc., can solve the problems of insufficient detection sensitivity, difficult identification of double bond positions, low ionization efficiency, etc. , high sensitivity, high mass spectral response effect

Active Publication Date: 2021-07-27
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The technical problem to be solved by the present invention is to provide a qualitative and quantitative analysis method of free fatty acid mass spectrometry based on double derivatization technology in view of the above-mentioned deficiencies in the prior art, which solves the difficulty in identifying the double bond position of unsaturated fatty acids, especially polyunsaturated fatty acids , the difficulties of low ionization efficiency and insufficient detection sensitivity in the negative ion mode have achieved high sensitivity and accurate qualitative analysis

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  • A method for qualitative and quantitative analysis of free fatty acids based on double derivatization technology
  • A method for qualitative and quantitative analysis of free fatty acids based on double derivatization technology
  • A method for qualitative and quantitative analysis of free fatty acids based on double derivatization technology

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Embodiment 1

[0047] Qualitative and Quantitative Analysis of Free Fatty Acids in Example 1 Human Gastric Cancer Cells

[0048] A method for qualitative and quantitative analysis of free fatty acids in human gastric cancer cells based on double derivatization technology, the specific steps are as follows:

[0049] 1) Take one million cells into a 16mm×125mm glass tube, add 150μL of 10μmol / L C17:1(n-7) and 1ml PBS, centrifuge for 2 minutes at 3000 rpm, and discard the upper layer; Then add 1mL DPBS, then add 2mL methanol, acidify with hydrochloric acid, the final concentration of hydrochloric acid reaches 25mM; then add 1ml isooctane, centrifuge at 3000 rpm for 1 minute, separate layers, and move the upper layer to a 10mm×75mm glass tube medium, blow dry with nitrogen; after the fatty acid extract was reconstituted with 1ml acetone:water (6:4), take 50μL dilute liquid nitrogen and blow dry, finally reconstitute with 950μL acetone:water (6:4), add 50μL ethanol, and use In the subsequent phot...

Embodiment 2

[0052] Qualitative and quantitative analysis of free fatty acids in embodiment 2 human plasma

[0053] Qualitative and quantitative analysis of free fatty acids in human plasma: Take 10 μL into a 16mm×125mm glass tube, add 150 μL of 10 μmol / L C17:1(n-7), add 1ml of PBS, centrifuge for 2 minutes, 3000 rpm, and then Discard the supernatant. Add 1 mL of DPBS to the cells, followed by 2 mL of methanol, and acidify with hydrochloric acid to a final concentration of 25 mM. After adding 1ml of isooctane, the sample was centrifuged at 3000 rpm for 1 minute, separated into layers, and the upper layer was transferred to a 10mm×75mm glass tube, blown dry with nitrogen, and used for derivatization reaction. After the fatty acid extract was reconstituted with 1ml of acetone:water (6:4), take 50 μL of dilute liquid nitrogen to blow dry, reconstitute with 950 μL of acetone:water (6:4), add 50 μL of ethanol for the subsequent photochemical derivatization reaction and N, N-diethylethylenedia...

Embodiment 3

[0056] Qualitative and quantitative analysis of free fatty acids in the mouse liver in embodiment 3

[0057] Qualitative and quantitative analysis of free fatty acids in mouse liver: Take 1 mg into a 16mm×125mm glass tube, add 100 μL of 40 μmol / L C17:1(n-7), add 1ml of PBS, centrifuge for 2 minutes, 3000 rpm, Then discard the upper aqueous layer. Add 1 mL of DPBS to the cells, followed by 2 mL of methanol, and acidify with hydrochloric acid to a final concentration of 25 mM. After adding 1ml of isooctane, the sample was centrifuged at 3000 rpm for 1 minute, separated into layers, and the upper layer was transferred to a 10mm×75mm glass tube, blown dry with nitrogen, and used for derivatization reaction. After the fatty acid extract was reconstituted with 1ml of acetone:water (6:4), take 50 μL of dilute liquid nitrogen to blow dry, reconstitute with 950 μL of acetone:water (6:4), add 50 μL of ethanol for the subsequent photochemical derivatization reaction and N, N-diethyleth...

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Abstract

The invention relates to a qualitative and quantitative analysis method of free fatty acid mass spectrometry based on double derivatization technology, comprising the following steps: 1) photochemical derivatization reaction of unsaturated fatty acid double bond; amine derivatization reaction; 3) qualitative method for the precise structure of fatty acid; 4) quantitative method for fatty acid. The present invention uses the established method to carry out accurate qualitative and quantitative analysis of free fatty acids in actual samples, which solves the difficulty of identifying the double bond position of unsaturated fatty acids, especially polyunsaturated fatty acids, and the low ionization efficiency and detection sensitivity of fatty acids in negative ion mode Insufficient difficulties have achieved high sensitivity and accurate qualitative and quantitative analysis of free fatty acids in actual samples.

Description

technical field [0001] The invention relates to a qualitative and quantitative analysis method of free fatty acid mass spectrometry based on double derivatization technology, and its application in the qualitative and quantitative analysis of free fatty acids in different biological samples. [0002] technical background [0003] Fatty acid (FA) refers to a series of organic substances with long aliphatic hydrocarbon chains containing a carboxyl group at one end. The length of the carbon chain varies (C4-C36). It is divided into saturated and unsaturated fatty acids according to whether they contain double bonds. FA is the simplest class of lipid compounds and is an important component of other lipids. The diversity of fatty acids in mammals plays a variety of key roles in cellular functions through: for example, the composition of various functional lipid molecules in biological membranes; the regulation of membrane fluidity and the interaction between signaling molecules; ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/62G01N30/02G01N30/06G01N30/72G01N1/28
CPCG01N1/28G01N27/62
Inventor 魏芳徐淑玲吕昕董绪燕陈洪黄凤洪
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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