Method for simultaneously detecting multiple water-soluble vitamins in serum based on liquid chromatography-tandem mass spectrometry

Abstract

The invention provides a method for simultaneously detecting various water-soluble vitamins in serum based on liquid chromatography-tandem mass spectrometry, which comprises the following steps: adding a proper amount of internal standard solution into a serum sample, then adding an extracting agent I, carrying out vortex oscillation to uniformly mix the sample, standing, then adding an extracting agent II into the sample, carrying out vortex oscillation, and carrying out gradient elution. And transferring all the samples into an ultrafiltration tube, carrying out high-speed centrifugation, taking out the ultrafiltration tube, transferring the centrifuged extracting solution at the lower layer into a sample feeding plate, adding an extracting agent III, carrying out vortex oscillation, uniformly mixing, and loading the sample onto a liquid chromatography-tandem mass spectrometer for detection. The method can rapidly and accurately detect 11 water-soluble vitamins in serum at the same time, pretreatment is simple and easy to operate, time and labor are saved, high efficiency and accuracy are achieved, 11 water-soluble vitamins are detected through one-time testing and one-needle sample injection, the efficiency and accuracy of medical examination are greatly improved, convenience is brought to doctors and patients, and the method is worthy of popularization and application. And an effective and feasible basis is provided for monitoring the content of the water-soluble vitamins in a human body.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a method for simultaneously detecting multiple water-soluble vitamins in serum based on liquid chromatography tandem mass spectrometry. Background technique [0002] Water-soluble vitamins are a type of organic matter necessary to maintain human life activities, participate in the regulation of body metabolism, and are also important active substances to maintain human health. Its content in the human body is very small, but it is indispensable. If a certain vitamin is lacked for a long time, it will cause physiological dysfunction and certain diseases. [0003] Thiamine (VB1) can maintain circulation, digestion, normal functions of nerves and muscles, regulate gastrointestinal function, a component of decarboxylase coenzyme, participate in glucose metabolism, and prevent beriberi; deficiency can cause neuritis, beriberi, appetite Symptoms such as malaise, indiges...

AI Technical Summary

Problems solved by technology

1. Low throughput, less target detection

2. The pretreatment purification is not thorough enough to easily contaminate the chromatographic column and mass spectrometer; 3. The sample pretreatment is complicated and the preparation cost is high

[0012] CN 111398439 A adopts the protein precipitation method to extract and purify the sample, the purification effect is very poor, and impurities such as pigment and fat are difficult to remove, which will cause relatively large pollution to the chromatographic column and mass spectrometry system, and cause multiple dilution of the sample, reducing the detection sensitivity

[0013] After searching the prior art, in the patent literature with the publication number CN 111398439 A, the types of vitamins detected are relatively small, only 6 kinds are detected, and several of them are not active substances in the human body (vitamin B6 needs to detect the active substance phosphoric acid Pyridoxal, vitamin B9 needs to detect the active substance 5-methyltetrahydrofolate, vitamin B12 needs to detect the active substance methylmalonic acid), and does not detect the difficult-to-detect pyridoxal phosphate and methylmalonic acid, no Solve multiple problems simultaneously

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