Method and kit for determining very long chain fatty acids in sample by liquid chromatography tandem mass spectrometry

A very long chain fatty acid and liquid chromatography technology, which is applied in the field of liquid chromatography tandem mass spectrometry for the determination of very long chain fatty acids in samples, can solve the problem of not finding VLCFA by liquid chromatography tandem mass spectrometry, and achieves increased detection specificity and avoidance of The effect of interference, reducing sample size

Inactive Publication Date: 2019-05-10
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there is no method for detecting VLCFA by liquid

Method used

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  • Method and kit for determining very long chain fatty acids in sample by liquid chromatography tandem mass spectrometry
  • Method and kit for determining very long chain fatty acids in sample by liquid chromatography tandem mass spectrometry
  • Method and kit for determining very long chain fatty acids in sample by liquid chromatography tandem mass spectrometry

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Embodiment 1

[0041] Example 1 Method for Determination of Serum Very Long Chain Fatty Acids by Liquid Chromatography Tandem Mass Spectrometry

[0042] 1. Experimental reagents

[0043] The sample preparation solution includes: isotope internal standard solution, acetonitrile, concentrated hydrochloric acid (12mM), n-hexane, methanol (reconstituted solution).

[0044] Isotope internal standard solution: C22:0-d4 solution: concentration is 10.2ng / ml; C24:0-d4 solution: concentration is 23.2ng / ml; C26:0-d4 solution: concentration is 28.5ng / ml.

[0045] Instrument analysis fluids include:

[0046] Mobile phase A: Acetonitrile in water (v / v, 6:4) solution containing 5 mM ammonium formate.

[0047] Mobile phase B: acetonitrile isopropanol (v / v, 8:2) solution.

[0048] 2. Sample preparation

[0049] Take 50μl serum sample or very long chain fatty acid (C22:0, C24:0 and C26:0) standard solution to 10ml urinary tube, add 10μl isotope internal standard, mix and shake, add 200μl acetonitrile, add 5...

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Abstract

The invention provides a method and kit for determining very long chain fatty acids in a sample by liquid chromatography tandem mass spectrometry. The method comprises the steps that very long chain fatty acid standardized product solutions with different concentration are prepared; stable isotope internal standards are added to the standardized product solutions to be mixed uniformly, and acetonitrile and concentrated hydrochloric acid are added for sample pretreatment; then n-hexane is used for extraction, nitrogen is used for blowing dry supernatant, and liquid chromatography tandem mass spectrometry detection is performed after redissolution; a standard curve is drawn according to the concentration of the standardized product solutions and the detected signal intensity; the standardized product solutions in the step B are replaced with the sample to be detected for detection; according to a detection result of the sample to be tested and compared with the standard curve, the concentration of the very long chain fatty acids in the sample to be detected is obtained, and thus, the quantitative detection of the very long chain fatty acids in the sample is realized. The method for determining the very long chain fatty acids in the sample by liquid chromatography tandem mass spectrometry is successfully established, and the method can quickly and specifically detect the content of the very long chain fatty acids in samples (especially serum samples) with a small use amount of samples.

Description

technical field [0001] The invention relates to liquid chromatography tandem mass spectrometry detection technology, in particular to a method and a kit for liquid chromatography tandem mass spectrometry determination of very long-chain fatty acids in samples. Background technique [0002] Peroxisome diseases are inherited metabolic disorders caused by defective peroxisome biogenesis or deficiency of a single peroxisome. Peroxisome diseases are composed of many diseases, the most common of which is adrenoleukodystrophy (X-linked adrenoleukodystrophy, X-ALD). The disease shows that the oxidation of VLCFA by peroxisomes in the cells is impaired, so that VLCFA accumulates in a large amount in blood, white matter, adrenal cortex and other organs and tissues, causing demyelination of the central nervous system and adrenal cortex shrinkage or dysplasia. , mainly involving the adrenal glands and white matter. Serum very long chain fatty acid (VLCFA) is mainly composed of 22 and m...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72
Inventor 王丹晨禹松林邱玲
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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