Ultra-high performance liquid chromatography-mass spectrometry method for determining isotope dilution

Abstract

The invention discloses an ultra-high performance liquid chromatography-mass spectrometry method for determining isotope dilution. The method comprises the following steps of 1) selecting three targetcompounds: pyridoxal, 5-methyltetrahydrofolic acid, and mecobalamine, 2) establishing a correction curve by taking the concentration ratio of the target compound to the isotope internal standard thereof as an X axis and the peak area ratio of the target compound to the isotope internal standard thereof as a Y axis, and calculating the contents of the three water-soluble vitamins, 3) preparing a protein precipitant, 4) pretreating a serum sample, and 5) preparing and processing a standard curve. A serum sample is treated in a pretreatment mode of combining protein precipitation and liquid-liquid extraction, on one hand, protein denaturation sedimentation in the sample is removed, and on the other hand, lipid impurities are removed, so that the treated sample is less interfered by a matrixin the detection process; a relatively simple pretreatment mode and a rapid detection method are applied to establish a method capable of simultaneously detecting multiple water-soluble vitamin metabolites.

Description

technical field [0001] The invention relates to a liquid chromatography-mass spectrometry method for determining isotopes of various water-soluble vitamin metabolites in serum, in particular to an isotope dilution ultra-high performance liquid chromatography-mass spectrometry method for determining multiple water-soluble vitamin metabolites in serum. use method. Background technique [0002] Water-soluble vitamins are a group of vitamins that can be dissolved in water and are components of coenzymes or prosthetic groups. It mainly includes B vitamins, which play an important role in the catalysis of enzymes, and the content of their metabolites is an important basis for judging the level of human health. For example, vitamin B6 is also called pyridoxine, which plays a regulatory role in the metabolic process of protein. It helps the production of energy and makes people feel energetic. It is called a refreshing nutrient. Pyridoxal is the main metabolite of vitamin B6. It i...

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Problems solved by technology

[0005] CN 201910837920.1 Invention title "A method for detecting water-soluble vitamins in blood", using LC-MS method to detect 11 kinds of water-soluble vitamins in blood samples, although the patent analyzes more vitamins, but only the protein precipitation used The ability to remove protein and matrix is ​​poor, and the analysis time of one needle is 15 minutes, which is difficult to apply in clinical testing

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