Gas chromatography-mass spectrometry detection method of three very long chain fatty acids in human serum

Abstract

The invention provides a gas chromatography-mass spectrometry detection method of three very long chain fatty acids in a human serum. The method comprises the following steps (1) carrying out pretreatment on a human serum sample to obtain a derivative product; (2) detecting the derivative product by using a gas chromatography-mass spectrometer; (3) carrying out qualitative analysis on each very long chain fatty acid in the human serum sample according to a total ion chromatogram and a mass spectrogram of the derivative product, and calculating the content of each very long chain fatty acid in the human serum sample according to a selective ion chromatogram, wherein each very long chain fatty acid comprises docosanoic acid, tetracosanoic acid and hexacosanoic acid. Compared with the background art, the pretreatment process disclosed by the invention is fast, sensitive and accurate.

Description

technical field [0001] The invention relates to the technical field of very long-chain fatty acid detection, in particular to a gas chromatography-mass spectrometry detection method for three very long-chain fatty acids in human serum. Background technique [0002] The VLCFA (very-long-chain fatty acids) in the body mainly include docosanoic acid (C22:0), tetracosanoic acid (C24:0) and hexacosanoic acid (C26:0 ) 3 species, which can only undergo β-oxidation in peroxisomes. VLCFA mainly exists in the form of esters in the body, including triglycerides, phospholipids, glycolipids, etc., but a small part exists in free form or bound to serum albumin. [0003] Determination of the content of VLCFA in serum can be used to assist in the diagnosis of genetic metabolic diseases related to suspected peroxisome metabolic disorders, including peroxisome biogenic disorders, X-linked adrenoleukodystrophy and peroxidase Physical function assessment, etc. [0004] Currently, gas chromat...

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Problems solved by technology

[0009] For the removal of sterol impurities, when the silane derivatization method is used, the solution is acidified with hydrochloric acid after hydrolysis, and extracted with n-hexane. This step extracts fatty acids and non-polar sterol compounds at the same time, and then adds hydroxide Potassium, which causes the saponification reaction of fatty acids, exists in the water phase layer in an ionized state, and the sterol impurities without carboxyl groups remain in the organic phase layer because they have not undergone saponification reaction, and are removed by discarding the organic phase layer Most of the sterol impurities, and then add hydrochloric acid again in the water phase layer, so that the ionized fatty acid is in a free state, add n-hexane to extract, blow dry with nitrogen, and carry out derivatization reaction. This method has cumbersome steps, complicated operation, and high extraction efficiency Low, not conducive to obtaining stable results

However, there is no step involving the removal of sterols in the pretreatment method using the methyl esterification reaction

[0010] Since the pretreatment methods reported so far for the determination of very long chain fatty acids in serum are very complicated, it is particularly important to establish a fast, sensitive and accurate pretreatment method

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