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

Abstract

The invention discloses a high-performance liquid chromatography (HPLC)-tandem mass spectrometry detection method of 25-hydroxy vitamin D in dried blood spots. The method comprises the following steps: (1) ultrasonic extraction: taking a dried blood spots sample, adding an extraction solvent, ultrasonically extracting, and then adding an internal standard working solution, performing vortex-mixing, centrifuging, taking upper extraction liquid, drying through nitrogen; (2) derivatization reaction: adding the sample processed in the step (1) into a PTAD derivatization working solution to redissolve, reacting at constant temperature, and adding the ethanol to terminate the reaction, and drying through the nitrogen; and (3) redissolving: performing analysis detection by adopting the HPLC-tandem mass spectrometry. The method disclosed by the invention is high in efficiency, high in flux, low in cost, simple for operation, high in accuracy, and high in sensitivity, free form conflict limitation on other detection projects, and suitable for fast detecting batch of clinical samples; and meanwhile, the wound on the subject is small by adopting the dried blood spots method, the amount of collecting blood is less, the sample is convenient for collection and transportation, and easy to save, the stabiligy is good, and the method is especially suitable for the detection on the population (such as infants) difficult for venous blood collection.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a 25-25- High performance liquid chromatography tandem mass spectrometry method for the detection of hydroxyvitamin D. Background technique [0002] The content of 25-hydroxyvitamin D in infants and young children is closely related to a series of diseases such as rickets, osteoporosis, and osteomalacia. If it is not detected in time, the disease will be delayed and treatment will be delayed. However, the prior art mainly has the following two defects: [0003] (1) The existing methods need to collect serum samples from the human body, and it is difficult for infants to collect sufficient serum, so the existing methods cannot meet the needs of special populations such as infants. The dried blood spot method is convenient for collection, requires less blood, is simple to store, and is easy to transport, making it more suitable for the detection of infants and you...

AI Technical Summary

Problems solved by technology

[0004] (2) Most of the existing detection methods use the immunological method, which cannot accurately distinguish 25-hydroxyvitamin D based on the fact that the antibody can bind to the structural analogue of the analyte 2 and 25-hydroxyvitamin D 3 , so only the total 25-hydroxyvitamin D level can be detected. At the same time, the accuracy of the detection results is not high, and it cannot accurately guide clinicians to diagnose diseases and take medicines

This method has the following multiple defects: (1) in the pretreatment, derivatization and redissolution processes, a single centrifuge tube and a single sample bottle are used to process the detection sample, the sample processing process is cumbersome, the workload is large, extremely time-consuming, and uncomfortable It is suitable for the detection and analysis of a large number of clinical samples; (2) the derivatization reaction is incubated at room temperature, which takes 1 hour, which takes a long time; (3) the instrument analysis time of a single sample takes 5 minutes, and the proportion of organic solvents in the gradient program is relatively high , these factors lead to long time-consuming analysis of samples, low detection efficiency, and large consumption of organic solvent methanol; Compared with the method, the scope of use of the APCI source is narrow, and there are fewer compounds suitable for analysis, so it causes great difficulties and restrictions for the development of other detection items on the equipment, and the daily cleaning and maintenance are also more complicated.

In short, this method has high labor costs, equipment maintenance costs, and reagent costs, and its practical application range is relatively narrow. There are obvious defects and limitations in clinical promotion, and it cannot meet the multi-item and The need for large sample volume testing

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