High performance liquid chromatography tandem mass spectrometry for the detection of 25-hydroxyvitamin D in dried blood tablets

Abstract

The invention discloses a high-performance liquid chromatography tandem mass spectrometry detection method for 25-hydroxyvitamin D in dried blood slices. Internal standard working solution, vortex mixed, centrifuged, take the upper layer extract, blow dry with nitrogen; (2) derivatization reaction, step (1) processed sample, add PTAD derivatization working solution to redissolve, constant temperature reaction, then add ethanol Terminate the reaction, blow dry with nitrogen; (3) redissolve, and analyze and detect by high performance liquid chromatography tandem mass spectrometry. The method of the present invention has high efficiency, high throughput, low cost, simple operation, high accuracy, high sensitivity, no conflict limit with other detection items, and is suitable for rapid detection of large quantities of clinical samples; The trauma is relatively small, the amount of blood taken is small, the sample collection and transportation are convenient, easy to store, and the stability is good. It is especially suitable for the detection of people with difficulty in venous blood collection (such as infants and young children).

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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