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Liquid chromatography-tandem mass spectrometry detection method of 25-hydroxyvitamin D in serum or blood plasma, and kit thereof

A technology of hydroxyvitamin and liquid chromatography, which is applied in the field of detection of 25-hydroxyvitamin D by improved liquid chromatography tandem mass spectrometry, which achieves the effects of simple pretreatment, improved detection accuracy and improved sensitivity

Inactive Publication Date: 2016-08-31
北京洛奇医学检验实验室股份有限公司
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  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve some problems existing in the existing 25-hydroxyvitamin D detection method technology, and to provide a liquid chromatography tandem mass spectrometry detection method and a kit for 25-hydroxyvitamin D in serum or plasma

Method used

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  • Liquid chromatography-tandem mass spectrometry detection method of 25-hydroxyvitamin D in serum or blood plasma, and kit thereof
  • Liquid chromatography-tandem mass spectrometry detection method of 25-hydroxyvitamin D in serum or blood plasma, and kit thereof
  • Liquid chromatography-tandem mass spectrometry detection method of 25-hydroxyvitamin D in serum or blood plasma, and kit thereof

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[0065] 1. Take a 1.5ml centrifuge tube, add 10 μl of internal standard solution and 200 μl of human serum, mix well (mix on a vortex for 45 seconds), then add 600 μl of acetone, mix on a vortex for 50 seconds, and let stand at -20°C for 20 minutes Afterwards, place it on a centrifuge at 13000rpm, centrifuge for 5min, take the supernatant into a new 1.5ml centrifuge tube, and dry it with nitrogen at 50°C. Add 200 μl of acetonitrile to redissolve, mix on a vortexer for 50 seconds, place on a centrifuge at 13,000 rpm, centrifuge for 3 minutes, and take the supernatant to obtain the final product.

[0066] 2. The detection method is liquid chromatography tandem mass spectrometry, in which multiple reaction monitoring (MRM) scanning is used.

[0067] Wherein the chromatographic conditions are as follows:

[0068] Chromatographic column: 4.6×30mm, 3.5μm

[0069] The flow rate of the initial mobile phase was 1ml / min, the column temperature was 30°C, the detection time was 6min, and...

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Abstract

The invention provides a liquid chromatography-tandem mass spectrometry detection method of 25-hydroxyvitamin D in serum or blood plasma. The method comprises the following steps: 1, preprocessing a sample: adding 10[mu]l of an internal standard solution and 200[mu]l of human serum to a centrifuge tube, uniformly mixing, adding 600[mu]l of acetone, uniformly mixing, standing the obtained solution at a low temperature for 15min, adding the solution to a centrifuge, centrifuging the solution at 13000rpm for 5min, taking the obtained supernatant to a second centrifuge tube, drying the supernatant with 50DEG C nitrogen until the supernatant is dry, adding 200[mu]l of acetonitrile to redissolve the dried supernatant, uniformly mixing, adding the obtained acetonitrile solution to the centrifuge, centrifuging the acetonitrile solution at 13000rpm for 2-4min, and taking the obtained supernatant; and 2, detecting the finally obtained supernatant through liquid chromatography-tandem mass spectrometry in a multi-reaction monitoring MRM scanning manner. A preprocessing technology is improved, so the preprocessing process in the invention is simple and fast, and bath processing can be realized.

Description

technical field [0001] The invention relates to the technical field of 25-hydroxyvitamin D detection, in particular to an improved liquid chromatography tandem mass spectrometry method for detecting 25-hydroxyvitamin D. Background technique [0002] In the past decade, a large number of studies have shown that vitamin D plays an important role in health and disease (HolickMF. Deficiency of sunlight and vitamin D. BMJ 2008; 336:1318–9. Holick MF, ChenTC. Vitamin D deficiency: a worldwide problem with health consequences. Am JClin Nutr 2008;87:1080S–6S.). Vitamin D is not only closely related to rickets, osteoporosis, etc., but also related to cancer, hypertension, diabetes, multiple sclerosis, depression, etc. (Holick MF. Vitamin D deficiency. NEngl J Med2007; 357:266–81.) . Research has linked vitamin D levels in childhood to many disorders in late childhood or adulthood (McGrath J. Does 'imprinting' with low prenatal vitamin D contribute to the risk of various adult disor...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 何健郭玉梅宋晓涛
Owner 北京洛奇医学检验实验室股份有限公司
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