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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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  • Abstract
  • Description
  • Claims
  • Application Information

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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, which will introduce additional operation steps, which is very time-consuming and increases the labor burden

Method used

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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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Experimental program
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Effect test

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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