Rapid and sensitive analysis method for 25-hydroxy vitamin D based on immune mass spectrometry technology

A hydroxyvitamin and analysis method technology, applied in the field of immuno-mass spectrometry, can solve the problems of increasing labor burden and time-consuming, and achieve the effect of accurate vitamin D metabolite quantification and improved analysis sensitivity

Pending Publication Date: 2020-11-03
SUZHOU SYM BIO LIFESCI CO LTD
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Problems solved by technology

However, this process needs to dry and derivatize the extracted sample and then dilute it on the machine, w

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  • Rapid and sensitive analysis method for 25-hydroxy vitamin D based on immune mass spectrometry technology
  • Rapid and sensitive analysis method for 25-hydroxy vitamin D based on immune mass spectrometry technology
  • Rapid and sensitive analysis method for 25-hydroxy vitamin D based on immune mass spectrometry technology

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Embodiment

[0051] A method for rapid and sensitive analysis of 25-hydroxyvitamin D based on immuno-mass spectrometry is provided, and the steps include:

[0052] 1. Prepare isotope internal standard solution:

[0053] Select the isotopic internal standard compound to prepare the internal standard solution according to the needs, such as 2 H, 13 C and 18 Compounds labeled with stable isotopes such as O can be used as internal standards for the corresponding 25-hydroxyvitamin D2 and D3 compounds to prepare internal standard solutions. if available 2 h 3 -25 hydroxyvitamin D2 and 2 h 3 -25Hydroxyvitamin D3 was used to prepare internal standard solutions so that the final concentrations reached 50 ng / mL respectively.

[0054] 2. Preparation of biotinylated antibody magnetic beads:

[0055] A1. Preparation of biotinylated antibody:

[0056] A1-1, take 1mg 25(OH)D antibody;

[0057] A1-2. Dialyze with pH 8.0 phosphate buffer at 4°C or room temperature for 16-24 hours;

[0058] A1-3....

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Abstract

The invention relates to a rapid sensitivity analysis method for 25-hydroxyvitamin D based on an immune mass spectrometry technology, which comprises the following steps: S1, enrichment and purification: performing sampling, adding an isotope internal standard solution and biotinylated antibody magnetic beads, carrying out oscillation incubation, and sequentially washing the sample with a magneticbead cleaning solution and pure water after incubation; s2, one-step derivative elution: adding the washed magnetic beads into an acetonitrile solution containing 4-phenyl 1, 2, 4-triazoline 3, diketone, carrying out oscillation incubation to carry out derivative elution, and adding an aqueous solution containing CH3NH2 into a derivative eluent to terminate the reaction; and S3, liquid chromatography-mass spectrometry detection: transferring the derivative eluent to a liquid chromatography-mass spectrometry instrument for detection. According to the method, a sample one-step method derivativeelution strategy is creatively invented in the elution stage of the vitamin D metabolite of the sample, so that the analysis sensitivity of the immune mass spectrometry is improved, the accurate quantification of the vitamin D metabolite is realized, and the additional sample treatment time and steps are not increased.

Description

technical field [0001] The invention relates to the technical field of immune mass spectrometry, in particular to a rapid and sensitive analysis method for 25-hydroxyvitamin D based on immune mass spectrometry technology. Background technique [0002] 25 hydroxyvitamin D, or 25(OH)D, includes 25 hydroxyvitamin D2, or 25(OH)D2 and 25hydroxyvitamin D3, or 25(OH)D3, which is the main form of vitamin D in the body. The detection methods of 25(OH)D2 and 25(OH)D3 in biological samples (such as whole blood, serum, urine or saliva) mainly include liquid chromatography-mass spectrometry and immunoassay. [0003] Before detection by liquid chromatography mass spectrometry, 25(OH)D2 and 25(OH)D3 need to be enriched and separated from biological samples. Traditional sample processing methods mainly include precipitation, liquid-liquid extraction and solid-phase extraction. The substances in the obtained samples are separated by liquid chromatography according to their different propert...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/08G01N30/14G01N30/72G01N33/82G01N33/53
CPCG01N30/02G01N30/06G01N30/08G01N30/14G01N30/72G01N33/82G01N33/53G01N2030/067
Inventor 姜宽祝长城许海峰
Owner SUZHOU SYM BIO LIFESCI CO LTD
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