Detection method of 25-hydroxyvitamin D liquid chromatography tandem mass spectrometry in serum and kit

A hydroxyvitamin and liquid chromatography technology, which is applied in the field of 25-hydroxyvitamin D detection, can solve the problems of time-consuming and large differences in test results, and achieve the effects of shortened time, high accuracy, and high detection throughput

Inactive Publication Date: 2019-01-15
天津国科医疗科技发展有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the different processing methods of reagents, standards and samples used by different laboratories, it often leads to large differences in test results between different laboratories, and a lot of time is consumed in pre-processing

Method used

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  • Detection method of 25-hydroxyvitamin D liquid chromatography tandem mass spectrometry in serum and kit
  • Detection method of 25-hydroxyvitamin D liquid chromatography tandem mass spectrometry in serum and kit
  • Detection method of 25-hydroxyvitamin D liquid chromatography tandem mass spectrometry in serum and kit

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Experimental program
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Effect test

Embodiment 1

[0051] This embodiment is a liquid chromatography tandem mass spectrometry detection method for 25-hydroxyvitamin D in serum:

[0052] 1. Main experimental steps:

[0053] (1) Prepare internal standard working solution: use 25-hydroxyvitamin D provided by Toronto Research Chemicals (TRC), Canada 2 -d 6 , 25-Hydroxyvitamin D from Medical Isotopes 3 -d 6 ; Calculate the amount of the internal standard working solution according to the amount of the sample in the current experiment. Mix the above-mentioned internal standards to obtain a high-concentration internal standard mixture. When in use, dilute the high-concentration internal standard mixture with methanol and mix thoroughly to obtain an internal standard working solution.

[0054] The specific preparation method of the above-mentioned internal standard mixed solution is:

[0055] 1) According to different packaging specifications, first dilute the two internal standard stock solutions to 20μg / mL, and record them as I...

Embodiment 2

[0095] This embodiment is a kit for the detection of 25-hydroxyvitamin D in serum by liquid chromatography tandem mass spectrometry:

[0096] The kit of this implementation can detect 25(OH)D at the same time 2 and 25(OH)D 3 .

[0097] The kit of this embodiment includes: calibrator, quality control substance, internal standard mixture, precipitant, extraction agent, complex solution, mobile phase, see Table 4 for details. The internal standard mixture includes an internal standard product, and the internal standard product adopts 25-hydroxyvitamin D provided by Toronto Research Chemicals (TRC) in Canada. 2 -d 6 , 25-Hydroxyvitamin D from Medical Isotopes 3 -d 6 .

[0098] Table 4 Composition of the kit

[0099]

[0100] (1) Use of precipitant containing internal standard mixture:

[0101] Mix the internal standard in the kit into the precipitant and vortex to mix. .

[0102] (2) The formula used in the mobile phase:

[0103] The mobile phase A adopts 0.1v / v% formi...

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Abstract

The invention provides a detection method of 25-hydroxyvitamin D liquid chromatography tandem mass spectrometry in serum and a kit. The detection method of the 25-hydroxyvitamin D liquid chromatography tandem mass spectrometry in serum comprises the following steps that (1) serum samples are taken in a sample tube; (2) a precipitant containing an internal standard mixture is added into each sample, and vortexing and mixing is performed; (3) an extractant is added into each sample, and vortexing and mixing is performed; (4) supernatant is taken in the sample tube after the samples are subjectedto standing; and (5) after the samples are blown-dry by nitrogen at the room temperature, reconstitution fluid is added, after vortexing, the liquid chromatography tandem mass spectrometry is used for detection, and the content of 25-hydroxyvitamin D in serum is determined by comparing an internal peak area of the 25-hydroxyvitamin D and the 25-hydroxyvitamin D. The method for detecting the 25-hydroxyvitamin D liquid chromatography tandem mass spectrometry in serum can further improve the detection accuracy and precision and shorten the pretreatment time.

Description

technical field [0001] The invention belongs to the technical field of 25-hydroxyvitamin D detection, in particular to a method capable of simultaneously detecting 25-hydroxyvitamin D 2 and 25-hydroxyvitamin D 3 methods and kits. Background technique [0002] Vitamin D has a classic role in regulating calcium and phosphorus metabolism and a non-classical role in affecting cell proliferation and differentiation, and is an essential steroid hormone for maintaining human health. Vitamin D deficiency can lead to rickets in children and rickets in adults. In recent years, in-depth studies have shown that vitamin D deficiency increases the risk of common multiple diseases such as tumors, cardiovascular diseases, autoimmune diseases and mental diseases. Based on the importance of vitamin D to human health and the low level of vitamin D in the Chinese population, it is necessary to popularize the detection of vitamin D levels in the Chinese population and to screen key population...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 李艳许舒欣李小强胡玮贾明正程文播
Owner 天津国科医疗科技发展有限公司
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