Method for detecting 25(hydroxyl)vitamin D by using high-pass liquid chromatography-tandem mass spectrometry

Abstract

The embodiment of the invention provides a method for detecting 25(hydroxyl)vitamin D by using a high-pass liquid chromatography-tandem mass spectrometry. The method comprises the following steps of: adding an acetonitrile solution containing an internal standard substance of the 25(hydroxyl)vitamin D into a human serum sample to carry out protein precipitation; sufficiently and uniformly mixing the solution, and then, adding an n-hexane extracting solvent; sufficiently and uniformly mixing the solution, then centrifuging the solution, movably taking a supernatant and drying the supernatant, and adding a complex solution to obtain a sample to be detected; detecting the sample to be detected by using a high-pass liquid chromatography-tandem quadrupole mass spectrometer; and quantifying according to the relative retention time of 25(hydroxyl)vitamin D2 and / or 25(hydroxyl)vitamin D3 and the detected abundance ratio of quantitative ion pairs by using an internal standard curve method. According to the embodiment of the invention, the method has the advantages of simplicity in pretreatment, strong specificity and matrix interference resistance, short detection time, high pass, high detection precision and low cost.

Description

technical field [0001] The invention relates to the technical field of vitamin D detection, in particular to a method for detecting 25-hydroxyvitamin D by high-throughput liquid chromatography tandem mass spectrometry. Background technique [0002] Vitamin D is a sterol derivative, and 25-hydroxyvitamin D is the product of vitamin D oxidized by hepatic monooxygenase-25 hydroxyloxidase. Compared with other metabolites of vitamin D, its concentration in blood is high and The cycle is long, and it becomes a good indicator of the concentration of vitamin D in the body, and it is an evaluation index of the nutritional status of vitamin D in the human body. [0003] The most important members of the vitamin D family are vitamin D2 and vitamin D3. Vitamin D2 is mostly contained in plant foods, and it is synthesized by plant ergosterol through sunlight. Vitamin D3 can be synthesized by the human skin itself, and can also be ingested through food. Vitamin D plays an important phys...

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Problems solved by technology

[0005] Traditional 25-hydroxyvitamin D detection methods include radioimmunoassay, competitive protein binding method, high-performance liquid chromatography, etc., but there are mainly the following problems: First, because 25-hydroxyvitamin D is mainly combined with serum-binding protein DBP in the human body, Moreover, there are a large number of high-affinity binding proteins in the human body, so there is serious matrix interference in the detection

However, the traditional radioimmunoassay and competitive protein binding method cannot effectively eliminate matrix interference; second, the existing detection technology has complicated pretreatment, long analysis time, and poor method specificity; third, it cannot accurately quantify 25-hydroxyvitamin D at the same time 2 and 25 hydroxyvitamin D 3 The content of 25-hydroxyvitamin D cannot be given accurately; Fourth, the entire detection process takes a long time and the throughput is low

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