High performance liquid chromatography tandem mass spectrometry detection method of 25-hydroxy vitamin D in serum

Abstract

The invention discloses a high performance liquid chromatography tandem mass spectrometry detection method of 25-hydroxy vitamin D in serum. The method comprises the following steps that (1) a sampleis pre-treated, specifically, an internal standard working solution and a sodium hydroxide solution are sequentially added into a serum sample and mixed to be uniform; (2) solid-liquid extraction treatment is conducted, specifically, the sample treated in the step (1) is loaded to an SLE 96 pore plate, and eluted with normal hexane, and then the eluant is blown to be dried by using nitrogen; (3) aderivatization reaction is conducted, a derivatization reagent 4-phenyl-1,2,4-triazoline-3,5-diketone (PTAD) solution is added to redissolve, a thermostatic reaction is conducted, then ethyl alcoholis added to terminate the reaction, and the product is blown to be dried by using nitrogen; and (4) redissolution is conducted, and analytical detection is conducted by adopting the high performance liquid chromatography tandem mass spectrometry based on a multiplexing system (MPX). The high performance liquid chromatography tandem mass spectrometry detection method is high in detection sensitivity, high in specificity and low in cost, meanwhile, the pre-treatment time and analysis time of the sample are effectively shortened, and the detection efficiency of the sample is improved.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a high-throughput, high-sensitivity, and low-cost detection method for 25-hydroxyvitamin D in serum by high performance liquid chromatography tandem mass spectrometry. Background technique [0002] Vitamin D is a steroid hormone that can regulate the body's calcium and phosphorus homeostasis, and its level in the body has an important impact on the normal physiological functions of the body. Vitamin D is converted into the metabolite 25-hydroxyvitamin D in the liver. 25-hydroxyvitamin D can reflect the level of vitamin D in the human body and is an important indicator for evaluating vitamin D. At present, the methods for determining 25-hydroxyvitamin D in serum mainly include enzyme immunoassay, competitive protein binding method, high performance liquid chromatography, etc.; these methods have the following problems and shortcomings: enzyme immunology methods cann...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve some problems existing in the existing 25-hydroxyvitamin D detection method, and to provide a high performance liquid chromatography tandem mass spectrometry detection method for 25-hydroxyvitamin D in serum, which has strong specificity and high accuracy High, high sensitivity, low cost of detection equipment, high analytical throughput, etc.

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